bluuebunny/crossref_metadata_2025 huggingface dataset:
Development of the enteric nervous system and intestinal neuroendocrine systems in small for gestational age and average for gestational age piglets during the first month of life (1017.1)
Authors: Radlowski Emily , Le Boedec Kevin , Zhang Xuejin , Dilger Ryan , Johnson Rodney , Lezmi Stephane
Published: June 2021

In humans, fetal growth retardation is associated with an increased risk of intestinal problems (e.g. delayed meconium passage, abdominal distension, delay in tolerating enteral feeding, and necrotizing enterocolitis). These conditions may be related to deficits in gut development. A novel piglet model was developed to investigate the effects of birth weight and diet on the development of the enteric nervous system (ENS) and neuroendocrine system (serotonin, neurotensin, neuropeptide Y) via multivariate analyses. Newborn SGA or AGA piglets were randomized into two diet groups: sow reared (SR, 24h/day with the sow) or formula fed (FF, milk replacer) (n=4/group). Gut samples were collected at day (D) 1, 14 and 28, and used for immunohistochemistry. Along the study, innervation of the colon in SGA piglets was less developed compared to AGA piglets (p=0.007), except at D28. Serotonin and neuropeptide Y secreting cell numbers increased overtime with SR diet (p=0.0004 and p=0.004, respectively). The distribution of neurotensin cell numbers in different gut levels were highly variable overtime (p=0.0002). In conclusion, birth weight and diet influence ENS/neuroendocrine system development. This model may be useful to study intestinal maturation in low birth weight infants.
Aflatoxin M1 and ochratoxin A levels in breast milk in Ankara, Turkey (1017.8)
Authors: Barut Uyar Banugül , Karaağaoğlu Nilgün , Girgin Gözde , Gürbay Aylin , Karaağaoğlu Ergun
Published: June 2021

Aflatoxin and Ochratoxin known as mycotoxin are the highly toxic metabolic products of molds and one of the significant groups of chemical contaminants that humans are exposed. Exposure to mycotoxins is determined by the analysis of foods or biological fluids. Monitoring the levels of mycotoxin in biological fluids (urine, breast milk, umbilical cord blood, etc.) is accurate and reliable indicator of actual exposure. For this reason measuring contamination levels in breast milk is important. The objective of this study was to determine levels of aflatoxin M1 (AFM1) and ochratoxin A (OTA) in breast milk. Seventy samples of breast milk analyzed in this study were collected from randomly selected volunteer mothers gave birth in a maternity hospital in Ankara and are in lactation period for 20‐65 days. The determination of AFM1 and OTA levels were performed by enzyme‐linked immunosorbent assay (ELISA). AFM1 and OTA were determined in 9 (12.9%) and 34 (48.6%) samples of human milk, on average 5.73±0.74 ng/L and 0.14±0.03 ng/mL, respectively. In 7 mothers with AFM1 > detection limit (LD) OTA was also >LD. The results of the study indicate the contamination of human milk with mycotoxins. Consumption of safe food is important for public health especially for nutrition of mothers and infants who are within primary risk groups in public. Routine measurements of the toxin levels in foods and feeds should be carried out to prevent their harmful effects on health.
Relationship between baseline triglyceride concentration and triglyceride reduction with 4 g/d long‐chain omega‐3 acid ethyl esters (1035.6)
Authors: Nieman Kristin , Dicklin Mary , Bell Margie , Rains Tia , Maki Kevin
Published: June 2021

Long‐chain omega‐3 acid ethyl esters (OM3‐EE) reduce plasma triglycerides; this effect appears to be larger in individuals with hypertriglyceridemia vs. those with lower fasting triglycerides (TG). Limited information exists on the relationship between baseline TG and the TG response to OM3‐EE. This investigation assessed the relationship between baseline fasting TG concentration and % reduction in TG with use of 4 g/d of OM3‐EE (eicosapentaenoic or eicosapentaenoic + docosahexaenoic acid EE). Data from 16 published studies were assessed. Baseline mean/median TG concentrations ranged from 113 to 1203 mg/dL. Linear and non‐linear regression models were fit to the mean/median values with baseline and % TG change as the independent and dependent variables, respectively. Both weighted and non‐weighted models were fit and provided similar results, thus unweighted results are reported. The model fit improved slightly with loge transformation of the response data (r2 = 0.636 vs. 0.620). The best fit regression line was % TG change = 24.986 – 8.912 (ln baseline TG), p<0.001. Using this equation, predicted TG changes are ‐19.7, ‐25.8, and ‐32.0% with mean/median baseline TG concentrations of 150, 300, and 500 mg/dL, respectively. These findings confirm the relationship between baseline TG concentration and TG response, and provide a means for predicting response for clinical trial planning.
Comparison of endocannabinoid actions on metabolomic analysis of mouse and human myoblast cultures (1036.1)
Authors: Watkins Bruce , Kim Jeffrey , Pappan Kirk , Carlson Morgan
Published: June 2021

