bluuebunny/crossref_metadata_2025 huggingface dataset:
Vegetarian dietary pattern is associated with increased consumption of fiber, calcium and potassium in adult respondents of the National Health and Nutrition Examination Survey, 2003–2008
Authors: Devareddy Latha , Miller Kevin , Liska DeAnn , Fulgoni Victor
Published: June 2021

Vegetarian diets have been associated with several health benefits. However, there are some concerns around the nutritional adequacy of vegetarian diets. The present study was designed to evaluate the differences in nutrient intakes between vegetarian (VDP) and non‐vegetarian dietary patterns (NVDP) in US adult population (> 19 yrs; N = 11,888) using the data from the National Health and Nutrition Examination Survey, 2003–2008. Respondents who did not report the consumption of meat, poultry or fish on the NHANES dietary recall for either of the two survey days were categorized as having a vegetarian dietary pattern. Mean nutrient intakes were adjusted for Age, Gender, Ethnicity, Poverty Income Ratio and kCal. Total energy intake was approximately. 16% lower in VDP when compared to NVDP. Mean intakes of fiber, vitamins A, C, and E, total folate, calcium, magnesium, and potassium were higher for VDP than for NVDP. Although intake of sodium was lower in the VDP, it was still higher than the current guideline. Dairy and fruit consumption was higher among the vegetarians. Contrary to the common belief, total vegetable intake was not different between the two groups. The Healthy Eating Index, a measure of diet quality, was not different for people following a vegetarian dietary pattern compared to non‐vegetarians.
Contrast ultrasound imaging does not affect Hsp70 expression in cholesterol‐fed rabbit aorta
Authors: Smith Brendon W. , Simpson Douglas G. , Miller Rita J. , Lee Matthew B. , O'Brien William D. , Erdman John W.
Published: June 2021

Diagnostic ultrasound imaging (US) is enhanced by the use of circulating microbubble contrast agents (UCAs), but the interactions between US, UCAs, and vascular tissue are not well understood. We hypothesized that US with UCA would represent a stress to the vascular tissue at the site of exposure and increase levels of Hsp70, a cellular stress protein. Male New Zealand White rabbits (n=24) were fed a diet containing 1% cholesterol, 10% fat and 0.11% magnesium. At 21 days, rabbits were either exposed to US (2.1 MPa) using the UCA Definity or sham exposed using a saline control (n=12 per group). Blood plasma was analyzed for cholesterol and von Willebrand Factor (vWF), a marker of endothelial function. Animals were euthanized 24 hours after exposure and a section of the abdominal aorta was quickly isolated and snap‐frozen in liquid nitrogen. Aorta lysates from the specific area of US exposure were analyzed for Hsp70 protein expression by Western blot. Plasma total cholesterol levels increased to an average of 705 mg/dL. Hsp70 was expressed in aortas of both groups. No significant differences were observed between the two groups in Hsp70 expression or plasma vWF levels. Elevation of plasma cholesterol is a substantial stress to the vasculature, and the stress protein Hsp70 was expressed in vascular tissue. No additional vascular stress was observed as a result of US imaging. [Supported by NIH R37EB002641]
Use of MetaboAnalyst as a tool to study metabolomics data for a dietary intervention study
Authors: Panjeton Geoffrey D. , Remz Matthew A. , Allen Hannah J. , Powell David H. , Borum Peggy R.
Published: June 2021

Background

Metabolomics is the comprehensive analysis of metabolites (<1500 Da). MetaboAnalyst (MA) is web‐based, publicly available software for analysis of quantitative metabolomics data.

Objective

To demonstrate the use of MA in a pilot study evaluating the effect of diet change on the human plasma metabolome.

Methods

Plasma collected from healthy adult females before and after 5 days of receiving a high‐fat, low‐carbohydrate, and adequate protein diet were analyzed using HPLC‐ESI‐TOF‐MS. Features (mass/retention time) and abundances were uploaded to MA for analysis. PCA and PLS‐DA were used to differentiate between groups. Features were selected based on significance, using a volcano plot (p of 0.1 and fold change of 2). The remaining features were filtered based on presence in samples. A heatmap was used to identify clusters of features.

Results

The volcano plot showed that less than 5% of the features were significant. The number of features was reduced from 2225 to 81 and 48 of these features were present in 50% of the subjects. Using these targeted 48 features, the PCA, PLS‐DA, and heatmap showed improved clustering of subjects based on diet group compared to the global analysis.

Conclusions

MA can be used to evaluate the effects of a diet change on the plasma metabolome.

Supported in part by the National Institutes of Health (NIH) and National Center for Research Resources (NCRR) CTSA grant 1UL 1RR029890.
Comparison of Serum Levels of Lipids and Inflammatory Markers in Korean Young Adult Men by Fat Intakes
Authors: Kim soon kyung , Lee sung hye , Choi mi kyeong
Published: June 2021

The objective of this study was to investigate the impact of fat consumption of men in 20's on their lipids levels, obesity‐related hormones, and inflammatory markers. The subjects were classified into two groups according to fat intake levels to compare their nutrient intake, physical measurements, lipids levels, obesity‐related hormones, and inflammatory markers (hs‐TNF‐α, CRP, adiponectin). Of the 48 subjects, the number of Group I (below the average) was 27 (56%) and that of Group II (the average or above) was 21 (44%) while there was no significant difference in the physical measurements of the two groups. As to the nutrient intake, Group II showed higher cholesterol (p<0.01), total fatty acids (p<0.01), saturated fatty acids (p<0.01) and mono‐unsaturated fatty acids (p<0.01) levels. Based on food groups, Group II showed higher consumption of sugars and sugar‐based products (p<0.01), mushrooms (p<0.01), meat and meat‐related products (p<0.01), eggs (p<0.01), and seasonings/spice (p<0.05). Regarding the lipid levels, Group I had significantly higher HDL‐cholesterol (p<0.01) levels while Group II had higher triglyceride, LDL‐cholesterol, atherogenic indices, cardiac disease risk indices, and LPH. Concerning adiponectin, an obesity‐related hormone, and inflammatory markers, Group I showed higher adiponectin and TNF‐α, but lower CRP compared with Group I. In the correlation between nutrient intake, physical measurements, lipid levels, adiponectin, and inflammatory markers, it was observed that weight, BMI, and body fat was significantly related with nutrient intake and lipid levels but not with adiponectic and inflammatory markers.
Increasing the number of masticatory cycles reduces food intake in healthy young adults
Authors: ZHU YONG , HOLLIS JAMES H
Published: June 2021

