bluuebunny/crossref_metadata_2025 huggingface dataset:
Are Monthly Food Insecurity Assessments Necessary? A Longitudinal Analysis in Rural Zambia
Authors: Na Muzi , Caswell Bess L. , Talegawkar Sameera A. , Palmer Amanda
Published: November 2023

In resource‐limited settings, we generally assume that household food insecurity (HFI) status is static over time, ranging from a period of a few months to a few years. Longitudinal HFI measures are required to test the validity of this assumption. We assessed HFI on a monthly basis in the context of an efficacy trial of biofortified maize conducted in rural Zambia from October 2012 to March 2013. This period is mostly characterized as the lean season, which peaks in ~April in this setting. The trial included two intervention arms and a parallel non‐intervened group for the purposes of background dietary and food security measures. A total of 907 households were enrolled at baseline; however, the present analysis includes longitudinal data from 157 households in the non‐intervened arm. At each of the six monthly visits, HFI was assessed using a validated 8‐item perception‐based Likert scale with a one month recall period. We calculated a HFI index by summing scores over the Likert scale (ranging from 0 to 32). The median (IQR) HFI index was 7 (4–11), 8 (5–11), 7 (4–10), 6 (4–9), 6 (4–9) and 7 (4–9) at the six monthly visits, respectively. The correlation between HFI index between visits ranged from 0.3 to 0.4. A random effect model was fit on the index as a function of visit clustered at household level. Results from the model showed that the HFI index decreased on average by 0.25 points per visit. Within and between household variation in the index was 10.6 and 8.8, respectively. Within‐household variability of the HFI index did not show a clear pattern in relation to the household mean score. Because our repeated HFI measurements were conducted in one single season, the small change in mean monthly HFI index suggested that seasonal assessments may be sufficient for population monitoring purposes in similar settings. However, higher variation within some households suggests that household characteristics—as opposed to seasonal impacts on food security—may explain the differences in variation in response to the repeated scale.

Support or Funding Information

HarvestPlus (grant 8251) and Sight and Life Global Nutrition Research Institute at Johns Hopkins University
A Folate, Vitamin B12 and Homocysteine‐related dietary pattern and risk of subclinical atherosclerosis: findings from the MRC National Survey of Health and Development (NSHD)
Authors: Maddock Jane , Ambrosini Gina L , Ray Sumantra ,   NSHD Study Team
Published: November 2023

Elevated homocysteine concentrations have been shown to increase the risk of stroke. However, the relationship between dietary patterns (DP) associated with homocysteine and associated nutrients (i.e. folate and vitamin B12) with subclinical atherosclerosis, an early marker of stroke, remains unknown. This study identifies a DP characterised by plasma homocysteine, dietary folate and vitamin B12, and investigates its association with carotid intima‐media thickness (cIMT).

Food consumption in the NSHD, a British birth cohort, was assessed using 5‐day estimated food diaries at 36 years (y), 43y, 53y and 60–64y. High resolution cIMT in the right and/or left common carotid artery was measured amongst 1565 participants at 60–64 years. Reduced rank regression was used to identify DPs based on plasma homocysteine (μmol/L), dietary folate (ug/kcal) and vitamin B12 (ug/kcal) as response variables. Multiple linear regression models examined cross‐sectional and prospective associations between DP z‐scores.

The DP explaining the maximum amount of total variation (4.5%) in all response variables was correlated with high intakes of folate (r=0.96) and vitamin B12 (r=0.27) and weakly correlated with homocysteine (r =0.10). The DP was characterised by high intakes of vegetables, fruit and breakfast cereal and low intakes of processed meat, white bread, sugar and preserves. No cross‐sectional association was observed between DP z‐scores and left (0.34% 95% CI −1.09%, 1.79%) or right cIMT (0.07%, 95%CI −1.28% to 1.45%), adjusting for age, gender, dietary misreporting, socioeconomic status, BMI, smoking, physical activity, lipid‐lowering/antihypertensive medication and diabetes. Similarly, there was no association between DP z‐scores measured at the ages of 36y, 43y, 53y and cIMT at 60–63y.

No cross‐sectional or prospective associations were observed between cIMT and a DP high in dietary folate and vitamin B12 in this study. However, the observed DP explained only a small amount of variation in response variables and food intake. Further examination of DPs explaining greater amounts of variation in alternative response variables and their relationships with cIMT is warranted.

Support or Funding Information

Medical Research and British Medical Association, Josephine Landsell Award
The Early Life Predictors of a Dietary Pattern Linked With Greater Metabolic Risk in Adolescence and Tracking of Its Food Components
Authors: Appannah Geeta , Pot Gerda Karolien , Oddy Wendy Hazel , Jebb Susan Ann , Ambrosini Gina Leslie
Published: November 2023

Background

Growing evidence suggests that dietary behaviours linked with chronic diseases in adulthood may develop early in life. However, little is known on the tracking of food intakes during adolescence. This study examined predictors and tracking of key food groups in a dietary pattern (DP) associated with greater cardiometabolic risk among adolescents aged between 14 and 17 y in the Western Australian Pregnancy (Raine) Cohort study.

Methods

A total of 860 adolescents reported their food consumption using a food frequency questionnaire (FFQ) at 14 and 17 y. Z‐scores for an ‘energy dense, high fat and low fibre’ DP were estimated using reduced rank regression (RRR) at both ages. Information on adolescents’ early life exposures and parental socio‐economic status at 14 and 17 y was collected using a detailed questionnaire. Tracking analysis was performed based on the predictive value (PV) of remaining in the same quartile of the DP z‐score between 14 and 17 y. Associations between the DP z‐scores, early life factors and SES were analysed using multiple linear regression.

