bluuebunny/crossref_metadata_2025 huggingface dataset:
Nitrogen balance adapts to changes in habitual protein intake within a few days in Japanese male athletes
Authors: Yokota Yukari , Tatsuta Wakako , Kamei Akiko , Kondo Emi , Kawahara Takashi , Okamura Koji
Published: June 2021

Eighteen healthy adult male Japanese athletes consumed meals containing 1.5 g/kg BW/day (P1.5) and 2.5 g/kg BW/day (P2.5) protein for 10 days, in a different order during a 20 day study to investigate the effect of a sudden change of habitual protein intake on nitrogen balance. Eleven subjects were consumed P1.5 from day 1 to 10 then switched to P2.5 from day 11 to 20 (P1.5/2.5), while 7 subjects were tested in the opposite order (P2.5/1.5). Urine samples were collected during the first and the last 3 consecutive days both before and after the meal switch; days 1–3 and days 8–10 of the 1st meal, days 11–13 and days 18–20 after the meal switch. Urinary nitrogen excretion significantly changed immediately in both groups after changing meals and did not change thereafter. The average of each urine sample period was 13.5, 13.4, 17.9 and 19.9 g/day for P1.5/2.5, and 17.5, 18.4, 14.1 and 13.5 g/day for P2.5/1.5. The nitrogen balance showed similar change as nitrogen excretion; 10.9, 11.2, 17.7 and 16.1 g/day for P1.5/2.5, and 16.1, 16.2, 9.2 and 11.1 g/day for P2.5/1.5. Therefore, both urinary nitrogen excretion and the nitrogen balance seems to adapt to both an increase and a decrease in the habitual protein intake within a few days in Japanese male athletes.
Use of omega‐3 fatty acid rich algae and their oil as a feed supplement for dairy cattle
Authors: Stamey Jennifer , Shepherd Dare , de Veth Michael , Corl Benjamin
Published: June 2021

Our objective was to examine use of algal biomass as an omega‐3 fatty acid‐enriched ration supplement for dairy cattle. Rations of four lactating cows were supplemented with 1X or 0.5X rumen protected (RP) algal biomass supplement, 1X RP algal oil supplement, or no supplement in a 4×4 Latin Square design. Supplements were lipid encapsulated (Balchem Corp., New Hampton, NY). 1X supplements provided 29 g/d docosahexaenoic acid (DHA). Supplementing dairy rations with rumen protected algal supplements did not affect feed intake, milk yield, or milk composition (P>0.05). Both 0.5X and 1X RP algae supplements increased (P<0.01) daily milk fat yield of DHA (0.5 and 0.6±0.10 g/d, respectively) compared to 1X RP oil (0.3±0.10 g/d), but all supplements were greater (P<0.01) than control (0.1±0.10 g/d). Yield of trans‐18:1 fatty acids in milk fat was also increased by supplementation, suggesting supplements may have influenced rumen microflora. Trans‐11 18:1 yield (13, 20, 27, and 15±3.0 g/d for control, 0.5X RP algae, 1X RP algae, and 1X RP oil, respectively) was greater for supplements than control (P=0.05). Rumen protected algal biomass provided better DHA yield than algal oil.
Dietary diversity scores predict dietary micronutrient adequacy among school children in rural Kenya
Authors: Gewa Constance Awuor , Murphy Suzanne P. , Neumann Charlotte G.
Published: June 2021

We explored the ability of dietary diversity scores to predict micronutrient intake among school children in rural Kenya. Three non‐consecutive 24‐hour recalls were administered to assess 520 school children's dietary intake in May‐August 1998. Diversity scores were calculated as counts of the number of food groups reported. Food groups included cereals/tubers; dark‐green leafy vegetables; vitamin A‐rich fruits/vegetables; vitamin C‐rich fruits/vegetables; other fruits/vegetables; dairy; meat/fish/poultry and legumes/nuts. Three diversity scores were constructed based on minimum amount required to make a food group count: DDS for a minimum intake of 1g, DDS‐15 for a minimum intake of 15g, and DDS‐V with variable minimum intake levels across food groups. The probability approach was used to estimate dietary adequacy of 12 vitamins and minerals. The mean probability of adequacy, ranging from 0 to 1 and a mean of 0.59 ± 0.07, was used as a measure of overall dietary adequacy. All three diversity scores were significantly associated with MPA. Correlations ranged from 0.46 for DDS to 0.51 for DDS‐V. Thus, all three measures of diversity may be used as indicators of dietary nutrient adequacy, although DDS‐V performed somewhat better than DDS or DDS‐15. The analysis contributes to the continued search for simpler indicators of dietary micronutrient adequacy and is the first among school children in Africa.
Hydration status of collegiate female basketball players at Southern Utah University
Authors: Schmidt Matthew Christopher , Walsh Mykel , Frehner Christopher
Published: June 2021

The purpose was to determine the hydration status in eleven collegiate female basketball players and examine if education would improve hydration status. Urines were collected before and after practice. Urine specific gravities (USG) were used to determine hydration status. Four urine samples were collected in the preseason and averaged. Athletes were unaware of these measurements. It is recommended that athletes begin practice at an USG ≤1.020. The team's average preseason USG was 1.020 at the beginning and 1.024 at the end of practice. Indicating that athletes came to practice minimally dehydrated and left significantly dehydrated. Athletes were aware of their preseason USGs. Education and strategies on proper hydration were provided and athletes were notified of the next USG measurements taken during the regular season. Average USGs for this measurement were 1.017 and 1.018 indicating that athletes entered and left practice minimally dehydrated. A third measurement was taken later in the season and athletes were unaware. This was to test if the improved hydration status in the second measurement would continue. The team's final average USG was 1.018 at the beginning and 1.023 at the end of practice. These findings suggest that education and strategies improved hydration status throughout the season or that subjects better prepared for urine test when they were aware of the measurement.
Results from the Development Research‐Intense Assessment Toolkit for a Sensory Nutrition Education Project Introducing Head Start Children to Exotic Fruits
Authors: Kannan Srimathi , Smith Rebecca L. , Foley Christine E. , Severin Suzanne
Published: June 2021