To investigate if the number of masticatory cycles made during ingestion affects satiation, 47 healthy participants (16 lean, 16 overweight and 15 obese) were recruited for this present study. Pizza rolls were used as the test food and their habitual mastication parameters were measured before they were randomized to a treatment order. Participants were asked to consume the same breakfast and reported to the laboratory for lunch, once a week for three weeks. After baseline appetite measures were made they were asked to eat until they were comfortably full, by chewing each portion of the food at their habitual number of chews, 150% or 200% of their habitual number of chews. Subjective appetite was measured regularly for 60 minutes and upon meal termination. There was no effect of body mass index (BMI) on habitual number of chews, chewing rate and chewing duration. Adjusting for BMI, increased number of masticatory cycles resulted in increased meal duration (P<0.0001) and reduced food intake (P=0.0026). 200% of the habitual number of chews reduced the food intake by 15% (P=0.0007) whereas 150% of the habitual number of chews reduced the food intake by 10% (P=0.0257). The number of chews did not affect hunger, fullness, desire to eat and palatability of the test food. The results suggest increasing the number of chews promotes satiation and could be a strategy for body weight management.

Grant Funding Source : Iowa State University
Effect of zinc deficiency and diet restriction on liver and spleen non‐heme iron concentrations in rats
Authors: Konomi Aki , Yokoi Katsuhiko
Published: June 2021

Iron preparation, an important therapeutics for nutritional anemia, is not effective for some subsets of nutritional anemia, i.e., iron refractory anemia. Some researchers reported that oral zinc effectively cured some patients with iron refractory anemia. It is not established that single zinc deficiency induces anemia in humans. We reported before that Zn deficiency depressed plasma erythropoietin (EPO) concentration and kidney EPO concentration in rats. If erythropoiesis is impaired by zinc deficiency, unutilized non‐heme iron (ferritin and hemosiderin) may be accumulated in liver of zinc‐deficient rats. To test the hypothesis, thirty 3‐week‐old male Sprague‐Dawley rats were assigned into 3 groups of 10 for the 4‐week study. Difference between groups was statistically analyzed by Fisher's PLSD. P value less than 0.05 was considered significant. Liver hemosiderin concentration of zinc‐deficient group was marginally increased compared with control group (P=0.052), and significantly increased compared with pair‐fed group (P=0.031). Liver ferritin concentration of zinc‐deficient group was significantly increased compared with control group and pair‐fed group (P<0.0001 each). Spleen non‐heme iron concentrations were not changed among all treatment groups. These results suggest that hepatic iron store was increased by zinc deficiency per se, but not changed by diet restriction. This work was supported by KAKENHI grant‐in‐aid for Research Activity Start‐up (21800073) and Young Scientists (B) (23700928).
Effect of beef consumption on body iron and cognitive function in iron‐deficient women
Authors: Blanton Cynthia
Published: June 2021

Iron depletion is associated with cognitive impairment in women of reproductive age. This project investigates the effect of moderate beef intake on total body iron and cognitive function in university undergraduate women. At baseline, women (n=17) were classified by body iron tertile and randomly assigned to a beef or nonbeef meat lunch group (85 g, 3x weekly) for 16 weeks. Computerized neurocognitive tests were administered at baseline and week 16. Changes in calculated body iron and cognitive performance from baseline to endpoint were analyzed using generalized linear models and Spearman correlations. Women in the 1st (lowest) vs. 2nd and 3rd iron tertile responded to either lunch intervention with significant increases in body iron (ANOVA, P<0.01) and significant improvements in tests of spatial working memory (decrease in errors and improvement in strategy, P<0.05). Change in body iron was inversely associated with change in spatial working memory error rate (r=−0.49; P<0.05). These preliminary findings suggest that working memory improves in association with increased iron status in iron‐deficient women consuming moderate amounts of beef or nonbeef meat. Funded by the Idaho Beef Council.
Role of clathrin‐mediated endocytosis in transferrin‐bound iron uptake by hepatocytes
Authors: Zhang Lin , Knutson Mitchell
Published: June 2021

After erythroid cells of the bone marrow, hepatocytes of the liver represent the second largest consumer of transferrin‐bound iron, accounting for 10–20% of iron exchange with the plasma. Although it is well known that transferrin is taken up by cells via endocytosis, a previous study in hepatocytes found that chemical inhibitors of endocytosis did not inhibit the assimilation of iron from transferrin. The aim of the present study was to investigate the role of clathrin‐mediated endocytosis in the uptake of transferrin‐bound iron by HepG2 cells, a human hepatoma cell line. We used siRNA to suppress the expression of clathrin and then incubated the cells with 59Fe‐labeled transferrin for 1 hour. Although suppression of clathrin resulted in a 70% lower uptake of transferrin, the cellular assimilation of 59Fe iron was unaffected. These data suggest that HepG2 cells take up transferrin primarily through clathrin‐mediated endocytosis, but that endocytosis is not required for the uptake of iron from transferrin. Funded by NIH.