Results

The identified DP was prospectively associated with greater cardiometabolic risk factors, in boys and girls, between 14 and 17 y. Tracking was strongest amongst boys with the lowest consumption of vegetables, fruit, legumes, high‐fibre bread and yoghurt at 14 y. A high DP z‐score and high intake of processed meat, low‐fibre bread, crisps and savoury snack also tracked strongly among boys. Greater maternal education (β=0.09, p=0.002 at 14 y; β=0.14, p<0.001 at 17 y) and age at child birth (β=−0.09, p=0.003 at 14 y; β= −0.11, p=0.004 at 17 y) were consistently associated with a higher DP z‐score. Paternal smoking status during pregnancy was associated with a higher DP z‐score (β=0.10, p=0.049), at 14 y.

Conclusion

These findings highlight subgroups of the population who may benefit the most from early interventions to improve dietary behaviors and reduce the risk of later chronic diseases.

Support or Funding Information

Medical Research Council, UK (U105960389)

National Health and Medical Research Council of Australia

National Heart Foundation of Australia

Beyond Blue Cardiovascular Disease of Australia

Depression Strategic Research Program of Australia
The Role of miR‐378*, miR‐218 & miR‐351 in Cardiac Hypertrophy
Authors: Sadiq Shahzad , Charchar Fadi J , Crowley Tamsyn , Delbridge Lea , Harrap Stephen , Lewandowski Paul
Published: November 2023

Introduction

Ventricular hypertrophy is an adaptation that is necessary as part of natural growth. It may also occur in response to increased resistance to blood flow (pressure‐overload) as seen in hypertension. The Hypertrophic Heart Rat (HHR) is a model of pressure‐overload cardiac hypertrophy in the absence of hypertension and is thus suitable to study genetic components of pathological increase in cell size without progression to heart failure. Several genes are implicated in the cardiac hypertrophic pathway, which are regulated by microRNAs. The regulatory potential of microRNAs can be exploited to manipulate the expression of target genes. Moreover, microRNAs can also serve as diagnostic/prognostic biomarkers. The study aims to determine microRNAs and their role in cardiac hypertrophy using the HHR.

METHODS

Heart samples were carefully harvested from rats. A preliminary microRNA microarray analysis was performed. RNA was extracted from the heart samples was reverse transcribed, the cDNA quantified and finally expression levels of microRNAs and cardiac biomarkers measured using qPCR.

RESULTS

Expression of miR‐378*, miR‐218 & miR‐351 was decreased in adult HHRs compared to the control. Interestingly, transcription of miR‐378* & miR‐351 was higher in 2‐day old HHRs. Moreover, expression of miR‐218 & miR‐351 were higher in 2‐days old HHRs compared to the adults. Significant differences in expression of cardiac biomarkers of hypertrophy were also noted. Further validation of these microRNAs as regulators of cardiac hypertrophy is currently being examined using a myoblast cell line, H9c2(2‐1), treated with Angiotensin II, which is a recognized stimulant for hypertrophy. These cells are being transfected to up and down regulate the expression of the microRNAs under investigation.

CONCLUSION

Variable expression of the microRNAs at two important time‐points in life reveals their importance as potential targets for anti‐sense therapy and as biomarkers of cardiac hypertrophy. Apoptosis during fetal life (2 day old) is believed to result in fewer cardiomyocytes at birth with consequential compensatory remodeling during adulthood. Further studies are being done on various cardiac cell lines.

Support or Funding Information

School of Medicine, Deakin University, Waurn Ponds, Australia
Fenofibrate Induces Cardiac Fibrosis in Mice Lacking the Co‐chaperone and E3‐ubiquitin ligase CHIP
Authors: Ravi Saranya , Willis Monte S , Lockyer Pamela , Patterson Cam , Schisler Jonathan C
Published: November 2023

Objective

To determine if stimulation of PPARα with fenofibrate can rescue cardiac hypertrophy induced by energy starvation in CHIP−/− mice.