An aim of our laboratory is to characterize the actions of endocannabinoids (EC) and their precursor fatty acids on cannabinoid receptors in muscle. To understand the effect of the EC system (ECS) on myoblast metabolism an untargeted global metabolomic analysis was conducted. Primary mouse and human and C2C12 myoblasts were treated with 25µM docosahexaenoic acid (DHA), docosahexaenoylethanolamide (DHEA), arachidonoylethanolamide (AEA), 1µM of the cannabinoid receptor type 1 CB1 antagonist NESS0327, or BSA control. After 24h, cells were collected for metabolite analysis. In C2C12, mouse and human cultures, inclusion of BSA‐complexed DHA in culture media consistently led to higher DHA and 22:5n3 compared to the control. DHA treatment of C2C12 and mouse myoblasts also increased DHA containing lyso‐glycerol lipids and 1‐myristoylglycerol, 2‐myristoylglycerol, 1‐linoleoylglycerol, and 2‐linoleoylglycerol. In the mouse and human myoblasts, DHA treatment reduced choline phosphate but increased phosphoethanolamine and G3P compared to the control. Treatment of human myoblasts with cannabinoid agonist AEA and DHEA as well as NESS0327 led to higher amounts of glucose, lactate, and many intermediates of glycolysis compared to the control. These findings show that mouse and human myoblasts are responsive to DHA and ECS agonists/antagonist to influence glycolysis and lipid metabolism.
Antithrombotic activity of new 1‐ethylxanthine cyclohexylammonium salt (1054.5)
Authors: Samorodov Aleksandr , Kamilov Felix , Timirkhanova Galiya , Samorodova Albina , Khaliullin Ferkat , Murataev Daniyar
Published: June 2021

The previous findings in vitro show high antiaggregation activity of newly synthesized cyclohexilammonium salt of 2‐[1‐ethil‐3‐methyl‐7‐(dioxotiethanyl‐3)xantinyl‐8‐thio]acetic acid. The current findings are received in vivo on the model of collagen‐adrenaline thrombosis according to G.S. Di Minno subject to the FASEB Statement of Principles for the use of animals in research and education. Efficacy endpoint is the number of survived animals in comparison with the test group and comparator agents since the injection into the tail vein suspended matter of collagen and adrenalin upto the moment of death or during 14 days of monitoring. The survival analysis was carried out with the help of Kaplan‐Meier method. The difference in survival between the groups was assessed with the help of Wilkonson criterion.

Intraperinoneal injection of the new secondary 1‐ethylxantin reduced mortality in lab mice by 1,3 times (P1<0,01) in comparison with the camparator agents. Thus, cyclohexilammonium salt of 2‐[1‐ethil‐3‐methyl‐7‐(dioxotiethanyl‐3)xantinyl‐8‐thio]acetic acid is a potential antithrombotic agent.
Astrocyte and microglial activation by insulin/saline treatment of diabetic ketoacidosis (1072.8)
Authors: Lo Weei , O'Donnell Martha , Glaser Nicole
Published: June 2021

Diabetic Ketoacidosis (DKA) and/or its treatment with insulin and saline (I/S) may cause brain injury in children. The mechanism of DKA‐related brain injury is poorly understood. It is unclear whether injury is mainly caused by DKA per se or I/S treatment. Similarly, it is unclear whether some brain regions are more susceptible than others to injury. We used immunohistochemistry (IHC) in a rat DKA model to assess markers of brain injury. Juvenile rats with streptozotocin‐induced DKA were treated with I/S until DKA resolution. IHC of brains during DKA and 4, 24, and 72 hr after I/S were examined for glial fibrillary acidic protein (GFAP) and CD‐11b protein (OX42) to evaluate changes in astrocytes and microglia in the hippocampus, cortex and striatum. GFAP and OX42 expression was increased in the hippocampus of DKA rats. GFAP staining intensity peaked 4 hours after beginning I/S treatment (34% increase). OX42 expression was elevated at 4 hr and through 24 and 72 hr (26, 26 and 21% increases). OX42, but not GFAP, was also significantly increased in cortex at 4, 24, and 72 hr after I/S treatment (28, 21 and 23% increases). Neither GFAP nor OX42 were changed in the striatum. Our data suggest that reactive gliosis and microglial activation occur during DKA‐related brain injury. Reactive gliosis is most prominent during I/S treatment, but microglial activation persists up to 72 hr, suggesting a sustained response to injury.

Grant Funding Source: Supported by ADA
Changes in IgG sialylation and glycosylation in pulmonary arterial hypertension (1089.3)
Authors: Morrow Rebekah , Cioffi Eugene , Murphy Frank , Cioffi Donna
Published: June 2021

Rationale: Anti‐endothelial cell antibodies are found in both idiopathic and connective tissue disease associated pulmonary arterial hypertension (PAH). IgG is the most abundant immunoglobulin and is heavily glycosylated. Sialic acid is a monosaccharide often found on the end of glycan chains, and sialylation of IgG can alter IgG binding to its receptors. Changes in IgG sialylation and N‐linked glycan composition occur in numerous diseases, but it is unknown whether changes occur in PAH. We sought to determine whether IgG sialylation and glycosylation are altered in PAH patients, and if these changes affect IgG binding to endothelial cells (EC). Methods: We purified IgG from healthy volunteers (HV), idiopathic PAH (IPAH), or connective tissue disease‐associated PAH (CTD) patients. Lectin blots were used to analyze IgG sialic acid linkages and glycosylation residues. IgG was treated with sialidases to remove sialic acids or glycosidases to remove N‐linked glycans. The cleaved glycans were analyzed by ESI mass spectrometry. Both untreated IgG and sialidase‐treated IgG were used in a cell based ELISA to examine the effect of sialylation on IgG binding to EC. Results: Lectin blots and mass spec show marked differences between HV and PAH IgG. PAH IgG treated with sialidase has decreased binding to EC, whilst untreated PAH IgG has increased binding to EC as compared to HV IgG. These data suggest that sialylation and glycosylation differences in IgG from PAH patients affect binding to EC.