The FruitZotic (FZ) pilot research project aims to examine the feasibility of the development and testing of research instruments for sensory based exotic fruit nutrition education on Western Massachusetts Head Start children's willingness to try exotic fruits. Head Start preschoolers, ages 3–5 (n=245 children) were introduced to FZ on the basis of Head Start learning domains. During 2008–2010, a six‐week lesson series was implemented in two‐six classrooms per season (fall, and spring) over a 6‐week period. Pedagogies included book reading, sensory exploration and fruit tasting. New fruits (coconut, pineapple, kiwi) were selected under the theme “Fruits with Hair.” Book readings, craft activities, “circletime” tasting and pre‐ and post fruit cup waste measures were incorporated. FZ expanded with “Fruits from Afar” (mango, pomegranate, papaya, starfruit) and “Fruit Expo”. A modified rating scale [0–5, not willing to taste, willing to lick etc.] was designed to tracked changes in tasting progress. Baseline willingness assessment was introduced for evaluating classroom‐level changes in willingness to try exotic fruits. The 2‐year pilot project contributed to the design of research‐robust instruments to assess willingness to taste exotic fruits in children's natural learning environment. Project was funded by UMASS Commonwealth College Honors Grant and the Department of Nutrition UMASS Faculty Start‐up Funds.
Nutrition education and counseling during pregnancy: a systematic review
Authors: Girard Amy Webb , Olude Oluwafunke Abigail , Imhoff‐Kunsch Beth , Ramakrishnan Usha , Martorell Reynaldo
Published: June 2021

Nutrition education and counseling (NEC) is a common strategy used to improve maternal nutrition during pregnancy. Using modified GRADE (Grading of Recommendations Assessment, Development and Evaluation) and CHERG (Child Health Epidemiology Reference Group) approaches, we systematically reviewed the scientific evidence of the impact of NEC delivered during pregnancy on maternal diet, nutritional status, and pregnancy outcomes. Of 3877 titles identified through databases and hand searching, 52 were abstracted. Of these, 25 met final inclusion criteria and were included in this review. Three categories of NEC delivery emerged, namely 1) only NEC (n=14); 2) NEC with enhanced education or psychosocial support (n=3); and 3) NEC with nutrition support (n=9). In developed countries (n=17 studies), consistent and positive impacts of NEC on dietary intakes (n=12) were observed regardless of delivery category. In developing countries (n=9), NEC either alone or with nutrition support improved anemia outcomes (n=5). However for weight gain (n=2) and birth weight (n=2) consistent and positive impacts were only observed when NEC was provided with nutrition support. Substantial variation in exposure and outcome variables precluded meta‐analysis. Funded by the Bill and Melinda Gates Foundation
The hepatic Igf2/H19 locus is not altered in 1‐day old pups born to obese‐prone Sprague‐Dawley rats fed a low protein diet containing adequate folic acid
Authors: Uthus Eric O , Claycombe Kate J , Johnson William T
Published: June 2021

Gong et al. (Epigenetics, 2010) found, using diets low in folic acid, that compared to an 18% protein diet a 9% protein diet fed to pregnant Sprague‐Dawley rats resulted in increased Igf2 and H19 gene expression in the liver of day 0 male offspring. In addition DNA methylation in the Imprinting Control Region (ICR) of the Igf2/H19 locus was significantly increased. The ICR contributes to controlling gene expression of the Igf2/H19 locus in a methylation‐dependent manner. We fed obese‐prone Sprague‐Dawley females an AIN‐93 based diet containing adequate folic acid and either 8% or 20% protein diet for 3 wk. They were then bred with chow‐fed obese‐prone males and fed their respective diets through pregnancy. At birth, litters were culled to 4 male and 4 female; each litter (by sex) was considered as N=1. At 1 day, weights (means±SD) were: 8% males, 5.15±0.42 (N=12); 20% males, 5.32±0.41 (N=13); 8% females, 4.74±0.62 (N=13); 20% females, 4.93±0.25 (N=13). Hepatic DNA methylation status of the ICR, which includes 4 CTCF binding sites, was determined by pyrosequencing. Real‐time PCR was used to determine liver mRNA expression (relative to Actin) of Igf2, H19 and methyl CpG‐binding domain 2 (Mbd2). Regardless of sex of the pups we found no effect of diet on DNA methylation status of the ICR or expression of Igf2, H19 or Mbp2. Thus, these results suggest that low protein by itself does not contribute to an alteration in the Igf2/H19 locus.
Prenatal Exposure to Ethanol and the Epigenetic Regulation of Serotonin Transporter (Slc6a4) and Glucocorticoid Receptor (Nr3c1) Genes
Authors: Ngai Ying Fai Tiffany , Sulistyoningrum Dian C , Innis Sheila M , Weinberg Joanne , Devlin Angela M
Published: June 2021