Carboxyl Terminus of HSP70 (CHIP) is a co‐chaperone and E3 ubiquitin ligase involved in protein quality control. The genetic depletion of CHIP in mice is associated with cardiac hypertrophy and decreased survival following myocardial infarction. We previously demonstrated that pressure overload‐induced cardiac hypertrophy is a result of energy starvation and lack of AMPK activation. We found that while the rate of fatty acid uptake in isolated adult mouse cardiomyocytes lacking CHIP expression (CHIP−/−) was similar to WT cardiomyocytes, there was a 33% decrease in fatty acid oxidation accompanied by a 98% increase in fatty acid incorporation into cellular lipids in CHIP−/− versus wild‐type cardiomyocytes. This suggested that a decrease in fatty acid oxidation may contribute to the cardiac disease seen in CHIP−/− mice, and activation of fatty acid oxidation may ameliorate the cardiac pathophysiology associated with the loss of CHIP function. To test our hypothesis, we treated mice with 65 mg/g body weight of fenofibrate, a PPARα agonist that increases lipid uptake and oxidation, for six weeks and observed changes in cardiac function via conscious cardiac echocardiography and histological characterization. Much to our surprise, we found that fenofibrate increased cardiac fibrosis in the CHIP−/− mice by 60%, without a change in total heart size or cardiomyocyte cross‐sectional area compared to wild‐type mice. We determined the effectiveness of fenofibrate as a PPARα agonist by measuring PPARα target genes in the liver of mice and observed similar activation of fatty acid gene expression in both CHIP−/− and wild‐type mice. Likewise, candidate PPAR‐responsive genes in whole heart did not reveal any genotype‐specific differences. We suspect that the fibrotic response is due to pathophysiological stimulation of cardiac fibroblasts lacking CHIP expression. We are now studying isolated primary cardiac fibroblasts from wild‐type and CHIP−/− mice in combination with PPAR isoform specific agonists to measure the fibrotic response to PPAR activation. Our findings have important implications for understanding how the loss of CHIP function (CHIPopathy) recently identified as the causal gene in spinocerebellar ataxia autosomal recessive 16, affects cardiac function given the large population of patients taking fibrates to treat hyperlipidemia.
Loss of CHIP Expression Perturbs Glucose Homeostasis and Leads to Type II Diabetes Through Defects in Microtubule Polymerization and Glucose Transporter Localization
Authors: Lenhart Kaitlin Christine , McDonough Holly , Ronnebaum Sarah M. , An Jie , Newgard Christopher B. , Willis Monte S. , Patterson Cam , Schisler Jonathan Cummings
Published: November 2023

Glucose homeostasis requires insulin signaling throughout the body to maintain physiological blood glucose levels. The loss of insulin sensitivity, or insulin resistance, is associated with a myriad of diseases including Type II diabetes, atherosclerosis, abdominal obesity, and hypertension. Cellular protein quality control pathways involving the chaperone and ubiquitin proteasome system have been associated with the pathophysiology of insulin resistance, but the molecular details are incompletely understood. We and others have previously demonstrated a role for the co‐chaperone and E3‐ubiquitin ligase CHIP (Carboxyl terminus of HSC70‐Interacting Protein) in the regulation of several metabolism‐associated proteins including AMPK, SirT6, AKT, SGK‐1 and PTEN. We demonstrate that mice genetically deficient in CHIP (gene locus Stub1) exhibit a striking Type II diabetes‐like phenotype, including impaired glucose tolerance, decreased insulin sensitivity, and decreased insulin‐stimulated glucose uptake in skeletal muscle in comparison to wild‐type animals. Interestingly, CHIP‐depleted C2C12 skeletal myoblasts exhibited a profound defect in the translocation of the glucose transporter Glut4 to the plasma membrane after glucose/insulin stimulation. The impairment of Glut4 translocation was accompanied by a decrease in both tubulin polymerization and phosphorylation of stathmin (Ser16), a microtubule‐regulating protein required for polymerization‐dependent protein trafficking. PIP3, required for microtubule reorganization, was sustained in wild‐type C2C12 cells following stimulation whereas CHIP‐depleted C2C12s exhibited only transient increases in PIP3 that diminished 30 min after stimulation. Moreover, CHIP‐depleted C2C12s exhibited a concomitant reduction in the levels of phosphorylated AKT (Thr308), a signaling event that requires PIP3‐dependent translocation of AKT to the plasma membrane. The sustained PIP3 levels in wildtype C2C12s correlated with decreasing PTEN levels, a negative regulator of PIP3, at a time when CHIP and PTEN co‐immunoprecipitate. Conversely, a decrease in PTEN is not observed in CHIP‐depleted cells. Together, these data describe a novel role for CHIP in an insulin induced phosphorylation signaling cascade regulating microtubule polymerization necessary for glucose transporter translocation, thereby promoting glucose homeostasis and sensitivity to insulin.
Metabolomic Profile of Serum Retinol Concentrations in the ATBC Study Cohort
Authors: Albanes Demetrius , Panagiotou Orestis , Weinstein Stephanie , Moore Steven , Mondul Alison
Published: November 2023

The Alpha‐Tocopherol, Beta‐Carotene Cancer Prevention (ATBC) Study of male smokers found that beta‐carotene supplementation (20 mg/day for 5–8 years) led to an 18% increase in lung cancer incidence, as well as a 5% increase in serum retinol. Higher serum retinol concentrations have also been associated with elevated prostate cancer risk in this and other prospective cohort analyses (AM Mondul et al. Am J Epidemiol 2011;173:813–21, and TJ Key et al. Am J Clin Nutr 2015;102:1142–57). In order to provide biochemical leads regarding how vitamin A status might influence the risk of developing this malignancy, we conducted an agnostic analysis to examine the metabolomic profile of circulating retinol concentrations. The study is based on pre‐randomization fasting serum from 1,282 ATBC Study participants, retinol determined by high‐performance liquid chromatography, and metabolites identified using ultrahigh‐performance liquid chromatography/mass spectroscopy and gas chromatography/mass spectroscopy (Metabolon, Inc., Durham, N.C.). Raw data were extracted, peak‐identified, and quality control processed, and 800 identified compounds were analyzed, with a median metabolite coefficient of variation of 9%. Linear multivariate regression models computed the standardized beta‐coefficient and 95% confidence intervals (CI) of the log‐transformed metabolite‐retinol associations. Using p=0.05/800 (6×10−5) as the significance threshold to adjust for multiple comparisons, 65 metabolites were significantly associated with fasting serum retinol (units: micrograms/liter). The strongest associations (p<10−10), all positive, were for N‐acetyltryptophan (β=0.001), 1‐palmitoleoylglycerophosphocholine (β=0.001), 1‐palmitoylglycerophosphoethanolamine (β=0.0008), 4‐acetamidobutanoate (β=0.0006), myoinositol (β=0.0004), 6‐carbamoylthreonyladenosine (β=0.0004), and urate (β=0.0003). Androgen metabolites were also positively associated with retinol concentration (p=10−9); for example, 5alpha‐androstan‐3beta,17beta‐diol sulfate (β=0.002) and 4‐androsten‐3beta,17beta‐diol monosulfate (β=0.002). How these and other phospholipid, amino acid, and androgen metabolites are related to retinol metabolism, and whether any of them directly influence prostate cancer risk, will require further investigation. Similar metabolomic profiling studies of serum retinol in other populations would be useful.