Grant Funding Source : NIH
Role of the iron‐import protein DMT1 (Divalent Metal Transporter 1) in liver iron uptake
Authors: Wang Chia-Yu , Knutson Mitchell
Published: June 2021

DMT1 is required for intestinal iron absorption and the uptake of iron by developing erythroid cells. Here we used hepatocyte‐specific DMT1 knockout mice (Dmt1liv/liv) to examine the role of DMT1 in the liver. Dmt1liv/liv mice were crossed with models of genetic iron overload, i.e., Hfe−/− and hypotransferrinemic (Hpx) mice, to determine the role of DMT1 in hepatic iron accumulation. We found that Dmt1liv/liv mice had normal hepatic non‐heme iron concentrations and that the double‐mutant Dmt1liv/liv:Hfe−/− and Dmt1liv/liv:Hpx mice accumulated similar amounts of hepatic iron as did their respective Hfe−/− and Hpx controls. To determine if hepatocyte DMT1 is required for the uptake of non‐transferrrin‐bound iron (NTBI) and the assimilation of iron from transferrin, we injected 59Fe‐labeled NTBI or transferrin into Dmt1liv/liv mice and measured hepatic 59Fe uptake. We found that Dmt1liv/liv mice efficiently took up NTBI, but had a 40% reduction in the assimilation of iron from transferrin. These data indicate that DMT1 is not required for hepatic iron accumulation under normal and iron overload conditions, but it is required for the efficient assimilation of iron from transferrin. Funded by NIH.

Grant Funding Source: NIH
Posttranslational Regulation of the Menkes Copper ATPase (Atp7a) in Intestinal Epithelial Cells
Authors: Xie Liwei , Collins James F.
Published: June 2021

Atp7a pumps copper into the trans‐Golgi for cuproenzyme synthesis and also functions to export copper from enterocytes. It was noted that copper treatment of rat intestinal epithelial (IEC‐6) cells increased Atp7a protein abundance without effecting mRNA levels. We thus hypothesized that intracellular copper stabilizes the Atp7a protein and studies were designed to test this possibility. Differentiated IEC‐6 cells were treated with a protein translation inhibitor (cyclohexamide [CHX]) at different concentrations for different times to estimate Atp7a protein half‐life. CHX at 10 μg/ml for 48 hours led to >50% reduction in immunoreactive Atp7a protein. Next, IEC‐6 cells were exposed to 10 μg/ml CHX in the presence or absence of added copper (150 μM) for 48 hours and Atp7a protein abundance was determined. Results showed that copper stabilized the Atp7a protein, as protein abundance in CHX/Cu treated cells was similar to control cells treated with vehicle while CHX alone led to >50% protein reduction. To determine the mechanism, further studies will be designed to assess the role of the 6 metal binding domains of Atp7a, as not all of them are required for Atp7a copper transport function. We conclude that Atp7a is stabilized by copper, which could be particularly relevant during iron deficiency when the copper content of the intestinal mucosa increases and Atp7a protein abundance in enterocytes increases dramatically.

Grant Funding Source : 1R01 DK074867
Poor vitamin D status is associated with increased risk for metabolic syndrome (MeS), diabetes mellitus (DM), and several unfavorable cardiometabolic markers in US adults
Authors: Ganji Vijay , Zhang Xu , Tangpricha Vin
Published: June 2021

We investigated the relation between serum 25 hydroxyvitamin D [25(OH)D] concentration and prevalence of MeS and DM and concentrations of cardiometabolic biomarkers in >19 y old persons using recently‐released data from NHANES, 2001–2006 (n=8241). The prevalence of MeS and DM were significantly higher in the serum 25(OH)D <30 nmol/L group (MeS, 45.5%; DM, 21.6%) compared to the serum 25(OH)D ≥75 nmol/L group (MeS, 11.6%; DM, 4.1%) (P<.001). The likelihood of having MeS (adjusted OR, 2.89; 95%CI, 2.07, 4.03) and DM (adjusted OR, 1.69; 95%CI, 1.18, 2.42) were significantly higher in the serum 25(OH)D <30 nmol/L group compared to the serum 25(OH)D ≥75 nmol/L group (P<.001). In multivariate‐adjusted analysis, the group with serum 25(OH)D <30 nmol/L had significantly higher waist circumference, glycohemoglobin (HbA1), triglycerides, insulin, C‐peptide, C‐reactive protein, and HOMA‐IR and had significantly lower VO2max compared to the group with serum 25(OH)D ≥75 nmol/L (P<.001). We conclude that persons with vitamin D deficiency are significantly higher risk for MeS and DM compared to those with vitamin D sufficiency. Vitamin D deficiency is related to several cardiometabolic risk factors. Given the association between vitamin D deficiency and poor health outcomes in these most recent NHANESs and past NHANES, studies focusing on correction of vitamin D deficiency in reducing cardiometabolic risk are needed.
Excessive Variability in a Popular Direct Immunoassay for Total 25 Hydroxy Vitamin D (t25OHD)
Authors: Holmes Earle W , Garbincius Jean , McKenna Kathleen M.
Published: June 2021