Fetal Alcohol Spectrum Disorder (FASD) causes a range of cognitive impairment however the molecular aetiology underlying FASD is not fully understood. We hypothesize that developmental programming of gene expression via epigenetic mechanisms may contribute to FASD. We studied the expression of the glucocorticoid receptor (Nr3c1) and the serotonin transporter (Slc6a4) in the hippocampus and hypothalamus of young adult rats exposed prenatally to ethanol. Female Sprague‐Dawley rats were fed a liquid ethanol diet (E), a pair‐fed liquid control diet (PF), or lab chow (C) throughout pregnancy and the adult offspring were studied at 55 days of age. Female rats from E dams had lower (P<0.05) Nr3c1 mRNA levels in hippocampus and lower (P<0.01) Slc6a4 mRNA levels in hypothalamus than female rats from PF dams. Male rats from E dams had higher (P<0.05) Slc6a4 mRNA levels in hippocampus and lower (P<0.01) Slc6a4 mRNA levels in hypothalamus than male rats from C dams. Further analysis of the methylation status of 23 CpG sites in the 5′ regulatory region of Slc6a4 in hypothalamus showed lower (P<0.05) methylation in female and male rats from E dams compared to female and male rats, respectively, from C and PF dams. These findings suggest that prenatal exposure to alcohol may affect the developmental programming of gene expression in hypothalamus via epigenetic mechanisms.
A maternal low protein diet programs glucose and fatty acid metabolism differentially in adult male and female mouse offspring
Authors: Plosch Torsten , van Straten Esther M.E. , van Dijk Theo H. , Bloks Vincent W. , Verkade Henkjan J. , Kuipers Folkert
Published: June 2021

Nutritional conditions during fetal life can influence the risk to develop the metabolic syndrome in adult life (‘metabolic programming’). We aimed to establish a mouse model of metabolic programming focusing on the effects of a maternal low protein diet during gestation on glucose and lipid metabolism in the adult offspring. Pregnant C57BL/6J mice received a control or a low protein diet throughout gestation. Offspring received a low fat diet or a high fat diet from 6–22 weeks of age. Glucose metabolism was studied with a whole‐body‐glucose test using [6,6‐2H]‐glucose. Hepatic gene expression was characterized by microarray. Maternal low‐protein‐diet did not affect glucose metabolism in male offspring. Female offspring from normal‐protein fed dams was relatively resistant to diet‐induced metabolic dysregulation. Maternal low‐protein‐diet during gestation led to deteriorated insulin sensitivity upon high‐fat feeding in female offspring. Microarray analyses showed a sex‐specific effect of the maternal diet. We conclude that, in mice, maternal protein restriction during gestation does not change the glucose response to a high fat diet in male offspring. However, it changes fatty acid and glucose metabolism in female offspring in such a way that it resembles male metabolism. Supported by the Netherlands Heart Foundation, grant 2004T048.
Maternal exercise during pregnancy in swine programs gene expression in intestine of offspring
Authors: Arentson Emily , Buhman Kimberly K , Cabot Ryan , Newcomer Sean , Donkin Shawn S
Published: June 2021

Metabolic programming events induced by changes in maternal environment during gestation have marked effects on fetal outcomes. Aerobic exercise is known to alter maternal metabolism, however the consequences on fetal metabolic programming are unknown. To determine the effects of aerobic exercise training during pregnancy on metabolic programming of the offspring, pregnant commercial sows were limit fed a chow diet and exercised on a treadmill (EXER; n=5) for 30–45 minutes per day, 5 days a week at 65 –85% of maximal heart rate or allowed normal activity level (SED; n=4) for 15 weeks. Total RNA isolated from duodenum (DUO), jejunum (JEJ) and ileum (IL) mucosal of two piglets per litter was analyzed for abundance of expression for key transcripts involved in energy metabolism. PPAR gamma coactivator‐1α mRNA abundance was elevated (P < 0.01) in the JEJ and IL of EXER vs. SED offspring (JEJ: 2.24 vs. 1.01, IL: 2.03 vs. 0.81, CI [0.04, 0.43]) and carnitine palmitate transporter 1α was increased (P < 0.01) in the EX vs. SED offspring (1.34 vs. 0.93, CI [0.99,1.23]). The data indicate programming of genes for intestinal energy metabolism in offspring in response to maternal aerobic exercise during pregnancy.

Grant Funding Source: NIDDK
A Comparative Study of Eating Habit and Food Intake of Korean Women with Gestational Diabetes According to Early Postpartum Glucose Level
Authors: Park Sunmin , Park You Jeong , Kim Sung Hoon
Published: June 2021

Women with gestational diabetes mellitus(GDM) are at high risk for Type 2 diabetes mellitus(DM) and cardiovascular disease(CVD). However, little is known about dietary patterns of GDM women after delivery. We compared the eating habits and food intakes of Korean GDM women in the early postpartum period according to postpartum glucose tolerance. After performed a 75g oral glucose tolerance test in 184 women with GDM at 6–12 weeks after delivery, they were divided into three groups; normal glucose tolerance(NGT, n=100), prediabetes(n=73), and DM(n=11) according to ADA criteria. Eating habits and usual food intakes after delivery were investigated by survey questionnaires and 24‐h recalls with a trained dietitian. Biochemical parameters were measured at pre‐ and post‐delivery. Serum triglyceride concentration and blood pressure were higher in the DM group than the NGT group. Eating habits were not significantly different among the three groups. Animal fat consumption was higher in the prediabetes and DM groups than in the NGT group, and dietary cholesterol in the prediabetes group was higher than in the NGT group. The intake ratio of fat calories to total calories was also significantly higher in the prediabetes group. In conclusion, it may be important to educate women with GDM about the risks of excessive animal fat and cholesterol intakes during pregnancy and the postpartum period in order to prevent Type 2 DM and CVD.
Calcium intake in the United States from dietary and supplemental sources across adult age groups: new estimates from NHANES 2003–2006
Authors: Mangano Kelsey McKee , Walsh Stephen J , Insogna Karl L , Kenny Anne M , Kerstetter Jane E
Published: June 2021

Adequate lifelong calcium intake is essential in optimizing bone health. Recent National Health and Nutrition Examination Survey (NHANES) data were used to quantify variation in calcium intake across adult age groups, to relate these changes with energy intake, to assess differences in dietary calcium intake between supplemental calcium users and non‐users, and to evaluate associations between age and calcium density in the diet. This cross‐sectional analysis determined calcium and energy intake for 9,475 NHANES respondents during the 2003–06 period. Nutrient density was represented using calcium:energy ratios. When compared to the 19–30 y age group, median dietary calcium intake was lower in the 81+ age group by 23% in men (p<0.001) and by 14% in women (p=0.003). These reductions coincided with 35% and 28% decreases, respectively, in median energy intake (p<0.001). Although the frequency of calcium supplement use increased with age (p<0.001), among female supplement users, the decline in dietary calcium intake was greater than in non‐users (p=0.02). Calcium density in the diet increased relative to age in men and women (p<0.001); however, calcium:energy ratios were insufficient to meet target ratios inferred by adequate intake standards after age 50. Overall, although calcium supplementation and density were highest in older age groups, intakes remain insufficient in meeting recommended levels.