Support or Funding Information

Research support: Intramural Research Program of the National Cancer Institute, NIH
Parietal Valve of the Coronary Sinus: Morphological Features and Clinical Importance
Authors: de Prates Nadir Eunice Valverde Barbato
Published: November 2023

Retrograde cardioplegia is a procedure regularly used to protect the miocardium during cardiac surgeries. The cardioplegic solution is retrogradly perfused through the coronary sinus, the main vein of the heart, and it is useful since the coronary venous system has shown to be free of any obstructing atherosclerotic disease. The opening of the coronary sinus occurs directly in the right atrium and an ostial valve at this level has long been related. However, a parietal valve in the coronary sinus has renewed the interest in this study. These valves could make difficult a perfect perfusion of the cardioplegic solution, particularly when low coronary sinus pressures are used. In order to understand the morphology of the parietal valve of the coronary sinus we decided to perform a study of the valve under SEM. Six fragments of the coronary sinus containing the parietal valve were taken from adult individuals, post mortem, with no gross cardiac pathologies. The samples were submitted to the usual techniques of Scanning Electron Microscopy. The parietal valve of the coronary sinus showed a half‐moon shape, with a well defined structure. Its endothelium was arranged in parallel lines, similar to a stem of a tree, where we could evidence the nuclei of the endothelial cells. Under the endothelial layer we found abundant collagen and elastic fibers. Owing to its developed form seen under SEM, the parietal valve of the coronary sinus must play an important role in guiding the blood flow through the coronary sinus to the right atrium in physiological conditions and could raise difficulties to the flow of the cardioplegic solution during the retrograde cardioplegia.
Reelin Signaling in Cardiovascular Development
Authors: Kim Dongwook , Poles Asha , Gorski Grzegorz , Hatcher Cathy J
Published: November 2023

The epicardium and coronary vessels are derived from a cell lineage that descends from the primitive proepicardium during cardiovascular development. These structures are formed through developmentally linked processes that are regulated by a network of transcription factors that control cell behavior and molecular genetic expression. Previously, we demonstrated that the Tbx5 transcription factor contributes to cell adhesion during epicardium formation and coronary vasculogenesis. We speculate that this occurs through potential transcriptional regulation of the Reln gene by Tbx5. Reelin, which is encoded by the Reln gene, is a secreted, extracellular matrix glycoprotein that has an established role in neural and lymphatic system development where it regulates cell migration and positioning. However, its role in cardiovascular development has yet to be determined. Our goal is to identify the functional contributions of reelin signaling to the structural formation of the developing heart and embryonic cardiac cell behavior. We used immunofluorescent staining to examine the developing mouse heart for expression of reelin as well as Crk, an adaptor protein which is a downstream mediator of the reelin signaling pathway. Reelin expression is detected in the proepicardium, myocardium and endocardium of the developing mouse heart as early as embryonic day (e) 9.5. At later developmental stages, reelin is expressed in the epicardium as well as specific cell types within the heart, including endothelial cells. Crk expression is detectable as early as e11.5 in the epicardium and myocardium. These findings prompted us to explore the physiological role of reelin at the cellular level. We assessed its contributions to in vitro cell behavior in the human microvascular endothelial cell (HMEC) line that models aspects of coronary vasculogenesis. Primary cultured HMECs were treated with RELN‐specific small interfering RNAs to knock down endogenous reelin expression. Preliminary results indicated that reelin knockdown caused detectable changes in cell behavior. Together, the results of these in vitro and in vivo studies provide insight into how reelin signaling contributes to cellular functions and mechanisms required for cardiovascular development.

Support or Funding Information

PCOM Center for Chronic Disorders of Aging
Imaging cell‐cell interactions with bioluminescent probes
Authors: Prescher Jennifer
Published: November 2023

Cell‐cell contacts and interactions drive numerous physiological processes, including immune function, neurotransmission, and cell migration. Breakdowns in cell‐to‐cell communication also underlie numerous pathologies. While cellular interactions play key roles in human health and disease, the mechanisms by which cells transact information in vivo are not completely understood. The number of cells types involved, the timing and location of their interactions, the molecular cues exchanged, and the long‐term fates of the cells remain poorly characterized in most cases. This is due, in part, to a lack of tools for observing collections of cells in their native habitats. My group is developing novel bioluminescent probes to “spy” on cells and decipher their communications in vivo. In one area, we are crafting bioluminescent probes for multi‐cellular imaging in whole organisms. In a second realm, we are generating tools that selectively illuminate cell‐cell contacts. These imaging strategies will be discussed, along with their application in mapping immune cell interactions with tumor tissue and disseminated pathogens.
Regulation Of Lipid Metabolism By SREBPs
Authors: Espenshade Peter
Published: November 2023