Grant Funding Source: Supported by R01HL107778
Impact of nitrate supplementation via beetroot juice on capillary hemodynamics in skeletal muscle of rats in chronic heart failure (1106.16)
Authors: Ferguson Scott , Holdsworth Clark , Wright Jennifer , Fees Alexander , Musch Timothy , Poole David
Published: June 2021

Chronic heart failure (CHF) causes deficiencies in skeletal muscle blood flow resulting in compromised skeletal muscle capillary red blood cell (RBC) distribution and hemodynamics. These impairments are thought to be due, in part, to reductions in nitric oxide (NO) bioavailability within skeletal muscle. In healthy animals, nitrate (NO3‐) supplementation via beetroot juice (BR) elevates skeletal muscle blood flow and raises the pressure head for capillary‐myocyte O2 flux during exercise presumably following a stepwise reduction to NO in vivo. We tested the hypothesis that BR supplementation would increase the % of capillaries supporting RBC flow at rest and during contractions in CHF rats. CHF was induced in young adult male Sprague‐Dawley rats via myocardial infarction (MI). Following a 5‐week recovery period rats were given BR ([NO3‐] 1 mmol/kg/day, CHF+BR) or water (CHF) for 5 days. MI size was not different between groups (CHF: 28 ± 5, CHF+BR: 28 ± 6 %). Intravital microscopy was used to study the in vivo spinotrapezius muscle microcirculation at rest and during 180 s 1Hz twitch contractions (6‐8 V). The percentage of capillaries supporting continuous RBC flow was elevated in CHF+BR rats at rest (CHF: 65 ± 6, CHF+BR: 76 ± 2 %, P蠄0.05) and during contractions (CHF: 78 ± 6, CHF+BR 82: ± 2 %, P<0.05). The improvements seen herein are likely due to enhanced arteriolar vasodilation mediated, in part, by elevated NO bioavailability. These results have important implications and suggest that NO3‐ supplementation via BR may constitute a viable therapeutic modality that improves muscle vascular and potentially metabolic function in CHF.

Grant Funding Source: Supported by NIH‐HL108328
Dietary supplementation with polyunsaturated fatty acid during pregnancy modulates DNA methylation at IGF2/H19 imprinted genes and growth of infants (1120.3)
Authors: Romieu Isabelle , Lee Hosun , Barraza Albino , Biessy Carine , Duarte‐Salles Talita , Sly Peter , Ramakrishnan Usha , Rivera Dommarco Juan , Herceg Zdenko
Published: June 2021

Epigenetic regulations of imprinted genes are highly plausible explanation for the associations of dietary exposures in early life and the onset of diseases in adulthood. We tested whether prenatal dietary supplementation with docosahexaenoic acid (DHA) during pregnancy may modulate epigenetic states at birth in a randomized intervention trial among Mexican pregnant women supplemented daily with 400 mg of DHA or a placebo from 18 to 22 week of gestation to parturition. We applied quantitative profiling of DNA methylation states at IGF2 promoter 3 (IGF2 P3), IGF2 DMR and H19 DMR in cord blood mononuclear cells of the DHA‐supplemented group (n=131) and the control group (n=130).There were no significant differences between the DHA and the control groups for DNA methylation levels at imprinted genes; however in stratified analyses, we observed a positive association between DNA methylation levels and maternal BMI; IGF2 DMR methylation was higher in the DHA than the control groups in infants from overweight mother (p=0.03) and H19 DMR, methylation levels were significantly lower in the DHA than control groups in infants from normal weight mothers (p=0.01). Prenatal DHA supplementation modulated IGF2/H19 methylation in infants and methylation levels at IGF2/H19 imprinted regions were related with maternal BMI. Epigenetic mechanism may be modulated by DHA with potential impacts on growth and development of children.

Grant Funding Source: NIH‐NICHHD (RO1HD058818), NIH (RO1HDO43099) and CONACYT Mexico (SALUD‐2008‐01‐87121)
Evidence of impaired vasodilation during exercise in heart failure with preserved ejection fraction (1156.3)
Authors: Lee Joshua , Barrett‐O’Keefe Zachary , Nelson Ashley , Ryan John , Nativi Jose , Richardson Russell , Wray D
Published: June 2021

Exercise intolerance is a hallmark characteristic of heart failure (HF), which may be related to impaired exercise‐induced vasodilation in the exercising muscle vasculature. While reductions in exercising limb blood flow have been observed in patients with systolic HF no studies, to date, have evaluated vasodilation in HF patients with preserved ejection fraction (HFpEF) during exercise. Thus, in six HFpEF patients (68±8 yrs) and six age‐ and sex‐matched controls (69±9 yrs), arterial pressure (Finapres) and leg blood flow (LBF; ultrasound Doppler) were assessed during knee‐extension exercise at 0, 5, 10, and 15 W. Resting arterial pressure (HFpEF: 98±2 mmHg; controls: 105±16 mmHg) and LBF (HFpEF: 276±57 mL/min; controls: 299±59 mL/min) were similar between groups. Across work rates, arterial pressure was similar between groups, yet LBF was 30‐40% lower in HFpEF compared to controls (1,256±219 vs. 1,791±151, 1,420±206 vs. 2,102±179, 1,666±91 vs. 2,459±417, and 1,745±206 vs. 2,874±206 mL/min, HFpEF vs. controls at 0, 5, 10, and 15 W, respectively). These data indicate that exercise‐induced vasodilation is markedly impaired in HFpEF patients, which likely contributes to exercise intolerance in this patient population.