Grant Funding Source: Nutrition Pilot Grant Program of the Patrick and Catherine Weldon Donaghue Medical Research Foundation
Comparative Study of the Prevalence of Genetic Modification in Commonly Available Maize Products
Authors: Lanning Michael Clark , Thiery Megan , Cheeseman Kerry
Published: June 2021

Genetic modification of food crops for nutritional or other commercial reasons remains a polarizing topic. Arguments for and against the procedure are abundant in the literature. The purpose of this study has been to determine the overall prevalence of genetic modification in commercially available maize products, focusing specifically on popcorn products and comparing these results to previous work done in this lab on both whole kernel and processed corn products. Using standardized biochemical practices, DNA was extracted in duplicate from store bought popcorn samples and amplified through PCR to detect the CaMV 35S promoter and/or NOS terminator sequences (used in approximately 85% of genetically modified food products). Samples were run on 3% agarose gels, along with positive and negative GM controls, and visualized with ultraviolet light following ethidium bromide staining. Previous work with processed corn products (n=56) and whole kernel maize (n=16) resulted in evidence of genetic modification in approximately 75% and 56% of products, respectively. Initial results for popcorn samples seems to indicate a prevalence similar to that found in whole kernel maize, however further exploration is needed to reach a conclusive result. This work has been supported by a grant from Beta Beta Beta Biological Honor Society.
High fat diet‐induced obesity leads to proinflammatory response associated with higher expression of NOD2 protein
Authors: Kim Min Soo , Han Sung Nim
Published: June 2021

Obesity has been reported to be associated with low grade inflammatory status. We studied the inflammatory response of immune cells and signaling molecules associated with inflammatory responses in diet‐induced obesity. C57BL mice were fed diets containing 5% fat (Control) or 20% fat and 1% cholesterol (HFD) for 24 wks. Splenocytes (1 × 107 cells) were stimulated with 10 μg/mL of LPS for 6 or 24 hrs. Production of IL‐1β, IL‐6 and TNF‐α and expression levels of NOD2, STAT3, pSTAT3 were determined. Consumption of HFD resulted in significantly more weight gain (28.2±0.6g in HFD vs. 15.4±0.8g in control). After stimulation with LPS for 6 hrs, production of IL‐1β and TNF‐α were significantly higher in HFD group. After 24 hrs of LPS stimulation, splenocytes from HFD group produced significantly higher levels of IL‐6 (10.02 ± 0.66 ng/mL in HFD vs. 7.33 ± 0.56 ng/mL in control) and IL‐1β(121.34 ± 12.72 pg/mL in HFD vs. 49.78 ± 6.58 pg/mL in control). There were no significant differences in STAT3 and pSTAT3 expression levels between HFD and control groups. However, expression level of NOD2 proteins determined by western blot analysis was significantly higher in HFD group (60% higher in HFD group compared with control, p<0.05). NOD2 contributes to the induction of inflammation by activation of NF‐κB. These findings suggest that diet‐induced obesity is associated with increased inflammatory response of immune cells and higher expression of NOD2 may contribute to these changes.

Supported by Korea Research Foundation Grant funded by the Korean Government(KRF‐331‐2008‐1‐C00305) and the Basic Science Research Program funded by the Ministry of Education, Science and Technology (2009‐0063409).
Cocoa supplementation can reduce systemic inflammation and body weight gain in obese mice
Authors: Gu Yeyi , Lambert Joshua D.
Published: June 2021

In the present study, we investigated the effect of short‐term supplementation of cocoa powder (8% w/w, 10 weeks) on biomarkers of systemic inflammation and body weight gain in obese mice. Male C57BL/6J mice were initially randomized into two groups: low fat diet (LF: 10% kcal from fat, n= 12) and high fat diet (HF: 60% kcal from fat, n=46). At week 9, 23 obese mice from HF group were randomized into 8% cocoa‐ high fat diet (HFC: 59% kcal from fat). At the end of week 18, mice were euthanized and blood and tissues were collected. Our results indicate that cocoa supplementation for 10 weeks can reduce obesity induced systemic inflammation with decreases in the plasma proinflammatory cytokine interleukin ‐6 (IL‐6, 30.4%), monocyte chemoattractant protein 1 (MCP‐1, 25.2%), tumor necrosis factor‐alpha (TNF‐α, 17.4%), and an increase in the anti‐ inflammatory marker adiponectin (33.7%) compared with HF mice. Cocoa supplementation also significantly reduced body weight gain in high fat induced obese mice (HF=11.4g vs. HFC=9.5g) without affecting food intake. The results suggested that cocoa supplementation is beneficial in prevention of systemic inflammation and slowing body weight gain in obese mice. Other biochemical analyses are under investigation. Unsweetened cocoa powders were provided by Blommer Chocolate Co. (Chicago, IL). This study was supported by NIH Grant AT004678 (to JDL).
NOD1 activation induces proinflammatory gene expression and insulin resistance in 3T3‐L1 adipocytes
Authors: Zhao Ling , Hu Pan , Purohit Jaanki , Hwang Daniel H
Published: June 2021