Lipid synthesis requires molecular oxygen. Consequently, maintenance of cell growth under hypoxic conditions requires adaptive mechanisms that control lipid homeostasis. In mammalian cells, sterol regulatory element‐binding protein (SREBP) transcription factors are central regulators of cellular lipid homeostasis. Our studies in fungi demonstrate that SREBPs are oxygen‐responsive transcription factors required for adaptation to low oxygen and for host infection by pathogenic fungi. In fission yeast, proteolytic release of SREBP in response to low oxygen requires the Golgi membrane‐anchored Dsc E3 ligase complex, but no SREBP protease has yet been identified. Recently, we identified an uncharacterized protease required for SREBP proteolytic processing, and we will present our results on how this protease functions in SREBP activation. In addition, fission yeast SREBP does not activate all lipid synthesis genes under low oxygen, for example, those required for unsaturated fatty acid synthesis. We will present data demonstrating that the transcription factor Mga2 is also oxygen‐responsive and functions as an SREBP‐like transcription factor during hypoxic adaptation. Collectively, these two pathways control lipid homeostasis in response to low oxygen in fission yeast.

Support or Funding Information

Studies were supported by the National Institutes of Health (HL‐077588).
Utilizing Smartphone Technology To Monitor Improvements In The Healthiness Of The Food Supply
Authors: Dunford Elizabeth , Crino Michelle , Neal Bruce
Published: November 2023

Purpose

To date, there has been little success in efforts to stem the tide of diet‐related ill health attributable to increased availability of highly processed foods providing high levels of saturated fat, sugar and salt. Central to the challenge faced by policy makers in addressing this is the absence of current data to properly define, understand or respond to the problem.

Methods

The Food Monitoring Group was established in 2010 with the objective of collecting data describing the nutritional composition of the global packaged food supply. The Group harnessed smartphone technology to develop a database that contains brand‐specific information on the nutritional composition of >250,000 foods globally. The “Data Collector App” is made available to all countries around the world to enable simple, low‐cost collection of food composition data on the packaged food supply.

Results

Data have been used to monitor the global food supply. E.g. data from Australia were used to monitor the implementation of government sodium reduction targets. Comparisons of sodium levels over time exposed the limited progress in achieving the targets, with data presented at the individual company level. Data from India highlighted the incompleteness of nutritional labelling as another key issue in the field and was used to push government to better enforce existing labelling standards in the country. Data are also being used to fuel the FoodSwitch smartphone application, which allows consumers in Australia, NZ, the UK, China, India and South Africa to scan the barcodes of food items in‐store and be directed to healthier brands of similar products. If a product is missing from the database, app users are prompted to take 3 photos of the missing item. This alone has resulted in more than 700,000 photos being sent in by users to date, and a minimum of 300 new photos being submitted daily. These crowd‐sourced data are also used to help drive improvements in national food supplies. The app already has more than 760,000 downloads in Australia, NZ and the UK alone, with launches in the USA, Canada, Hong Kong and Switzerland planned for 2016.

Conclusion

Smartphone technology has helped collect large amounts of information about the healthiness of the global food supply. These data have been used for both monitoring of the nutritional content of foods, government initiatives to improve the food supply, and existing labelling initiatives. The data have also been used to educate consumers through the FoodSwitch smartphone application, which in turn helps crowd‐source additional data on the food supply, thus allowing for low‐cost, real‐time tracking of the healthiness of the global food supply.

Support or Funding Information

E Dunford is supported by a NHMRC Early Career Fellowship.
Correlation of Adolescent Caffeine Intake with Blood Pressure in Adolescence and Adulthood
Authors: Estrada Erika , Urbina Elaine M , Daniels Stephen R , Woo Jessica G
Published: November 2023

Objective

High blood pressure (HBP) is one of the leading causes of premature death worldwide. Currently, 1 in 3 adults has HBP. Recent studies have shown that caffeine leads to an acute increase in blood pressure after exposure, but the long–term effects of caffeine on blood pressure are unclear. Our study aimed to assess the longer‐term relationships between dietary caffeine intake and blood pressure using data collected in two longitudinal epidemiologic studies: the Princeton Lipid Research Study and the NHLBI Growth Health Study (NGHS).

Methods

The Princeton cohort included 623 subjects who were seen as children/adolescents and again in adulthood. The NGHS cohort included 870 black and white females ages 9 or 10 at enrollment who were followed yearly for up to ten years. Caffeine intake was assessed using a 24‐hour recall for Princeton (childhood only) and 3‐day diet records for NGHS (at 8 of 10 visits). Blood pressure in both studies was assessed using standard mercury sphygmomanometer, and systolic and diastolic z‐scores were calculated for childhood measurements. The relationship between caffeine intake and blood pressure was analyzed for individuals in each visit.

Results

In the NGHS cohort, caffeine intake increased from a median of 15 mg to 40 mg/day between ages 10 and 19 years, and was significantly higher in white vs. black participants at all ages (p<0.0001). Similar patterns were noted in Princeton. In NGHS, SBP z‐score and DBP z‐scores were higher in black vs. white participants from age 17–19 (p<0.01); in Princeton, BP z‐scores did not differ by race at any childhood age. Cross‐sectionally, neither cohort demonstrated significant correlations between caffeine intake and blood pressure z‐scores during childhood/adolescence, except for a negative correlation seen in NGHS with DBP z‐score at age 16.0 (Spearman correlation=−0.085, p=0.04). Longitudinally, in Princeton, caffeine intake in childhood/adolescence was not associated with systolic or diastolic blood pressure in adult follow up (both p>0.4), adjusting for sex, race, and age and BMI at adult follow up. In NGHS, neither caffeine intake at age 12, nor the change in caffeine intake between ages 12 and 19, were associated with blood pressure z‐scores at participants’ final visits, adjusting for age at final visit and BMI z‐score.