Grant Funding Source: Supported by NIH PO1 HL‐091830 (R.S.R.) VA RR&D E6910R (R.S.R.) AHA 0835209N (D.W.W)
Osteoderm accretion as proxy for whole body growth in the American alligator (1161.3)
Authors: Vasconcellos David , Owerkowicz Tomasz , Eme John , Blank Jason , Elsey Ruth , Hicks Jim
Published: June 2021

Osteoderms are small bones found in the dermal layer of sauropsid scales, form an integral part of integumental armour and may participate in thermoregulation. Whether growth of osteoderms parallels that of the whole animal has not yet been ascertained. We studied histomorphometry of cervical osteoderms of juvenile female American alligator (Alligator mississippiensis) reared at 30°C on ad libitum diet. All animals had undergone a cardiac surgical procedure, which either rendered their circulatory system in‐series (experimental n=24) or retained the original in‐parallel design (sham n=36). In addition, animals were assigned to one of three exercise groups: sedentary, running or swimming (n=20 each) for 18 months. Body mass and length measurements were taken every other week, and animals received injections of fluorescent dyes (calcein, alizarin) during the growth period. We found a positive correlation (ANCOVA, p<0.01) between periosteal bone deposition in osteoderms and body growth. Neither surgical manipulation nor exercise had a statistically significant effect on this relationship. We propose annular growth lines in osteoderms can be used as part of non‐lethal methodology to track growth in populations of wild crocodilians. Further, osteoderm histology can inform our understanding of growth trajectories in extinct archosaurs, regardless of their cardiovascular design or activity patterns.
Acute slow‐paced breathing increases periods of sympathetic nervous system quiescence (1170.12)
Authors: Mozer Michael , Fadel Paul , Johnson Christopher , Wallin B , Charkoudian N , Drobish Justin , Joyner Michael , Wehrwein Erica
Published: June 2021

Slow‐paced breathing is a simple strategy to reduce psychological stress. We tested the hypothesis that slow paced breathing would decrease muscle sympathetic nerve activity (MSNA). Ten young, healthy males were studied. The subjects were instructed to rest supine and breathe normally (baseline) or at six breaths per minute, each for 20 minutes. MSNA was measured (peroneal microneurography) and analyzed 5 minutes into each stage for a steady state measurement. Statistical analysis was completed using one way RM ANOVA and Tukey’s posthoc analysis (reported mean±SEM). Average respiratory rate was 11.9 ± 1.1 breaths/min during baseline and 5.8 ± 0.7 breaths/min during slow‐paced breathing. There was no significant difference from baseline compared to slow paced breathing in MSNA burst frequency (18±2.0 vs 18±2.40 bursts/min), burst incidence (31.2±4.01 vs. 30.4±3.80 bursts/100 heartbeats) or burst area (12.70±2.18 vs 13.02±1.98%). However, there was a change in the pattern of burst firing with a significant increase in quiescent periods between burst clusters during slow paced breathing compared to baseline (6.50±0.61 vs. 3.59±0.41 seconds), which was accompanied by a significant decrease in the number of bursts/cluster (2.9±0.38 vs. 5.6±0.8 bursts). This has physiological relevance since there was a significant decrease in plasma norepinephrine, increase in blood flow, and a reduction in mean arterial pressure during these same periods. These findings indicate that despite no change in burst incidence, slow paced breathing alters the pattern of MSNA firing, which may contribute to favorable cardiovascular changes.

Grant Funding Source: Supported by NIH HL‐083947
Hyperglycemia increases prorenin receptor localization at the cell plasma membrane (1173.7)
Authors: Prieto Minolfa , Arita Danielle , Bourgeois Camille , Satou Ryosuke
Published: June 2021

The prorenin receptor (PRR) elicits intracellular signals linked to tissue fibrosis based upon its capability to bind prorenin at the plasma membrane (PM) which may contribute to nephropathy and microvascular changes in diabetes mellitus (DM). To test the hypothesis that hyperglycemia increases the capability of PRR to bind prorenin by increasing membrane bound PRR to PM in collecting duct cells, we used type 1 DM rats (N=10) induced with a single STZ injection (ip; 200ng/kg) as well as in M‐1 cells to assess PRR trafficking alteration induced by hyperglycemia. After 7‐days induction, STZ‐rats had levels of plasma glucose of 428±13 vs. 138±9 gr/dL and plasma insulin 0.07±0.02 vs. 2.42 ng/mL compared to control rats. By immunoflurescence (IF) studies of the rat kidney section, PRR was mainly localized predominantely on the pical aspect of collecting duct cells in STZ‐rats while in control rats most of PRR immunoreactivity was observed intracellularly. In methanol‐fixed M‐1 cells, PRR was localized mainly to the perinuclear areas in normal glucose (NG; 1g/L glucose + 1 g/L mannitol)‐treated cells. However, in high glucose (HG; 4g/L)‐treated cells, PRR was predominantly localized toward the surface, as it was appreciated in the de‐convoluted images. Although PRR mRNA levels were not different in the collecting ducts of renal medulla between groups; PRR protein levels (37 kDa) were significantly higher in PM extracts from M‐1 cells treated with HG for 5 minutes, 1, and 6 hours (with a maximum peak observed at 1 hour) than in those treated with NG. These data suggest that hyperglycemia induces PRR trafficking alterations of PRR. The effect of hyperglycemia to increase PRR abundance in PM in the collecting duct may be a novel mechanism underlying the increased hypertension seen in the diabetic patients.

Grant Funding Source: Supported by NIH‐DSNTR grant
Children’s docosahexaenoic acid (DHA), but not maternal DHA in pregnancy, is associated with psychometric tests scores at 5‐6 years of age (124.2)
Authors: Mulder Kelly , Innis Sheila , Richardson Kelly
Published: June 2021