Chronic inflammation is associated with obesity and insulin resistance. However, the underlying mechanisms are not fully understood. Pattern recognition receptors (PRRs) play critical roles in innate immune response. Here we report a role of Nucleotide‐binding Oligomerization Domain protein 1 (NOD1), a member of PRR, in adipocyte inflammation and insulin resistance. mRNA level of NOD1 is increased by 4 fold in differentiated 3T3‐L1 adipocytes compared with preadipocytes. Stimulation of NOD1 with a synthetic ligand Tri‐DAP induces proinflammatory chemokine MCP‐1, RANTES and cytokine TNF‐α and MIP‐2 mRNA expression and protein secretion. Moreover, stimulation of NOD1 with Tri‐DAP suppresses insulin signaling as revealed by attenuated Akt phosphorylation on Ser 473 and Thr 308 and subsequent insulin‐induced glucose uptake in 3T3‐L1 adipocytes. In addition, NOD1 mRNA is also increased upon differentiation of primary culture derived from human fat. Stimulation of NOD1 induces similar proinflammatory chemokine/cytokine mRNA expression and protein secretion. Together, our results suggest that NOD1 may be involved in adipose inflammation and insulin resistance. The work is supported by The University of Tennessee faculty start‐up fund and WHNRC USDA/ARS program funds.
Regulation of adipose tissue function and whole body metabolism by biglycan
Authors: Ward Meliza , Ajuwon Kolapo
Published: June 2021

Biglycan (Bgn) is a component of the extracellular matrix that can signal through toll‐like receptor 4, leading to increased inflammation. Therefore, increases in Bgn expression in adipose during obesity may contribute to obesity‐induced inflammation. In our study, we compared Bgn knockout mice (bgn‐KO) with wild‐type mice (WT) during high fat diet induced obesity. Mice of both genotypes were fed a low‐fat (LFD, 10% fat calories) or a high‐fat diet (HFD, 60% fat calories) for 10 weeks. Expression of Bgn was increased in the brown fat, mesenteric, and epididymal fat of WT mice on HFD when compared to WT mice on LFD (p<0.05). Wild type animals had a higher body weight than bgn‐KO (p<0.05). There was no difference in BMI between genotypes (p>0.05). Likewise, percent adiposity, expressed as weight of epididymal, subcutaneous and retroperitoneal fat pads over body weight was not different between genotypes (p>0.05). Wild type mice had higher levels of fasting insulin than bgn‐KO (p<0.05). Gene expression of IL‐6 was lower in the subcutaneous fat of bgn‐KO mice when compared to WT (p<0.05), while expression of adiponectin was higher in the epididymal fat of bgn‐KO mice (p<0.05). Our data indicate that bgn‐KO mice may have an improved metabolic and inflammatory phenotype, despite comparable adiposity relative to WT. This supports a role for Bgn in the regulation of insulin sensitivity and inflammation in adipose tissue.

Grant Funding Source: Purdue University
Seasonal variations in vitamin D status in Bangladesh: a preliminary look at the potential role of aerosol pollution
Authors: Roth Daniel E. , Al‐Mahmud Abdullah , Kimlin Michael , Arifeen Shams El , Raqib Rubhana , Black Robert E. , Baqui Abdullah H
Published: June 2021

Vitamin D deficiency is common in South Asia, but the causes are unclear. Particulate air pollution interferes with ultraviolet (UV) irradiation of the earth surface, but the effect of UV‐absorbing aerosols on vitamin D status is unknown. Associations among vitamin D status (serum 25‐hydroxyvitamin D, [25(OH)D]), UV radiation exposure and UV‐absorbing aerosols in Dhaka were explored based on observations of 34 non‐pregnant and 13 pregnant women in July‐Oct 2009, and 28 pregnant women in Feb 2010 (N=75). Average [25(OH)D] was lower in winter vs. summer/fall; among the pregnant women, mean [25(OH)D] was 12 nmol/L lower in Feb vs. Aug‐Oct (95% CI, 1 – 22 nmol/L). Seasonal differences in [25(OH)D] could be largely explained by variations in personal UV exposure estimated using polysulphone (PS) badges worn on the shoulder, averaged over 2–3 days within ~2 months of [25(OH)D] assessment (N=72). UV exposure by PS dosimetry and [25(OH)D] were both inversely associated with the estimated UV aerosol index (UVAI) for Dhaka, based on satellite‐generated data downloaded from an on‐line NASA database (OMI/Aura). The UVAI peaked in early spring (before the monsoon season), when average UV exposures by PS dosimetry were lowest. These preliminary observations suggest that UV‐absorbing aerosols may have important effects on UV‐induced cutaneous vitamin D synthesis in South Asia and merit further research.

Grant Funding Source: Center for Global Health, Johns Hopkins University; Department of International Health, Johns Hopkins Bloomberg School of Public Health
Chronic alcohol effects on NF‐κB genome‐wide binding dynamics during early onset of liver regeneration
Authors: PATRA BISWANATH , HOEK JAN B , VADIGEPALLI RAJANIKANTH
Published: June 2021