Discussion

The results of these two long‐term studies suggest that caffeine consumption during childhood/adolescence is not associated with blood pressure in either later adolescence or adulthood.

Support or Funding Information

EE supported by a grant from the American Heart Association and the CCHMC Schmidlapp Young Women's Scholarship.
Mango (Mangifera Indica L.) in the promotion of intestinal regularity and decreases inflammation in human subjects with constipation
Authors: Mertens‐Talcott Susanne U , Kim Hyemee , Talcott Stephen T , Venancio Vinicius Paula
Published: November 2023

Chronic constipation is a common gastrointestinal condition associated with intestinal inflammation and a considerably impaired quality of life, affecting approximately 10–20% of the population in the US. Dietary fiber and laxatives have been shown to aid in the treatment of chronic constipation. In this study, the consumption of mango fruit was compared to an equivalent amount of fiber in its effect on abdominal distention and constipation in otherwise healthy human volunteers. The participants were randomly divided into either the experimental mango group or control fiber group. The mango group consumed 300 g of mango daily for 4 weeks, while the fiber group consumed an equivalent fiber amount (1 teaspoon). Questionnaires regarding intestinal evacuation and digestive wellness were collected weekly, and blood and stool samples were collected at the beginning of the study and after four weeks. The results show that the mango group had better improvement of constipation status as compared to the fiber group, based on evacuation categorization (stool frequency, consistency, and shape). Additionally, increase in gastrin and Peptide YY (PYY) were higher in the mango group than the control fiber group. The production of endotoxin and IL‐6 were lower in subjects who received mango treatment. In response to the mango intake, two fecal short chain fatty acids, acetic and valeric acid, were increased. Compared to the consumption of fiber, the consumption of mango in the treatment of chronic constipation had higher subject adherence, improved parameters of intestinal evacuation, reduced the production of endotoxins, reduced inflammation, and increased the concentration of short chain fatty acids, all of which have been established to contribute to intestinal health and wellness.

Support or Funding Information

Mango National Board
Determination of the in vivo and in vitro protein quality of pulse protein concentrates and isolates.
Authors: Nosworthy Matthew G , Neufeld Jason , House James D
Published: November 2023

Currently, the use of a rodent bioassay is the officially recognized method for generating data in support of protein content claims for food ingredients. Although there are in vitro methods for calculating protein digestibility, to date no method has been accepted by North American regulatory agencies for use in establishing protein content claims, likely due to a lack of evidence in support of their validity. To this end, we compared the in vivo (%True Fecal Protein Digestibility; %TFPD) and the in vitro protein digestibility (%IVPD) of faba bean, lentil and pea protein concentrates and isolates. Sprague‐Dawley rats (n=70, ~70 g) were randomized to one of seven diets (10% crude protein) based on: casein (reference control), the concentrates of faba bean (FB60), lentil (L55) or pea (P55) or the protein isolates of the same pulses (FB85, L85 and P85 respectively). The %TFPD and protein digestibility corrected amino acid score (PDCAAS) were calculated. The %IVPD was determined via enzymatic digestion with subsequent pH drop, and used to calculate the in vitro PDCAAS (IVPDCAAS). Except for L55, in which the sulfur amino acids were limiting, all experimental diets were limited by tryptophan content. FB60 had a higher AAS than FB85 (0.46 vs 0.58) whereas the AAS of the other diets had the opposite trend (L55‐0.75 vs L85‐ 0.52, P55 – 0.58 vs P85‐ 0.54). Digestibility was greater in the protein isolates than the concentrates, and the in vivo rat bioassay reported higher digestibility than the in vitro method; casein (95 vs 84), FB60 (94 vs 74), FB85 (97 vs 75), L55 (91 vs 75), L85 (96 vs 77), P55 (93 vs 78), P85 (97 vs 78). There was no correlation found between %TPD and %IVPD (R2= 0.027). Interestingly, a strong correlation was found between PDCAAS and IVPDCAAS (R2=0.99, p<0.0001), however the values from the rodent model were greater than the in vitro casein (89 vs 79), FB60 (43 vs 33), FB85 (56 vs 44), L55 (68 vs 56), L85 (50 vs 40), P55 (54 vs 46), P85 (53 vs 43). Excepting faba bean, all pulse concentrates had higher AAS, lower digestibility and greater PDCAAS values than their isolate counterparts. As such, processes used in the isolation of pulse protein sources increased digestibility, but may have led to shifts in protein composition, leading to a lower PDCAAS value. Although the strong correlation found between PDCAAS and IVPDCAAS suggests that in vitro analysis could reduce the requirement of animal experimentation for protein claims, a larger dataset combined with investigation into other potential in vitro methods is required.