This study was undertaken to compare t25OHD results determined by the DiaSorin Liaison direct immunoassay(IA) (DiaSorin Inc., Stillwater, MN 55082) with results that were measured by LC – MS (Quest Diagnostics, Wood Dale, IL 60191).166 consecutive clinical specimens were assayed for t25OHD by both IA and LC‐MS. The paired results were analyzed by linear regression analysis. Cluster analysis was used to classify the specimens into 5 groups based on the magnitudes of the biases in the t25OHD results determined by the two methods. The mean bias in t25OHD for each group was compared to each group's mean concentrations of 25 OHD2 and 25 OHDD3. Regression parameters for the paired results measured by IA(y) and LC‐MS(x) were: y=0.85x+5.9; r2=0.64; SE = 9.1 ng/mL. The inter‐method biases between the t25OHD results by IA and LC‐MS ranged from +30 ng/mL to −30 ng/mL Evaluation of the 5 groups derived from cluster analysis showed that the two groups with the highest positive mean inter‐method biases had significantly increased 25OHD2 concentrations and significantly increased 25OHD2/25OHD3 ratios compared to the other 3 groups. t25OHD was under‐recovered by the IA in about half of the clinical specimens with 25OHD2 concentrations that were > 20 ng/mL. The inter‐method bias between IA and LC‐MS results exceeded the total allowable error specification for 25OHD measurements in 28 % of the clinical specimens that were tested. The results showed that t25OHD concentrations determined by the Liaison IA can be falsely elevated or falsely decreased relative to LC‐MS determined values by heterophile antibody interference and under‐recovery of 25OHD2, respectively. These analytical limitations emphasize the need to carefully validate the accuracy of direct immunoassays for total 25OHD against a definitive reference method before they are approved for routine clinical use.
Development and validation of a urinary biomarker for dietary furanocoumarins
Authors: Simpkins Scott William , Young Lindsay Rae , Peterson Sabrina
Published: June 2021

Furanocoumarins from food sources (e.g., parsnip, parsley, citrus) have been shown to inhibit carcinogen‐activating cytochrome P450 enzymes and may therefore play a role in prevention of cancer. Development of a biomarker to assess exposure to dietary furanocoumarins could be an alternative to recall‐based dietary assessment methods and be useful for epidemiologic studies of diet and cancer.

In a combined dose‐response and pharmacokinetics study, timed urine samples were collected from 10 men and women over 168 hours following low (2g/kg body weight) and high (4g/kg body weight) doses of furanocoumarin‐containing vegetables (parsnips and parsley). These samples were analyzed to confirm 8‐ methoxypsoralen (8‐MOP) and 5‐methoxypsoralen (5‐MOP) as suitable biomarkers of dietary furanocoumarin intake. Pooled control urine, obtained from subjects following a week‐long abstention from furanocoumarin‐containing foods, was utilized for method development.

Following β‐glucuronidase/sulfatase hydrolysis, urine samples were processed via solid phase extraction and analyzed by liquid chromatography‐mass spectrometry. Recoveries of 8‐MOP and 5‐ MOP were > 80%. Intraday coefficients of variation were < 4% for both analytes with estimated detection limits of 6.0 pM (1.3 pg/mL). With analysis times under five minutes, this method is rapid and robust enough for application to future epidemiological studies.
Fatty acid composition of erythrocytes reflects CLA intake in a depletion‐repletion study with healthy subjects
Authors: Nunes Juliana Côrtes , Penedo Letícia Abel , Torres Alexandre Guedes
Published: June 2021

We studied fatty acid (FA) composition of erythrocytes in response to CLA depletion (8 w) and CLA repletion (8 w), in 27 men and women, aged 23 to 36 years. During the depletion phase all participants excluded sources of dairy fat from their diets and in the repletion phase they returned to their habitual diets and received 20 g of CLA‐biofortified butter, resulting in the intake of 1 g CLA/d. The FA composition of erythrocytes was determined by GC after lipid extraction and FA transesterification. Twenty‐seven FA were detected in the erythrocytes. The CLA contents (g/100g FA) in erythrocytes decreased significantly (p<0.01) in the depletion phase (0.18±0.03) when compared to baseline (0.31±0.05) and increased in the repletion phase (0.34±0.06). In addition, the CLA contents in erythrocytes were positively correlated with the habitual intake of CLA food sources obtained from a food‐frequency questionnaire. The ratio between 20:4n‐6 and 20:3 n‐6 was used as indicator of Δ5‐desaturase activity and reduced (p<0.05) in the CLA repletion phase. In addition, arachidonic acid was negatively correlated (r= 0.49; p<0.05) with CLA in erythrocytes. Our results are suggestive of erythrocyte composition as marker of CLA intake and of effects of CLA on Δ5‐desaturase activity, which could mediate some of the observed biological effects of CLA.

Grant Funding Source: FAPERJ, CAPES, CNPq (Brazil).
The effects of almonds on the profile and bioaccessibility of isoflavones (IFN) from a soy soft pretzel
Authors: Simmons Amber L , Chitchumroonchokchai Chureeporn , Failla Mark L , Vodovotz Yael
Published: June 2021

Soy IFN have many purported health benefits including reducing the risk of hormone‐related cancers. β‐glucosidase activity is inherently present in almonds and converts IFN to aglycones, a potentially more bioaccessible form. To test the effects of β‐glucosidase on the bioaccessibility of IFN, a soy soft pretzel was developed as a model snack food delivery system with and without almond. Also, the lipid component was altered to assess matrix effects. Each pretzel was homogenized and subjected to a 3‐stage in vitro digestion model. IFN that partitioned in the aqueous fraction of chyme were considered bioaccessible. Each pretzel contained 95.7±8.2 μmol of total IFN. Almond added at 5.8 and 12% increased the proportion of IFN aglycones by 8.8±2.4% and 23.3±3.9%, respectively, compared to the control (2.9 shortening, p<0.05). An average of 85.2±8.0% of IFN in the starting material was recovered in the aqueous fraction regardless of the type or amount of lipid. In conclusion, almond promoted partial conversion of IFN to aglycones without affecting bioaccessibility. This implies that the health promoting potential of snack foods containing soy may be enhanced when fortified with almond.