The objective of this study is to characterize effects of chronic alcohol consumption on NF‐κB mediated transcriptional regulatory network dynamics during the early phase liver regeneration after 70% partial hepatectomy (PHx) at 1 and 6h post 2/3rd PHx in the rat. Rats were fed liquid diet containing 36% of total calories derived from ethanol for 5 weeks, controls were pair‐fed isocaloric liquid diet. Genome‐wide NF‐κB binding targets were detected using Roche NimbleGen ChIP‐chip microarray platform using Chromatin Immunoprecipitated (ChIP) samples, several of which were validated by ChIP‐qPCR. Our results indicate that (1) chronic ethanol diet did not increase the genome‐wide binding activity of NF‐κB at the baseline state with ~3200 target gene promoters; (2) PHx induced significant increase in NF‐κB genome‐wide binding at 1h, more so in chronic ethanol samples (5300 genes) than in controls (4000 genes). At 6h post PHx, only the control samples showed a further increase in NF‐κB promoter binding ~5500 genes. Pathway analysis revealed that NF‐κB target genes specific to the chronic ethanol group participate in key processes such as DNA methylation, cell death, proteolysis, histone modification and regulation of cell cycle. We conclude that at least some of the chronic ethanol effects on liver regeneration are mediated by altered NF‐κB binding at specific pathway genes. Research Support: NIH AA018873, AA016919.
Exogenous oxytocin (OT) ameliorates pulmonary damage caused by Escherichia coli (E. coli) infection in chronically stressed rats
Authors: Stadnikov Alexander A , Kozlova Alina N , Klimushkin Aleksey V
Published: June 2021

This study examines the hypothesis that exogenous OT will reduce pulmonary damage caused by E. coli infection in stressed rats. Four groups of male albino rats were used as follows: 1) rats received an intratracheal (i.t.) injection of 0.2 mL E. coli suspension containing 1·109 bacteria/mL in saline; 2) rats were subjected to footshock (FS) stress for 1–11 days; 3) rats were subjected to FS stress for 7 days and then received an i.t. injection of E. coli; 4) rats subjected to FS stress had concurrently received i.m. injections of OT (0,02 IU/mL) for 7 days and then were injected with E. coli. The lungs, hypothalamus, and neurohypophysis were analyzed by light and electron microscopy, immunohistochemistry on days 1 to 11 (groups: 1–2) and days 8 to 11 (groups: 3–4). We found that an E. coli infection in chronically stressed rats caused a greater inflammatory response, cell death and fibrosis than were seen in non‐FS animals. The detrimental changes in the former coincided with increased inhibition of OT release from neurohypophysial terminals. The injections of OT reduced the number of apoptotic cells (~40%) and production of MMP‐9 (~75%), while it stimulated the expression of Bcl‐2, Clara cell protein (CC‐16), and lactoferrin in the lungs. Thus, we concluded that exogenous OT shows a therapeutic effect against E. coli‐induced lung injury under condition of chronic stress, when the release of endogenous neuropeptide is impaired.
Incidence of Complex Vascular Lesions (Plexiform‐Like Lesions) in the Lungs of 1 to 21‐Day‐Old Domestic Fowl
Authors: Wideman Robert F , Mason Grant , Hamal Krishna R , Cross David W , Rhoads Douglas D , Erf Gisela F , Anthony Nicholas B
Published: June 2021

Plexogenic arteriopathy rarely develops in existing animal models of pulmonary arterial hypertension (PAH). Rapidly growing chickens spontaneously develop PAH accompanied by the formation of plexiform‐like complex vascular lesions (CVL). The objective of this study was to evaluate the frequency with which at least one CVL per tissue section was observed in the lungs of chicks from lines selected for susceptibility (S) or resistance (R) to PAH. At 1, 7, 14 and 21 days of age (n ≥ 10 chicks per age and line) the CVL incidences were 33, 30, 35 and 50% for S chicks, and 20, 30, 28, and 25% for R chicks. The combined CVL incidences over days 1–21 did not differ significantly (P = 0.105) between S (36.4%) and R (25.5%) chicks. Early CVL formed when proliferating intimal cells occluded an arteriole lumen distal to branch points in muscular inter‐parabronchial arterioles that typically exhibited medial hypertrophy and intimal proliferation. Mature CVL in chicken lungs strikingly resemble published photomicrographs of plexiform lesions in the lungs of human PAH patients. The development of CVL in the lungs of young chicks suggests a complex interaction between pulmonary hemodynamics, vascular development and vascular pathology. Supported by NIH grant 1R15HL092517.
Development of a database for anatomical terms via web
Authors: Barros Mirna Duarte , Carrare Ana Paula G , Curcio Daniella Franco
Published: June 2021

INTRODUCTION

The international anatomical terminology is the foundation of the scientific terminology in the health field and its main purpose is to standardize the vocabulary employed in the identification of structures of the human body.

OBJECTIVE

To develop a database containing information of the anatomical terminology, translations, eponymous and clinical terms, with open access via web.

METHOD

The system was developed with interface to web environment and database in MySQL. The structure of the data respected the hierarchic order of the terms, allowing visualization of the term within its context. The system forecasts generation of reports with diverse configurations.

RESULTS

The first version of the system is concluded and in the stage of data feed. Beyond the data of the international anatomical terminology, the systems allows for the inclusion of eponymous, clinical terms and translations into other languages. The search may be performed by typing the term inside a search‐box or by hierarchic visualization. The access to the database via web provides agility to retrieve information as well as to implement updates that might be officially adopted.