Support or Funding Information

Pulse Canada
Evaluation Of Dietary Assessment Tools: Does ‘Validated’ Mean What We Think It Means?
Authors: Kirkpatrick Sharon I , Subar Amy F , Krebs‐Smith Susan M. , Thompson Frances E , Reedy Jill , Schap TusaRebecca E , Vanderlee Lana , Robson Paula , Csizmadi Ilona , Boucher Beatrice A , Massarelli Isabelle , Rondeau Isabelle
Published: November 2023

Evaluation of the extent to which self‐report dietary assessment tools accurately capture true intake is critical to informing the collection of dietary intake data that can advance our understanding of diet and health. Such evaluation is challenging due to the lack of unbiased reference measures for dietary intake, forcing reliance on self‐report (and thus error‐prone) measures as comparators. Depending on the type of instrument and its intended use, evaluation is also complicated by the need to test tools in different settings and populations. The objective of this work is to examine challenges in evaluating dietary assessment tools and the ways in which the findings of validation studies are interpreted. We draw upon a scoping review of Canadian studies of free‐living adult populations that included an assessment of dietary intake to illustrate the extent and ways in which validity of tools are addressed. Of 58 studies that used a frequency questionnaire or screener to assess diet, tools were reported to have been validated for the given study sample in a small subgroup. In close to two‐thirds, authors reported that the tool has been validated, in some cases with few details on how it was evaluated and in what population(s). In close to a third, there was little attention to whether tools used have been tested. Given that self‐report instruments cannot be expected to perfectly capture true intake, more nuanced discussions of what it means for a tool to be ‘valid’ as well as the implications of limitations in tools for study results appear warranted. Such discussions are key to improving the quality of self‐reported dietary intake data.

Support or Funding Information

This scoping review was supported by the Canadian Institutes of Health Research (137238) and Canadian Cancer Society Research Institute (702855).
Disease Associated Polymorphisms in MnSOD and GPx‐1 Affect Metabolism, Mitochondrial Membrane Potential and Expression of Signaling Proteins
Authors: Ekoue Dede N , Bera Soumen , Ansong Emmanuel , Hart Peter , Macias Virgilia , Kajdacsy‐Balla Andre , Bonini Marcelo , Diamond Alan M
Published: November 2023

MnSOD detoxifies superoxide and impacts tumor biology by generating H2O2 which can diffuse through the mitochondrial membrane and affect metabolism and apoptosis. The selenium‐dependent enzyme GPx‐1 localizes to both the cytoplasm and the mitochondria and reduces H2O2 to water and may therefore modulate the impact of MnSOD on carcinogenesis. A val to ala polymorphism in codon 16 of the MnSOD gene has been shown to be associated with increased prostate cancer risk in men with the lowest level of dietary antioxidant intake and individuals who consumed less dietary antioxidants, including selenium, had the greatest risk of prostate cancer. A polymorphism in the GPx‐1 gene resulting in a leucine (L) instead of a proline (P) at position 198 has been reported to be associated with elevated cancer risk. To examine the molecular mechanism behind the epidemiological observation that genotypes in GPx‐1 gene modify elevated risk of cancer associated with MnSOD genotypes, MCF‐7 human breast cancer cells null for GPx‐1 and having negligible levels of endogenous MnSOD were transfected with GPx‐1 and MnSOD allele‐specific expression constructs to determine outcomes related to cellular signaling and metabolism. MnSOD and GPx‐1 alleles differentially interacted to modulate the expression of the anti‐oxidant stress response regulator Nrf2, the cell adhesion protein E‐Cadherin, the cell signaling protein pAkt, the anti‐apoptotic protein Bcl‐2 and the mitochondria located deacetylase Sirt3. Also, co‐expression of the GPx‐1A7L and either MnSODval or MnSODala increased oxidative phosphorylation as measured by O2 uptake using the Seahorse XF24 XF analyzer, which simultaneously determines relative mitochondrial respiration and glycolysis. In order to assess whether GPx‐1 and MnSOD allelic variants modulate mitochondrial membrane potential, CMX‐ROS fluorescence was measured. Independent and irrespective of which allele was evaluated, MnSOD and GPx‐1 individually decreased mitochondrial potential as compared to that seen using MCF‐7 control cells. In addition, co‐expression of GPx‐1A7L with either MnSODval or MnSODala further decreased membrane potential above that observed for either MnSOD and GPx‐1 alone. In order to determine if MnSOD and GPx‐1 levels are associated with a higher risk for biochemical recurrence of prostate cancer after radical prostatectomy, immunohistochemistry of human prostate tissue cores will be performed. Preliminary results indicate that a high MnSOD/GPx‐1 ratio was observed in prostate cancer tissue compared to adjacent normal tissue.

Support or Funding Information

This research was supported by the National Cancer Institute of the National Institutes of Health under Award Number R21CA182103 awarded to AMD and DNE.
Overexpression of Ghrelin Is Associated with Poor Prognosis of Renal Cell Carcinoma and Promotes Metastasis through GHSR‐mediated Activation of AKT‐Snail Pathway
Authors: Lin Tsung‐Chieh , Hsu Shih‐Lan , Hsiao Michael
Published: November 2023