Grant Funding Source : Ohio Agricultural Research and Development Center
Amino acids suppress the autophagic degradation pathway in skeletal muscle of septic neonatal pigs
Authors: Suryawan Agus , Orellana Renán A. , Nguyen Hanh V. , Almonaci Rosemarie D. , Davis Teresa A.
Published: June 2021

The role of amino acids (AA) in the regulation of protein degradation in skeletal muscle (SkM) of neonates is not well understood. The ubiquitin‐proteosome (UP) and autophagy‐lysosome (AL) systems are crucial proteolytic pathways that regulate SkM mass. We hypothesize that AA ameliorate the increase in autophagy induced by Lipopolysaccharide (LPS) in SkM of neonatal pigs. Overnight fasted piglets were infused with endotoxin (LPS, 0 and 10 μg/kg/hr) for 8 hr. Glucose and insulin were maintained at fasting (F) levels, AA were clamped either at F or fed levels, and degradation control mechanisms were examined in SkM. Higher AA infusion rates were required to maintain AA at F or fed levels in the LPS group when compared to controls. The abundance of atrogin‐1 and MuRF‐1 (markers of UP) and LC3‐II (marker of AL) were determined. In normal pigs, raising AA from F to fed levels had no effect on the abundance of atrogin‐1, MuRF‐1, and LC3‐II. LPS induced an increase in the abundance of atrogin‐1 and MuRF‐1 (P<0.05), but not LC3‐II. Raising AA to fed levels did not affect the sepsis‐induced changes in atrogin‐1 and MuRF‐1 abundance. However, raising AA to fed levels reduced LC3‐II abundance in septic pigs (P<0.01). In conclusion, AA have no effect on the activation of UP and AL systems under normal conditions, but in the catabolic state, AA have a robust anti‐autophagic effect. (Supported by NIH AR444474 and USDA/ARS 6250‐51000‐055)
Serum inosine, and symmetrical di‐methyl arginine (SDMA) concentrations were reduced at different time points when dogs were fed a controlled protein and phosphorus food with different fatty acid compositions
Authors: Jewell Dennis Edward , Yerramilli Maha , Obare Edward , Yerramilli Murthy
Published: June 2021

Dogs (43, 3.1 to 14.8 years of age) were assigned to one of three dietary treatment foods which was fed throughout the experiment in order to evaluate the effect of food on specific circulating metabolites. Dogs were analyzed for body composition and serum analyzed at 0, 1, 3 and 6 months. Serum analysis included urea, albumin, fatty acids, inosine and SDMA. The experimental protocol was reviewed and approved by the Institutional Animal Care and Use Committee. All foods contained approximately 14% protein and 18% fat. Fatty acid composition was as follows:

Food
12:0
18:2  n6
18:3  n3
20:5  n3
22:6  n3

Control (C)
0.01
3.07
1.27
0.01
0.0

C + fish oil (CFO)
0.01
3.09
1.35
0.10
0.07

CFO + coconut oil (CFOCO)
0.87
4.61
1.63
0.23
0.17

Serum fatty acid concentration increased in response to increased intake throughout the study. There was no change in albumin concentration or lean body mass. There was a decrease in serum urea nitrogen and inosine after one month and through the rest of the study (P<0.05). There was no change in SDMA at one month but a significant decrease (P<0.05) through the rest of the study. The finding that SDMA was not different at one month while urea, inosine and the fatty acids mentioned above were all changed suggests that SDMA was controlled through a different function, reflecting an adaption to the dietary intake rather than a mass action response.

This research was supported by Hill's Pet Nutrition and IDEXX Laboratories.
An avian model of nutrition and embryonic development: the requirement for cholesterol
Authors: Hsu Kun-Nan , Austic R. E.
Published: June 2021

The object of this study was to determine the need for cholesterol (C) in developing chicken embryos. Yolk was removed from eggs at 3 d of incubation (IB), and replaced by experimental yolk at 5 d. Cholesterol‐reduced egg yolk (CREY) lacking yolk granules was produced by beta‐cyclodextrin precipitation of C. Experiments (Exp) were done to (i) assess the importance of yolk granules for survival of embryos, (ii) to determine the effect of C on growth, survival, tissue C concentrations, and post‐hatch development. In Exp 1, embryo survival was markedly improved when yolk granules were added to CREY (i.e. CREYg). In Exp 2 and 3, embryos received C in levels of 2.3, 4.6, 12.1, and 13.5 mg of C/g (Groups (G) 1, 2, 3, and 4, respectively). G 4 consisted of intact eggs. The survival of embryos to 20 d IB was lowest (P<0.05) for G 1 in both Exp. Body wt. were low in G 1 and 2 (P<0.05). Liver C in G 1 and G 2 were reduced 96 and 85% in Exp 2, and 88 and 80% in Exp 3 (P<0.05). In Exp 4, the results for brain and liver C were consistent with those in Exp 2 and 3. Heart C was reduced in G 1 and 2 (P<0.05). In Exp 5, yolk C ranged from 6.8 to 19.8 mg/g: chickens that hatched matured at normal ages and were able to reproduce. In conclusion, the results suggest that more than 4.6 mg of C/g yolk is needed for normal growth. This avian model may be of value in the study of embryo nutrition, independent of maternal influence, on postnatal growth, physiological development, and health.
Supply and Demand Constraints in the Delivery of a National Food by Prescription Program for HIV+ Individuals in Ethiopia
Authors: Coates Jennifer , Stobaugh Heather , Bontrager Elizabeth
Published: June 2021