Docosahexaenoic acid (DHA) is an important component of neural lipids, affecting neural function throughout life. However, the dietary, genetic and other variables that impact DHA transfer to the brain, and the potential for early deficiency to have lasting effects remains unclear. We addressed the effect of DHA in gestation and children’s intake and blood lipid DHA on cognition at 5.75 years of age. Pregnant women, n=271 were enrolled at 16 weeks gestation and randomized to 400 mg DHA/day or placebo until infant delivery; 95 children returned for follow up, and were assessed with 153 additional children. Diet was assessed using 3 day records and frequency questionnaires, and venous blood was collected. Cognitive tests included the Peabody Picture Vocabulary Test (PPVT) and Kaufman Assessment Battery for Children (KABC). The median (IQR) DHA intake was 49.4 (23.8‐102) mg/day. The red blood cell mean ± SD DHA was 5.14 ± 1.50 percent total fatty acids. No evidence of persisting effects of DHA in pregnancy on child development were found. Children’s own DHA status, but not DHA intake, was associated with better PPVT and KABC Sequential, Learning, Simultaneous, and Mental Performance scores (P <0.05). Child blood lipid 22:5n‐6 and the 22:5n‐6/DHA ratio were associated with poorer PPVT and KABC performance. DHA intake explained only 15% of variability in RBC DHA, raising complex questions on the relationship between diet, DHA transfer to membrane lipids, and neural function.
Risk factors for prelacteal feedings in seven Latin America and Caribbean countries: a multilevel analysis (131.7)
Authors: Boccolini Cristiano , Pérez‐Escamilla Rafael
Published: June 2021

Objectives: Identify risk factors for prelacteal feedings (i.e., offering liquids before milk begins to flow) in the first three days of life in Latin America and the Caribbean (LAC).

Methods: The study used secondary data from Demographic and Health Surveys conducted in seven countries (Colombia/2010, Bolivia/2008, Guiana/2009, Haiti/2005, Honduras/2005, Peru/2007 and Dominican Republic/2007). Binomial regression, with random effects at region and country level, tested the influence of potential risk factors on the likelihood of prelacteal feeding. Countries’ data sets were pooled (N=58,056 children).

Results: In the first three days of life, mothers gave infant formula (17.9%), tea/infusions (6.4%), milk other than breast milk (5%), and/or other liquids (4.5%) before breast milk began to flow. Prelacteal feeding’s prevalence varied from 17.6% in Guiana to 55% in Dominican Republic. Independent risk factors for prelacteal feeding were: wealth (1.33, 1.24‐1.46), higher maternal educational level (1.46, 1.32‐1.60), cesarean delivery (2.34, 2.28‐2.41). Hospital delivery (vs. home) was associated with a lower likelihood of prelacteal feeding (0.72, 0.64‐0.80).

Conclusion: Findings suggest that the prevalence of prelacteal feeding across countries in LAC varies substantially and that socio‐economic and biocultural factors explain some of the variance in prelacteal feeding within LAC countries.

Grant Funding Source: Supported by Brazil's National Council for Scientific and Technological Development (CNPq)
Financial costs of Mama‐SASHA ő a project to improve health and nutrition through an integrated orange flesh sweet potato production and health service delivery model (132.6)
Authors: Self Julie , Kedera Ellah , Grant Frederick , Wamalwa Moses , Hu Jia , Low Jan , Cole Donald , Webb Girard Amy , Levin Carol
Published: June 2021

Mama‐SASHA project aims to improve the health status of pregnant women and nutritional status of children up to two years through an integrated orange‐flesh sweet potato (OFSP) and health service delivery strategy in Western Kenya. Nutrition education and vouchers for OFSP vines are provided during antenatal care visits, with additional nutrition education and support provided in the communities and through pregnant mothers clubs (PMCs). The purpose of this study is to analyze the financial costs of the Mama‐SASHA project. We use a microcosting approach based on project expense reports to estimate financial costs during the period 2011‐2013, allocating costs by activity and inputs by implementing organization. Project monitoring data were used to estimate project output and number of beneficiaries reached. Financial costs were incurred by two agricultural NGOs and one health NGO, totaling $344,860 (USD). Over 5,400 women participated in monthly PMCs at a cost of $63 per woman. Of 4,629 women who received vouchers, 3,281 women redeemed vouchers and planted OFSP at a cost of $105 per beneficiary. There are limited comparable cost estimates in the literature; however, this estimate falls in the range of published cost estimates for community‐based therapeutic feeding programs supported by community health workers with some referrals to health centers.
Delaying and reducing medication use as a coping mechanism for food insecurity (268.6)
Authors: Herman Dena , Afulani Patience , Harrison Gail , Coleman‐Jensen Alisha
Published: June 2021

Objective: To determine if food insecure households are more likely to skip, delay or reduce medications to compensate for lack of household resources to purchase food. Methods: We used data from a combined sample of the 2011 and 2012 National Health Interview Survey (n=52,699). We conducted bivariate and logistic regression analyses to examine the association between food security (FS) and medication use; controlling for demographics, lifestyle, BMI, chronic disease, disability, health care coverage and self‐rated health. Sample weights were applied to account for the complex sample design. Results: Seventy eight percent of respondents reported high FS, 8% marginal FS, 6.9% low FS and 6.0% very low FS. Seven percent reported skipping medications; 7.6% used fewer medications; 9.4% delayed filling a prescription; and 20% lowered the dose of medication. Net of other factors the odds of skipping medications increased with increasing severity of food insecurity (OR of 1.67 for marginal FS, 2.86 for low FS and 4.14 for very low FS). The results were similar for delaying or reducing medications. Also women, low income, working part‐time, current smokers, severely obese, no health care coverage, having a chronic disease, functional limitation and poor self‐rated health had higher odds of skipping, delaying or reducing medications. Conclusion: Delayed or reduced medication use may be used as a coping mechanism among the food insecure and could lead to further negative health consequences for those who may already be suffering from chronic disease conditions.