Loss of cell‐cell adhesion, increased cell motility, and extracellular matrix (ECM) degradation are critical factors in cancer cell metastasis. In this study, we identified ghrelin as a pro‐metastasis molecule in renal cell carcinoma. Ghrelin is an appetite‐regulating molecule that promotes growth hormone (GH) release and food intake through growth hormone secretagogue receptor (GHS‐R). Ghrelin expression is observed in proximal and distal renal tubules, where renal cell carcinoma (RCC) arises. However, whether ghrelin is up‐regulated and promotes renal cell carcinogenesis remains unclear. In this study, we found ghrelin was highly expressed in renal tumours, especially in metastatic RCC. In addition, high ghrelin levels correlated with poor outcome, lymph node and distant metastasis. Our results also showed the addition of ghrelin promoted the migration ability of RCC cell lines 786–0, ACHN and A‐498. Furthermore, knockdown of ghrelin expression reduced in vitro migration and in vivo metastasis, suggesting a requirement for ghrelin accumulation in the microenvironment for RCC metastasis. Analysis of microarray signatures using Ingenuity Pathway Analysis (IPA) and MetaCore pointed to the potential regulation by ghrelin of Snail, a transcriptional repressor of E‐cadherin. We further observed that Ghrelin increased the expression, nuclear translocation and promoter‐binding activity of Snail. Snail silencing blocked the ghrelin‐mediated effects on E‐cadherin repression and cell migration. Snail–E‐cadherin regulation was mediated by GHS‐R‐triggered Akt phosphorylation at Ser473 and Thr308. Pretreatment with PI3K inhibitors, LY294002 and wortmannin, as well as Akt siRNA, decreased ghrelin‐induced Akt phosphorylation, Snail promoter binding activity and migration. Taken together, our findings indicate that ghrelin can activate Snail function via the GHS‐R–PI3K–Akt axis, which may contribute to RCC metastasis.

(This study was supported by Academia Sinica to Dr. Michael Hsiao.)
Targeting macrophage necroptosis for therapeutic and diagnostic interventions to treat atherosclerosis
Authors: Karunakaran Denuja , Geoffrion Michele , Wei Lihui , Gan Wei , Perisic Ljubica , Maegdefessel Lars , Hedin Ulf , Sad Subash , Virmani Renu , Ruddy Terrence , Rayner Katey
Published: November 2023

Atherosclerosis is a disease of maladaptive inflammation driven primarily by macrophages, and as the atherosclerotic plaque progresses and becomes more complex, a necrotic core forms that ultimately underlies the instability that drives plaque rupture and myocardial infarction. Necroptosis or ‘programmed necrosis’ is an emerging cell death pathway involving kinases RIP3 and MLKL that in contrast to apoptosis, induces a pro‐inflammatory state. We hypothesize that macrophages undergo necroptosis in response to atherogenic stimuli in the vessel wall, and this process underlies necrotic core formation and plaque instability. Here, we show that oxidized LDL (oxLDL) induces necroptotic cell death in macrophages, which can be blocked by Nec‐1, a necroptosis inhibitor. Macrophages treated with oxidized LDL have increased expression of necroptotic genes RIP3 and MLKL through activation of the promoter region and increased RIP3 and MLKL phosphorylation‐ critical steps in the execution of necroptosis. Further, the combined treatment with oxLDL and DAMPs (damage associated molecular patterns) amplified macrophage necroptotic cell death, indicating that additional inflammatory stimuli present in the lesion could act synergistically to promote necroptosis. In a model of established atherosclerosis, Nec‐1 intervention in Apoe−/− mice reduced lesion size (p<0.05) and markers of plaque instability, including reduced necrotic core formation (62.6% reduction, p<0.01). Further, we developed a novel 123I‐Nec‐1 radiotracer targeting the necroptosis pathway and show that it specifically localizes to and tightly correlates with atherosclerotic lesions. Lastly, in humans with unstable carotid atherosclerosis, expression of the necroptotic genes RIP3 and MLKL is elevated and MLKL phosphorylation is detected in advanced atheromas, indicating that the necroptotic pathway is highly activated in vulnerable plaques. In conclusion, our findings offer molecular insight into the mechanisms by which inflammatory atherogenic ligands drives cell death necrotic core formation that underlies unstable atherosclerosis and identify innovative diagnostic and therapeutic tools to treat atherosclerosis.
Ischemia/reperfusion‐induced myocardial injury in mice with diabetes mellitus: role of silent information regulator 1
Authors: Tao Aibin , Mele Tina , Kao Raymond , Martin Claudio , Rui Tao
Published: November 2023

The aims of this study is to determine whether silent information regulator 1 (Sirt1) is involved in the increased susceptibility of the diabetic myocardium to ischemia/reperfusion‐induced (I/R) injury.

Methods

In vivo: mouse model of diabetes was induced by streptozotocin and myocardial levels of Sirt1 assessed (Western). In vitro: isolated cardiomyocytes were conditioned with high glucose (HG) followed by anoxia/reoxygenation (A/R) challenge. Myocyte Sirt1, Dynamin‐related protein 1 (Drp1) expression (Western) and myocyte apoptosis (Caspase 3 activity and cell death ELISA) were assessed.

Results

Myocardial levels of Sirt1 were decreased in mice with diabetes. Sirt1 expression in myocytes with HG was down‐regulated. Myocytes with HG incurred reduced Drp1 S637 phosphorylation and increased apoptosis as compared to naive myocytes challenged with A/R. The effects were prevented by pretreatment of the myocytes with Sirt1 activator or over‐expression of Sirt1 in cardiomyocytes with HG.

Conclusion

Increased vulnerability of diabetic myocardium to I/R injury may be attributed to decreased Sirt1 and subsequent Drp1 activation. (*Corresponding author)

Support or Funding Information

IRF of Lawson Health Research Institute (IRF 2014‐25); National Natural Science Foundation of China (81370333); Natural Science Foundation of Jiangsu Province, China (BK2015‐1332)