Few studies have examined the effect on health and nutrition outcomes of food offered “by prescription” to people living with HIV/AIDS. Likewise, the implementation processes conducive to achieving significant impacts and constraints to program effectiveness are not well documented. This study examines the delivery and utilization of a therapeutic Plumpy'nut ration as part of the Ethiopia National Food by Prescription (FBP) Program. Quantitative monitoring data from FBP clinic sites were analyzed to examine the proportion of patients attending ART and FBP appointments and receiving drugs and nutrition rations. 130 male and female FBP participants and medical staff were engaged in focus group discussions using participatory methods at 8 FBP program health facilities to examine factors related to program participation, adherence, and perceived changes in health and nutrition status. Approximately 50% of rations were reportedly consumed by the HIV+ index patient. Pressure to sell and share rations, negative ration side‐effects, transportation difficulties, stigma, and additional burden on health workers emerged as constraints to efficient implementation. Attention to often under‐utilized monitoring data can enable the timely resolution of critical implementation bottlenecks, leading to greater program effectiveness.
Sustainability of Backyard Gardening to Improve Nutrition in HIV‐affected Individuals in Swaziland
Authors: Malinga Steven , Magagula Celinhlanhla , Sibiya Thulani , Benson Shelby , Ramlal Roshan T
Published: June 2021

Objective

The CDC‐funded Community Linkages Project aims to improve nutrition of HIV‐affected individuals through backyard gardening in Swaziland.

Methods

Quantitative and qualitative data collected from February 2010 to September 2011 were used to assess the sustainability of backyard gardening to improve nutrition among those affected by HIV. Individuals included those at high risk of malnutrition due to comorbidities such as HIV and tuberculosis. The project provided training, garden tools, seeds of six vegetables and ongoing supervision to 355 individuals establishing backyard gardens. Individuals from dry and humid climates, across 14 clinics, who did not own a garden and were willing to participate, were selected and monitored. Individuals were categorized according to garden progress as follows, stage 1: land clearing and preparation, stage 2: seedbed preparation and transplanting, stage 3: weeding, watering, pest and disease control, and stage 4: harvesting.

Results

191 individuals were evaluated; others being lost to death or relocation. Of 191, 85% sustained their garden after a year (stage 3 or 4). Interviews showed benefits included many teaching neighbors the practice and expanding gardens to larger areas.

Conclusion

In a country where food and micronutrient supplements are becoming less popular due to issues of sustainability and cost, backyard gardens are a viable alternative.
Phosphorylation of cytokeratin fibers alters their response to mechanical extension
Authors: Fois Giorgio , Weimer Michael , Felder Erika T , Zheng Xing , Tobias Busch , Seufferlein Thomas , Dietl Paul , Felder Edward
Published: June 2021

Cytokeratin (CK) fibers protect epithelia from mechanical stress. High loads of mechanical strain lead to overextension of the CK‐fibers, discernible by wavy CK‐fibers in living cells after the strain is released. As mechanical properties of CK‐fibers are modulated by phosphorylation, we investigated the effect of phosphorylation on the waviness of overextended CK‐fibers.

Using FP‐labeled CK8/18 constructs and a cell stretching device, we found that wavy CK‐fibers were prevented by treatment with Staurosporin (unspecific kinase inhibitor), by depleting the cells of ATP and with CK‐mutants that abrogate phosphorylation at specific sites. Interfering with actin/myosin had a similar effect. The time that it took the CK‐fibers to regain their taut structure was not affected by phosphorylation but clearly depended on intact microtubuli.

Further experiments will identify the signaling pathway for CK‐fiber dephosphorylation and will demonstrate if the mechanical stimulation itself triggers this pathway ‐ possibly as an adaptive mechanism to protect the CK‐fibers from mechanical damage.

This work was supported by the DFG, grant DI 1402/1‐1 and DI1402/3‐1, and the Baden‐Württemberg Stiftung, grant P‐LS‐Biomat/03.
Cardiovascular complications among African Americans with HIV
Authors: Meshack Angela , Oyekan Adebayo , Agu Ngozi
Published: June 2021

Focus groups were conducted among 50 African American adults with HIV to investigate their perceived risks for cardiovascular disease and perceived benefits and barriers to living a heart healthy lifestyle. In addition, anthropometric measures were obtained (height, weight, blood pressure) and participants completed a written cardiovascular disease risk profile. Analysis of the cardiovascular risk profile data revealed that 48 percent of participants were current smokers 58 percent reported they had hypertension; 50 percent reported hypercholesterolemia; 30 percent that their mother or sister was diagnosed with a heart attack before age 65; and 16 percent as having diabetes. The computed body mass index of 80 percent of participants was categorized as overweight to obese. Participants were aware of cardiovascular risk factors, shared beliefs about the negative effects of HIV on cardiovascular health, and expressed barriers and facilitators to cardiovascular disease risk reduction. The findings suggest that some information has been disseminated about cardiovascular disease to HIV patients, but there is still a lack of information concerning the possible link between HIV disease, HIV medications, and cardiovascular disease. More cardiovascular disease risk reduction programs are needed for persons living with HIV. This research is supported by the National Heart, Lung, and Blood Institute.
Antagonism of CB1‐mediated discriminative‐stimulus effects by CB1 neutral and inverse‐agonist antagonists
Authors: Kangas Brian D. , Vemuri Kiran , Thakur Ganesh , Makriyannis Alex , Bergman Jack
Published: June 2021