Grant Funding Source: Supported by a cooperative agreement with the ERS, Contract#: 58‐4000‐3‐0009
Fasting rapidly increases fatty acid oxidation in white adipose tissue (269.2)
Authors: Torchon Emmanuelle , Hulver Matthew , McMillan Ryan , Voy Brynn
Published: June 2021

Upregulating fatty acid oxidation in white adipose tissue in mice protects against diet‐induced obesity and excess plasma NEFA levels. At least part of this capacity results from the induction of brown‐like adipocytes within classical white depots, making it difficult to determine if white adipocytes contribute. Avian genomes lack a gene for uncoupling protein 1 and are devoid of brown adipocytes, making them a useful model in which to study lipid metabolism in white adipocytes. We recently reported that a brief (5 hr) period of fasting upregulated expression of genes involved in mitochondrial and peroxisomal fatty acid oxidation in white adipose tissue of broiler chickens. The objective of this study was to determine if the effects on gene expression manifested in increased rates of fatty acid oxidation. Abdominal adipose tissue was collected from 21 day‐old broiler chicks that fasted for 3.5, 5 or 7 hrs, or fed ad libitum. Fatty acid oxidation was determined by measuring and summing 14CO2 production and 14C‐labeled acid‐soluble metabolites from the oxidation of [1‐14C]palmitic acid. Citrate synthase activity was measured spectrophotometrically. Fasting induced a progressive increase in complete oxidation which was significantly different from controls in the 5.5 hr (p=0.0037) and 7 hr (p=0.0021) groups (1.14, 1.2, 1.49 and 1.95 nmol/mg protein/hr; control, 3.5, 5 and 7 hrs, respectively). Citrate synthase activity increased significantly after 7 hrs of fasting. Fasting did not significantly alter the production of acid soluble metabolites, an index of incomplete fatty acid oxidation. These results confirm that fasting rapidly increases fatty acid oxidation in white adipose tissue by upregulating the transcription of key regulatory enzymes and proteins. Identifying the underlying mechanism may provide new therapeutic targets through which to increase fatty acid oxidation in situ and protect against obesity and the detrimental effects of excess NEFA on adipocyte insulin sensitivity.
Biofortified orange maize is as efficacious as a vitamin A supplement in Zambian children even on the background of high liver reserves of vitamin A (39.5)
Authors: Gannon Bryan , Kaliwile Chisela , Arscott Sara , Schmaelzle Samantha , Masi Cassim , Chileshe Justin , Tanumihardjo Sherry
Published: June 2021

Biofortification is an emerging strategy to relieve vitamin A (VA) deficiency. Biofortified (orange) maize contains β‐carotene (βC) and has shown efficacy for VA status in animal models, but has yet to be confirmed in humans. A randomized, placebo‐controlled orange maize efficacy trial was conducted in rural Zambia. The paired 13C‐retinol isotope dilution test, the most accurate and sensitive indirect marker of vitamin A status, was used to measure total body reserves (TBR) of vitamin A before and after a 90‐d intervention period. Treatment arms were white maize with placebo oil (VA‐), white maize with VA oil (VA+), and orange maize with placebo oil (orange). Change in TBR showed a treatment effect (P = 0.011) with VA+ being significantly greater than VA‐, while orange was statistically similar to both (203 ± 386, ‐4.50 ± 208, 109 ± 341 µmol, respectively). However, data were not normally distributed (P < 0.0001), medians were VA‐ 12.6, VA+ 97.8, orange 83.7 µmol, and non‐parametric analysis showed VA+ and orange were statistically similar and different from VA‐. Baseline estimates of liver concentration were 1.13 ± 0.41 µmol/g, with 59% > 1 µmol/g, the current sub‐toxicity cutoff. None were < 0.1 µmol/g. The bioconversion factor was calculated to be 10.4 µg βC:µg retinol using the middle three quintiles of change in TBR from each treatment group. Change in serum retinol showed no treatment effect (P = 0.16). The βC in maize is a very effective vitamin A source when consumed as a staple food and could avoid potential toxicity issues with preformed VA from supplementation or fortification as seen in this population. Further use of stable isotopes should be used to elucidate the VA status of various populations, and serum retinol should be used with caution in setting global health policy.

Grant Funding Source: Supported by HarvestPlus and NIH grant T32‐DK007665 (BG)
A small diameter bioactive prosthetic vascular graft with activated protein C (546.9)
Authors: Linderman Shannon , Araya Joel , Pathan Saif , Nelson David , Phaneuf Matthew , Contreras Mauricio
Published: June 2021

To date, there are no small (less than 5 mm) internal diameter vascular grafts that are FDA approved for clinical use due to high failure rates from thrombosis and increased cell proliferation, conditions that may be improved with use of enhanced bioengineered graft materials. Here we show that long term graft viability and patency can be improved by covalently binding activated protein C (APC) to prosthetic vascular graft surfaces due to APC’s inherent physical properties. Electrospun polyester (Dacron) grafts covalently bound with APC and woven plain Dacron prosthetic graft controls were evaluated in‐vivo in a common carotid artery bypass canine model with and without Plavix treatment. Blood work samples were taken from each canine (n=12) at 14, 30, 44 or 60 days after graft implantation. Explanted grafts were examined macroscopically and histologically. Upon comparison of all time points, APC grafts offered 25‐50% improvement in patency compared to control grafts. Plavix treatment increased patency of APC grafts from 50% to 100% over 60 days, while the control grafts showed no significant improvement in patency with Plavix treatment. Furthermore, histological evaluation showed that APC grafts had partial endothelial cell (EC) coverage of the luminal surface (CD31 positive) in the midd‐portion while the Dacron control grafts did not. At the anastomosis, there were no distinct differences in overall graft healing (SMC‐a, Ki67). Our results suggest that Dacron‐APC grafts not only offer a significant improvement in patency over woven control grafts, but also promote EC healing throughout the luminal surface of the graft. Perhaps longer term studies are needed to asses if partial EC coverage could become complete over time.