CONCLUSIONS

The role of the anatomical terminology depends on its application in the didactic, clinical and scientific practice and an open database via web contributes to widen the access to its information and to spread its usage in the health field.
Identification and characterization of SARS‐CoV protein 3a and its interacting cellular proteins
Authors: Yu Chia‐Jung , Wu Hsu‐Ying , Chen Jian‐Hong , Hsu Yu‐Tien , Lee Fang‐Jen S.
Published: June 2021

The severe acute respiratory syndrome coronavirus (SARS‐CoV) protein 3a is a non‐structure glycoprotein of SARS‐CoV, which with multi‐functions in host cell, including induces apoptosis, leads to cell cycle arrest at G1 phase, forms ion channel, and causes Golgi fragmentation. However, the detail molecular mechanism response for functional modulations of protein 3a remains unknown. Herein, we found that expression of non‐tagged protein 3a in Vero E6 cells reduced the mitochondria membrane potential, caused Golgi fragmentation, and impaired the vesicle transport. Using in vitro pull‐down assay combined with mass spectrometry, we identified 23 potential protein 3a‐interacting cellular proteins. Based on biological classification, we selected 60 kDa heat shock protein (Hsp60) and SNAPs association protein (SAP) for further characterization. Biochemical fractionation indicated that Hsp60 protein level in mitochondria fraction was increased in protein 3a‐expressing cells. Overexpressed SAP was partially co‐localized with protein 3a and co‐immunoprecipitated with protein 3a. Collectively, our results implied that the effect of protein 3a on the function of mitochondria and the Golgi might through its interaction with Hsp60 and SAP, respectively.
Regulation of Cellular Transformation Genes by the Circadian Clock Protein Per1 in the Kidney
Authors: Gumz Michelle L. , Cheng Kit‐Yan , von Roemeling Christina , Copland John A. , Gilkes Janine
Published: June 2021

The importance of normal circadian rhythms is underscored by the known relationship between circadian disruption and increased cancer risk. The goal of the present study was to investigate the role of the circadian clock in the regulation of genes encoding products that function in cellular transformation. Microarray analysis was used to evaluate changes in gene expression following siRNA‐mediated knockdown of the circadian clock protein Per 1 in a murine renal collecting duct cell line (mpkCCDc14). Several hundred genes underwent significant changes in expression following Per1 knockdown, including 21 genes involved in cellular transformation. Cdkn1a and Cyclin E1, both cell cycle regulators, claudin 1 and 12, involved in tight junction formation, and sFRP1, a regulator of Wnt signaling and a tumor suppressor in renal cell carcinoma, were all confirmed as novel Per1 target genes. In an independent sample set, Cdkn1a, Cyclin E1, Claudin 1 and 12, and sFRP1 were all up‐regulated following Per1 knockdown in mpkCCDc14 cells. The chromophobe subtype of renal cell carcinoma is thought to arise from the collecting duct. Interestingly, dysregulation of the circadian clock genes Per2 and Cry2 was observed in patient chromophobe tumor samples compared to normal tissues. These observations support a role for the circadian clock in the control of cellular transformation in the kidney.
Ano1 as a regulator of proliferation
Authors: Stanich Jennifer E. , Gibbons Simon J. , Eisenman Seth T. , Bardsley Michael R. , Rock Jason R. , Harfe Brian D. , Ordog Tamas , Farrugia Gianrico
Published: June 2021

Ano1 is a Ca2+ activated Cl− channel expressed in several proliferating cell types including various epithelia, gastrointestinal stromal tumors, interstitial cells of Cajal (ICC) and CFPAC‐1 cells. The aim of this study was to determine if Ano1 regulates proliferation. ICC from Ano1(−/−) and Ano1(+/+) mice were cultured with Cl− channel inhibitors or in low Cl− media. CFPAC‐1 cells grown in low Cl− media were pulsed with EdU for 1 h. Incorporation was measured by flow cytometry. For immunoblots on mouse jejunum, antibodies to pRb, phospho(Ser773)‐pRb and GAPDH were used.

Fewer proliferating ICC were detected in Ano1(−/−) cultures (15 ± 3.5% Ki67+ ICC vs 25 ± 4% in Ano1(+/+), mean ± SEM, n = 4, p < 0.05). Cl− channel inhibitors DIDS (10 μM), niflumate (10 μM) and tamoxifen (1 μM) decreased ICC proliferation by 53, 53 and 59%, respectively (p < 0.05). This effect was reduced in cultures from Ano1(−/−) mice (29% and 19 % decrease, 22% increase, p > 0.05). Fewer proliferating ICC and CFPAC‐1 cells were detected in low Cl− media (% EdU+ ICC: 120 mM; 19.8 ± 5.3, 12 mM; 8.5 ± 3.2, n = 4, and % EdU+ CFPAC‐1: 120 mM; 29.5, 12 mM; 14.3 p < 0.05). Proportionally less Ser773‐phosphorylated pRb was detected in jejunum from Ano1(−/−) mice (Ano1(+/+); 5.98 ± 0.78, Ano1(−/−); 3.60 ± 0.49, mean ± SEM, n = 7, p < 0.05). In conclusion, Ano1 regulates proliferation possibly by promoting the G1/S transition of the cell cycle. Support: DK57061 and DK68055.
Detection of angiotensin II‐induced Ras activation in the trans‐Golgi network and the endoplasmic reticulum using BRET‐based biosensors
Authors: Hunyady Laszló , Soltész‐Katona Eszter , Erdélyi László , Várnai Péter , Balla András
Published: June 2021

Stimulation of the type I angiotensin receptor (AT1R), can lead to activation of small G proteins, such as Ras. In the present study, we investigated the effect of angiotensin II (AngII) on intracellular Ras activity in living HEK293 cells expressing AT1 receptors, using bioluminescence resonance energy transfer (BRET) biosensors. In order to investigate the subcellular localization of AngII‐induced Ras activation, we targeted the probes to various intracellular compartments, such as the TGN, the ER and early endosomes. We detected AngII‐induced Ras activation in the TGN and the ER, but not in endosomes. In cells expressing a tail‐deletion mutant AT1 receptor the AngII‐induced response was enhanced, suggesting that receptor internalization and arrestin binding are not required for AngII‐induced Ras activation in endomembranes. AngII was unable to stimulate Ras activity in the studied compartments in cells expressing the G protein coupling deficient mutant AT1 receptor. The presented data suggest that AngII can stimulate Ras activity in the TGN and the ER with a G protein dependently, which does not require arrestin‐binding, receptor internalization and EGF receptor transactivation.
Cell death in the atrioventricular canal myocardium determines ventricular activation patterns
Authors: Nanka Ondrej , Grim Milos , Sedmera David
Published: June 2021

We hypothesize that apoptosis in the AV myocardium influences maturation of atrioventricular conduction axis. We thus blocked apoptosis in chick embryonic heart and analyzed the effects of this treatment on cell death and ventricular activation patterns.