CB1 inverse‐agonist antagonists (e.g., rimonabant) have behavioral effects that limit their clinical utility. Recent data suggest that CB1 neutral antagonists may not have rimonabant‐like effects, raising the possibility of improved clinical utility. The present studies were conducted to compare how inverse‐agonist antagonists (SR141716A, AM 6538) and neutral antagonists (AM 4113, AM 6545, AM 6527) modify the discriminative‐stimulus effects of the CB1 full agonist AM 4054 (0.01 mg/kg, i.m.) in nonhuman primates (n=4). Results show that each antagonist except AM 6545 produced dose‐related rightward shifts in the AM 4054 dose‐effect curve. Also, antagonist effects of AM 6538 were evident up to 4 days following administration. Pretreatment with AM 6545 (10 mg/kg acutely or daily for 5 days) did not greatly alter AM 4054 discrimination. These results show that: 1) both CB1 neutral and inverse‐agonist antagonists, while differing in potency, can surmountably antagonize CB1 discrimination and 2) AM 6538 has long‐lasting CB1 antagonist actions. The ineffectiveness of AM 6545 agrees with previous reports of its limited brain penetration. The present data indicate that CB1 neutral antagonism is an effective strategy for blocking addiction‐related effects of CB1 agonists, and support the idea that it may be useful in managing cannabis‐related disorders. (Supported by DA023142 and DA007252)
Newly Designed and Improved Human Soluble Epoxide Hydrolase Inhibitors for Neuropathic pain
Authors: Lee Kin Sing Stephen , Liu Jun-Yan , Inceoglu Bora , Wagner Karen , Dong Hua , Rose Tristan E. , Yang Jun , Morisseau Christophe , Hammock Bruce D.
Published: June 2021

Soluble epoxide hydrolase (sEH, EC 3.3.2.10) is responsible for metabolism of endogeneously derived fatty acid oxiranes, epoxyeicosatrienoic acids (EETs) to the corresponding diols, dihydroxyepoxyeicosatrienoic acids (DHETs). EETs are the products of arachidonic acid by cytochromes P450 epoxygenases and are key modulators involved in inflammation, pain and blood pressure. Inhibition of sEH that results in an increase in endogenous EETs, which have a significant effect on resolving inflammation and high blood pressure in rodent models. Therefore, the sEH has emerged as a pharmaceutical target for hypertension, inflammation and organ‐protection. Recently, we have demonstrated that elevation of epoxy‐fatty acid through sEH inhibition is able to resolve diabetic neuropathic pain in rodent.

Over last decade, a series of N,N′‐disubstituted ureas has been developed to study the physiological effects of sEH inhibition in various murine and rodent models and their beneficial effects in several animals disease models has been demonstrated. To better study the effect of epoxy‐fatty acids on health, a set of new compounds with dramatically potency, a better solubility and an improved pharmacokinetic profiles has been synthesized, their PK‐ADME and efficacy on diabetic neuropathic pain will be determined and the results will be presented and discussed here.
Measuring chemotherapy‐induced neuropathic pain and attenuation by cannabidiol using pain‐stimulated and paindepressed models in female C57Bl/6 mice
Authors: Pavlenko Khristina , Neelakantan Harshini , Walker Ellen A. , Ward Sara Jane
Published: June 2021

Paclitaxel (PAC) is associated with a chemotherapy‐induced peripheral neuropathy that can lead to the cessation of treatment in cancer patients, even in the absence of alternate therapies. Rodent models of PAC‐induced neuropathic pain have focused on stimulus‐evoked sensitivity measurements following systemic dosing of PAC, while the effect of chemotherapy‐induced neuropathy on pain‐depressed behaviors has not been characterized. Also, the phytocannabinoid cannabidiol (CBD) can reverse other types of neuropathic pain, but its effect on PAC‐induced neuropathy has not previously been investigated. We investigated the effect of CBD on PAC‐induced mechanical allodynia and PAC‐induced suppression of palatable food consumption. PAC administration (4.0 – 8.0 mg/kg × 4 injections, IP) led to the onset of mechanical allodynia as well as a suppression of palatable food consumption. Treatment with CBD (5.0 mg/kg IP) prevented the development of PAC‐induced mechanical allodynia and reversed PAC‐induced suppression of food consumption. The ability of CBD treatment to block PAC effects was attenuated by pretreatment with the 5‐HT1A antagonist WAY100635 (1.0 mg/kg IP), but not the CB1 antagonist SR141716 (3.0 mg/kg IP). In conclusion, CBD blocks PAC‐induced neuropathic pain using pain‐stimulated and pain‐depressed models, and these effects may be mediated by stimulation of 5‐ HT1A receptors.
Inhibition of neuropathic pain via dynorphin analog LYS 1044
Authors: Faridian Martin Kosar , Sandweiss Alex Jordan , Podolsky Alex , Lee Yeun-Sun , Vanderah Todd
Published: June 2021

Neuropathic pain commonly results from damaged nerves or tissue incorrectly sending continuous excitatory signals to nociceptors, resulting in chronic pain. Elevated levels of dynorphin A and bradykinin have been linked with chronic neuropathic pain. Furthermore, selective bradykinin receptor antagonists have shown to reduce hyperalgesia; blocking actions of dynorphin while continuing to allow low levels of bradykinin activity. LYS 1040 is a novel dynorphin analog that we have demonstrated efficacy of in the spinal nerve ligation (SNL) model in rats. Rats underwent (SNL) of the lumbar 5 and 6 nerves to stimulate neuropathic pain. Post injury rats were baselined to measure their state of tactile alloydynia (2.68g SEM+/−1.25) and thermal hyperalgesia (18.9s SEM+/− 0.54). After intrathecal injection of the drug, Von Frey testing (measuring mechanical allodynia) showed antiallodynia at all time points peaking at 100 minutes; 9.39g (SEM +/− 1.92) threshold, while the vehicle at 100 minutes withstood 3.71g (SEM +/− 0.60). IR testing showed antihyperalgesia at all points peaking at 40 minutes; 28.43s (SEM +/− 2.24), compared with vehicle treated animals which withstood 14.64s(SEM+/−1.82) at 40 minutes. Rotarod testing showed no inhibition of motor control for 120 minutes; LYS1044 ( 2.77m SEM+/− .18) as compared to vehicle (2.48m SEM+/− .58).
Adaptor Protein Complex‐2 and epsin‐1 mediate Protease‐activated Receptor‐1 internalization via phosphorylation‐ and ubiquitination‐dependent sorting signals
Authors: Chen Buxin , Dores Michael R. , Grimsey Neil , Canto Isabel , Barker Breann L. , Trejo JoAnn
Published: June 2021