Grant Funding Source: NIH‐803IR Phase II Grant.
Synthesis of fatty acid phytyl esters in chloroplasts of Arabidopsis thaliana (617.7)
Authors: Wehler Regina , vom Dorp Katharina , Doermann Peter
Published: June 2021

During senescence or stress, chloroplast membranes undergo severe changes. Galactolipids are broken down, releasing free fatty acids. At the same time, the plastoglobule size increases. Plastoglobules serve as lipid‐storage particles in chloropalsts, containing mostly neutral lipids such as triacylglycerol, tocopherol and fatty acid phytyl esters. Two genes (Phytyl Ester Sythase 1, PES1; Phytyl Ester Synthase 2, PES2) were identified that are primarily responsible for the synthesis of fatty acid phytyl esters in chloroplasts. In addition to phytyl ester synthesis, PES1 and PES2 harbor diacylglycerol acyltransferase activity and are presumably involved in the synthesis of triacylglycerol. An Arabidopsis double mutant (pes1 pes2) deficient in the two PES genes shows a reduction in the amounts of fatty acid phytyl esters and of triacylglycerol by more than 90 and by 30%, respectively. It is known that the bulk of triacylglycerol is produced by cytosolic acyltransferases at the ER. To investigate the role of PES1/PES2 in plastidial triacylglycerol production, we isolated chloroplasts from wild type and pes1 pes2 plants and analyzed the lipid composition of the individual chloroplast fractions. In addition to PES1 and PES2, the Arabidopsis genome contains 4 other genes encoding Esterase‐Lipase‐Thioesterase (ELT) proteins: ELT3, 4, 5 and 6, all of which are predicted to be chloroplast‐localized. We obtained insertion lines for ELT3, 4, 5 and 6 and analyzed the homozygous plants for triacylglycerol and fatty acid phytyl ester content.

Grant Funding Source: Deutsche Forschungsgemeinschaft
Naringenin, a major bioactive lead of Thymus vulgaris, inhibits growth, induces apoptosis and modulates cell cycle and apoptosis‐regulatory gene expression in human cancer cells (657.18)
Authors: Abaza Mohamed Salah , Orabi Khaled , Al‐Attiyah Raja'a , Al‐Qattan Ebtehal
Published: June 2021

In the present study Thymus vulgaris was extracted in 96% ethanol then subjected to solvent/solvent partitioning processes and purification techniques. The identities of the isolated compounds, naringenin (Nar) and aromadendrin (Aro) were identified using different spectral techniques. Nar inhibited the growth of human colorectal, breast, and melanoma cancer cell lines in a dose‐ and time‐dependent manner, arrested their growth in S‐phase and G2/M‐phase or S‐phase and potently induced their apoptosis. Nar differentially down‐regulated the expression of Cdk4, Cdk6, Cdk7, Bcl2, x‐IAP and c‐IAP‐2, differentially up‐regulated the expression of p18, p19, p21, caspases 3, 7, 8 and 9, Bak, AIF and Bax in both human colorectal and breast cancer cell lines. Nar could be a potential agent for prevention and/or treatment of human cancers.

Grant Funding Source: this work was supported by Kuwait University, Research Grant No.[YS06/10]
Glutathione levels and susceptibility to chemically induced injury in human prostate cancer cell lines (663.11)
Authors: Lash Lawrence , Putt David , Jankovich Adam
Published: June 2021

More aggressive or invasive prostate cancer cells (PCCs) are often resistant to chemotherapy and have poor prognosis. Two immortalized human PCC lines, PC‐3 (more aggressive) and LNCaP (less aggressive) cells, were compared with regard to cellular redox state and susceptibility to either oxidants or glutathione (GSH) depletors. We tested the hypothesis that compared to less aggressive PCCs, more aggressive PCCs exhibit higher GSH concentrations and are relatively resistant to cytotoxicity from oxidants. PC‐3 cells exhibited 4.2‐fold higher GSH concentration than LNCaP cells but only exhibited lower lactate dehydrogenase (LDH) release after toxicant exposures at some time points. Only LNCaP cells underwent diamide‐induced apoptosis. Expression of several proteins involved in stress response and regulation of cell growth and proliferation were measured to determine their possible role in redox state and susceptibility to toxicants. PC‐3 cells exhibited higher levels of pro‐apoptotic and lower levels of anti‐apoptotic proteins than LNCaP cells. Consistent with a compensatory response to oxidative stress, LNCaP cells exhibited higher levels of certain antioxidant and stress response proteins than PC‐3 cells. Thus, significant differences in redox status and expression of proteins involved in apoptosis and stress response may contribute to PCC aggressiveness.

Grant Funding Source: None.
Leptin acutely inhibits respiratory function in neonatal rats (712.18)
Authors: Wollman Lila , Powell Gregory , Levine Richard , Fregosi Ralph
Published: June 2021

Leptin resistance is associated with impaired respiratory function, obesity related respiratory disorders, and a higher incidence of respiratory illness in children of obese mothers. There have been few attempts to identify its site of action in respiratory centers of the brain and no studies on its respiratory effects in neonates. We hypothesize that the direct effect of leptin on the respiratory system is inhibitory, as it is in other brain regions, but that effects on metabolism may override this inhibition in the whole animal. We performed experiments on neonatal rat pups, postnatal day 0 to 6. Bath application of 300nM leptin to four brainstem spinal cord (BSSC) preparations had varied effects on burst amplitude and frequency of Cervical 4 ventral roots (C4VR). In three split bath BSSC preparations, 100uL of 0.5mg/ml leptin applied to the medullary compartment slowed or abolished C4VR rhythm, which recovered with washout. Whole bath application of 300nM leptin to five rhythmic medullary slices abolished hypoglossal rootlet rhythm, which recovered with washout. Microinjections of 0.5 mg/ml leptin, but not vehicle injection, into the pre‐Botzinger complex of two BSSC preps slowed or abolished C4VR rhythm. These preliminary results suggest that leptin receptors are located on pre‐Botzinger complex cells, and that leptin’s action is inhibitory.