ED4 chick embryos were treated with the caspase inhibitor zVAD by intrapericardial injection. Spontaneously beating hearts isolated at ED8 were stained with voltage‐sensitive dye and imaged with a high‐speed camera. Amount of apoptotic cells was analyzed at ED 7 by whole mount LysoTracker Red or active caspase immunostaining and confocal microscopy. Hearts of embryos treated with zVAD showed a significantly increased proportion of immature (base to apex) activation patterns at ED8, including ventricular activation originating from right AV junction, a pattern never observed in control hearts. The number of apoptotic cells in AV canal myocardium was decreased at ED7 in the treated hearts. Apoptosis in the AV canal myocardium is thus an important contributor to activation pattern development. Its inhibition at critical stages can lead to persistence of accessory AV connections, which form a morphological substrate for ventricular pre‐excitation.

Supported by project of MSMT VZ 0021620806, LC 06061.
Sensitivity of Acute Myeloid Leukemia (AML) cells to a urokinase‐activated recombinant anthrax toxin
Authors: Abi‐Habib Ralph Joseph , Timsah Zahra , Darwich Manal , Alfano Randall , Liu ShiHui , Leppla Stephen , Frankel Arthur
Published: June 2021

We tested efficacy of a urokinase‐activated recombinant anthrax toxin (PrAgU2/FP59) on 11 human AML cell lines. PrAgU2/FP59 consists of PrAgU2, the cell binding domain in which the furin activation site is replaced by a urokinase plasminogen activator (uPA) activation site and FP59, the catalytic domain, consisting of Pseudomonas aeruginosa exotoxin A. PrAgU2 binds cells, is cleaved by uPA, heptamerizes, binds 3 molecules of FP59 and undergoes endocytosis releasing FP59 in the cytosol. FP59 ADP ribosylates EF2 causing protein synthesis inhibition and cell death. Introduction of the uPA activation site selectively targets this toxin to tumor cells expressing an active uPA system.

7/11 AML cell lines were highly sensitive to PrAgU2/FP59 (IC50 = 0.5–33 pM, % cell kill > 95%) while 4/11 cell lines were moderately sensitive to PrAgU2/FP59 (IC50 = 160–410 pM, % cell kill > 70%). Addition of pro‐uPA increased AML sensitivity to PrAgU2/FP59 by an average of 40 fold. Addition of a neutralizing anti‐uPA antibody decreased toxicity of PrAgU2/FP59 to AML cells by an average of 14 fold, demonstrating requirement for active uPA for selective activation of PrAgU2/FP59.

AML cells are sensitive to the cytotoxic effects of a urokinase‐targeted recombinant anthrax toxin with sensitivity depending on the expression of active uPA. PrAgU2/FP59 is a promising tumor selective therapeutic for the potential treatment of AML.
A Novel Inhibitor of the Histone Acetyl Transferase Activity of p300/CBP Associated Factor (PCAF)
Authors: Modak Rahul , Basha Jeelan , Kundu Tapas
Published: June 2021

We have identified a natural compound from an Indian traditional medicinal plant which is being used to treat fever, inflammatory diseases, and a variety of gastrointestinal ailments for thousands of years. The compound (MJTK‐1) has been shown to have antifertility, anthelmintic, antipyretic, analgesic, antiinflammatory and antibacterial activities. Here we report that MJTK‐1 inhibits lysine acetyl transferase (KAT) activity of Tip60, hGCN5, p300 and PCAF in vitro. MJTK‐1 specifically inhibits PCAF (KAT3B) up to 10μM concentration with Ki =1.2 ±0.5 μM. MJTK‐1 does not inhibit methyltransferases CARM‐1 and G9A. 25μM MJTK‐1 treated HEK 293T cells show specific down regulation of histone H3K9 acetylation without any alteration of other histone acetylation marks. Immunohistochemical and western blot analysis of liver tissues from MJTK‐1 treated mice showed specific down regulation of histone H3K9 acetylation. Presumably, MJTK‐1 at least with a great preference inhibits the HAT/ KAT activity of PCAF in vivo. Presently we are analysing the mechanism of this specificity as well as its biological implication in the context of epigenetics.

This work is supported by Indian Council for Agricultural Research and Jawaharlal Nehru Centre for Advanced Scientific Research, India.
Novel Cyclic Peptoid Ligands Targeting Oncoprotein Prothymosin α
Authors: Qi Xin , Kodadek Thomas
Published: June 2021

There is great interest in the discovery of synthetic molecules that recognize proteins with high specificity and affinity. Many cyclic peptide and depsipeptides have been found to display potent biological activities and often have better pharmacokinetic properties than their linear counterparts. We have applied a dual‐methionine strategy to incorporate a peptoid library into a cyclic scaffold to identify peptoid ligands targeting oncoprotein Prothymosin α that modulates apoptosis. To facilitate our screening effort, TentaGel beads that have bound the His‐ProT α were retained by the magnet through a peptoid—protein·anti‐His antibody·secondary antibody—Dynabead bridging interaction. We identified lead compounds that bind oncoprotein ProT α with moderate affinity. Furthermore, the microarray results confirmed that our cyclic peptoid hits target ProT with high specificity. In summary, we prepared a series of cyclic peptoid library and evaluated their potency for binding oncoprotein ProT. In particular, we demonstrated the advancement of using magnetic separation to accelerate the screening process and our approach allows more powerful selection of peptoid libraries.

This work was supported by the National Institutes of Health (1 P01‐DK58398).