Signaling by protease‐activated receptor‐1 (PAR1), a G protein‐coupled receptor (GPCR) for thrombin, is tightly regulated. PAR1 desensitization is mediated by β‐arrestins. However, internalization of PAR1 occurs through a clathrin‐ and dynamin‐dependent pathway independent of β‐arrestins. PAR1 displays two modes of internalization. Constitutive internalization of unactivated PAR1 is mediated by the clathrin adaptor protein complex‐2 (AP‐2). However, AP‐2 depletion only partially inhibits agonist‐induced internalization of PAR1, suggesting a function for other clathrin adaptors. We now report that AP‐2 and epsin‐1 are both critical mediators of agonist‐stimulated PAR1 internalization. We show that ubiquitination of PAR1 and the ubiquitin‐interacting motifs of epsin‐1 are required for epsin‐1‐dependent internalization of activated PAR1. AP‐2 regulates activated PAR1 internalization via recognition of distal C‐tail phosphorylation sites rather than the canonical tyrosine‐based motif. We further demonstrate a new function for AP‐2 in modulating the signaling responses of activated PAR1. Thus, AP‐2 and epsin‐1 are both required to promote efficient internalization of activated PAR1 and recognize discrete receptor sorting signals. This work was supported by National Institutes of Health Grant and a University of California Tobacco‐related Disease Research Program Exploratory Award.
Proteolytic activation of PAR1‐PAR2 heterodimer by thrombin promotes beta‐arrestin dependent ERK1,2 signaling
Authors: Lin Huilan , Trejo JoAnn
Published: June 2021

Thrombin cleaves and activates PAR1, but not PAR2. However, cleavage of the PAR1 N‐terminus by thrombin exposes a tethered ligand domain that can transactivate PAR2. Here, we show that PAR1 and PAR2 form a stable heterodimer, which co‐internalizes after thrombin stimulation. Thrombin activation of PAR1 also induces β‐arrestin recruitment and co‐internalization that requires co‐expression of PAR2. Cleavage of a signaling‐deficient PAR1 L258P mutant by thrombin is also sufficient for PAR2‐dependent β‐arrestin recruitment, suggesting that the tethered ligand domain of PAR1 activates PAR2. However, direct activation of PKC induces PAR2 internalization but not β‐arrestin recruitment. β‐arrestins can function as scaffolds and promote activation of ERK1,2 signaling. To assess the function of PAR1‐PAR2 recruitment of β‐arrestins we examined its capacity to promote ERK1,2 signaling. In cells expressing the PAR1‐PAR2 heterodimer, thrombin induced ERK1,2 signaling was markedly enhanced and prolonged compared to control cells not expressing β‐arrestins or cells expressing PAR1 alone. Thus, thrombinstimulated cellular signaling is modulated by PAR1 dimerization with PAR2. The mechanisms responsible for PAR1‐PAR2 dimer formation are under investigation. Supported by NIH R01 GM090689.
G Protein Coupled‐Receptor Kinase 5 is Localized to Centrosomes and Regulates Cell Cycle Progression
Authors: So Christopher H. , Michal Allison M. , Beeharry Neil , Shankar Haripriya , Mashayekhi Rouzbeh , Yen Timothy J. , Benovic Jeffrey L.
Published: June 2021

G protein coupled‐receptor kinases (GRKs) are important regulators of G protein coupled‐receptor function and mediate receptor desensitization, internalization and signaling. While GRKs also interact with and/or phosphorylate many other proteins and modify their function, relatively little is known about the cellular localization of endogenous GRKs. We find that GRK5 co‐localizes with gamma‐tubulin, centrin and pericentrin in centrosomes. The centrosomal localization of GRK5 is observed predominantly at interphase and although its localization is not dependent on microtubules, it can mediate microtubule nucleation of centrosomes. Knockdown of GRK5 expression leads to G2/M arrest, characterized by a prolonged G2 phase, which can be rescued by expression of wild type but not catalytically inactive GRK5. This G2/M arrest appears to be due to increased expression of p53, reduced activity of aurora A kinase and a subsequent delay in the activation of polo‐like kinase 1. Overall, these studies demonstrate that GRK5 is localized in the centrosome and regulates normal cell cycle progression. This work was supported by the National Institutes of Health.
Designing Spatiotemporal Regulators for PKA in Prostate Cancer
Authors: Kennedy Eileen J. , Lewandowski Megan , Hanold Laura , Ton Norman , Patel Fenil , Patel Viral
Published: June 2021

A Kinase Interating Protein 1 (AKIP1) is highly upregulated in prostate cancer and can mislocalize Protein Kinase A (PKA) by translocating it from the cytoplasm to the nucleus. Further, AKIP1 acts as a scaffold to stabilize interactions between PKA and other proteins, thereby influencing PKA‐mediated signaling. PKA also plays an important role in late‐stage, androgen‐independent prostate cancer by promoting androgen‐dependent transcription and nuclear localization of the androgen receptor. While it is apparent that AKIP1 plays an important regulatory role for PKA, the specific changes that are mediated by AKIP1 in prostate cancer remain largely unknown. Therefore, it is of great importance to understand the biological implications of nuclear PKA activity and how this relates to androgen receptor localization and signaling in late‐stage prostate cancer. To address this, we have developed novel peptide‐based chemical biology tools that act to disrupt protein‐protein interactions as a means to interrogate AKIP1 activity in the context of prostate cancer cells. By applying these chemically stabilized compounds, we can elegantly and selectively disrupt interactions between PKA and AKIP1 while leaving the catalytic activity of PKA intact, thereby enabling us to interrogate the role of AKIP1 in prostate cancer as well as uncover its regulatory role on androgen receptor signaling.