Grant Funding Source: Supported by ROIHD071302
Vitamin A and iron status of women and children in urban Cameroon before and after introduction of vitamin A‐fortified vegetable oil and iron‐fortified wheat flour (804.18)
Authors: Engle‐Stone Reina , Ndjebayi Alex , Nankap Martin , Gimou Marie‐Madeleine , Friedman Avital , Brown Kenneth
Published: June 2021

Few data are available on the effectiveness of mass food fortification programs. We conducted representative surveys 2 y before and 1 y after introduction of vitamin A (VA)‐fortified cooking oil and iron‐fortified wheat flour to assess program impact on VA and iron status in urban Cameroon. In each survey, 10 different households were selected within each of the same 30 clusters (n=~300/survey). Malaria and plasma indicators of inflammation (CRP, AGP) and VA (retinol‐binding protein, pRBP) and iron (ferritin, pF; soluble transferrin receptor, sTfR) status were assessed among women 15‐49 y and children 12‐59 mo. Staple food intake was measured by FFQ, and post‐fortification oil and flour samples were collected. Pre‐fortification, low inflammation‐adjusted pRBP was common among children (33% < 0.83 µmol/L), but not women (2% < 0.78 µmol/L); > 20% of women and children had low adjusted pF. Oil and flour were each consumed by >80% of participants in the past week. Post‐fortification, 44% of oil and 76% of flour samples were fortified. Controlling for inflammation and malaria, there was no change in pRBP among women (1.41 to 1.40 µmol/L) or children (0.87 to 0.88 µmol/L), but pF increased (women: 37 to 47 µg/L; children: 39 to 51 µg/L, both P < 0.05) and sTfR decreased (7.7 to 6.2 and 10.6 to 8.2 mg/L, P < 0.01). Plausibility analyses will determine whether the observed changes are attributable to the fortification program.

Grant Funding Source: Supported by the Michael and Susan Dell Foundation, Sight and Life, and UNICEF
Bioavailability of enteric‐coated microencapsulated calcium during pregnancy: a randomized crossover trial in Bangladesh (804.4)
Authors: Pezzack Brendon , Zlotkin Stanley , Abrams Steven , Hawthorne Keli , Al Mahmud Abdullah , Baxter Jo‐Anna , Aimone Phillips Ashley , Islam Munirul , Dimitris Michelle , Ahmed Tahmeed , Roth Daniel
Published: June 2021

Prenatal calcium (Ca) and iron (Fe) supplements are recommended in settings of low dietary Ca intake and high prevalence of anemia. However, concurrent Ca and Fe administration may inhibit Fe absorption. Therefore, we developed a multi‐micronutrient powder containing Fe (60 mg), folic acid (400 µg), and Ca (0.5, 1.0 or 1.5 g) in which calcium carbonate granules were microencapsulated with a pH‐sensitive enteric coating to delay intestinal release and limit Ca‐Fe interactions. To compare the fractional intestinal absorption (fAb) of Ca from enteric‐coated (EC) Ca granules versus uncoated (non‐EC) granules, we conducted a randomized crossover trial among pregnant women (n=49) in Dhaka. fAb was estimated by a dual stable isotope method (44Ca‐labeled granules and intravenous 42Ca), based on the relative recovery of 44Ca vs 42Ca in urine over 48 hours. Mean (±SD) fAb from EC Ca was significantly less than from non‐EC Ca (2.7±2.2 % vs. 16.5±10.0 %; p<0.0001) at all 3 Ca doses. In conclusion, the pH‐sensitive enteric coating substantially reduced Ca absorption. Therefore, in its current formulation this novel enteric‐coated Ca‐Fe supplement is not suitable for clinical use.

Grant Funding Source: Supported by Saving Lives at Birth
Food insecurity is related to obesity and lipid alterations in Mexican college students (805.2)
Authors: Anaya‐Loyola Miriam , González‐Martínez Miguel , Aguilar‐Galarza Beatriz , Moreno‐Celis Ulisses , Sánchez‐Ugalde Diana , Melgar‐Quiñonez Hugo
Published: June 2021

ELCSA (Encuesta Latinoamericana y del Caribe sobre Seguridad Alimentaria) has been validated and used in México for estimating Food Insecurity (FI). The objective of this study was to assess food insecurity in students from the Autonomous University of Queretaro, in México. A total of 662 students answered the ELCSA questionnaire, a food frequency questionnaire in addition, to a nutritional assessment based on weight, height, waist circumference, body composition and biochemical indexes including anemia, glucose, triglycerides and cholesterol. The 50.5% of students had some degree of FI, mainly emphasizing the mild (39.4 %). A higher prevalence of overweight and obesity were observed in students reporting severe‐FI than in those with mild and moderate FI. In contrast , the percentage of students with high body fat decreased with increasing severity of FI , even if the average value of % BF was not statistically different between IF‐students with mild and moderate (24.9 ±7.5 vs. 25.0±7.5, P > 0.05). Anemia affected 1.1% of students, being more prevalent the micro and macrocytic types in those students with FI. Severe FI‐students had the greatest number of cases with hypertriglyceridemia (33.3 %) and hypercholesterolemia (16.7%). According to the findings in this study, FI in Mexican college students could be an important factor related to the occurrence of overweight and obesity and its associated comorbidities among which include hypertriglyceridemia and hypercholesterolemia.

Grant Funding Source: Supported by the Autonomus University of Queretaro