bluuebunny/crossref_metadata_2025 huggingface dataset:
Vitamin A Sugar Fortification in Zambia: Opportunities and Challenges
Authors: Greene Matthew , Kabaghe Gladys , Musonda Mofu , Palmer Amanda C
Published: November 2023

Large‐scale food fortification is a major strategy to improve dietary vitamin A (VA) intakes and control VA deficiency. Zambia introduced mandatory sugar fortification with VA, at a target of 10 mg/kg, in 1998; however, there are conflicting reports as to the program's impact on VA status. Our objective was to assess the current state of the program from the policy level to the household. We conducted key informant interviews with stakeholders regarding the program's history, implementation, impact, and perceived challenges. In one district, we mapped formal and informal sugar vendors and recorded coverage of available brands and packaging options. In a random sample, we conducted a Knowledge, Attitudes, and Practices survey of both vendors and consumers. We obtained 50 g samples of sugar from vendors for quantitative VA analysis using an iCheck portable fluorimeter and, in an ongoing surveillance cohort in this community, collected ~1 g samples for qualitative analysis. Key informant interviews revealed a strong commitment to sugar fortification in Zambia, but also several concerns: potential for excess VA intake, poisoning or infertility, cost of fortificant, and limited resources available for monitoring and evaluation of program impact. Market assessments revealed a variety of sources for fortified sugar. The distribution of VA in sugar in market samples was highly skewed, with a median of 2.4 mg/kg (25th percentile, 75th percentile: 1.5, 5.6) and a range from 0.6 mg/kg to 18.0 mg/kg. VA levels in sugar varied by source. Overall, the commitment of stakeholders to VA fortification of sugar is promising for the program's ability to adjust course, as needed, and for its long‐term sustainability. While limited to one area of the country, our data suggest wide variability in VA content, with median levels falling below the 10 mg/kg target. Given observed differences in VA content, it is possible that this is related to varied storage and packaging—a challenge that could be addressed by targeted messages to vendors. Additional research is needed to better understand patterns of sugar consumption at the household level, both to inform the contribution of fortified sugar to overall dietary VA intakes, but also as a baseline in a country that is experiencing a nutrition transition.

Support or Funding Information

Funded by Sight & Life, with support from CBM Internationa
Make‐It‐Yourself (MIY) Food Stations: Long‐Term Healthy Lessons from Nordic Cafeterias
Authors: Tran Huy Quoc , Wansink Brian , Karevold Knut Ivar
Published: November 2023

Do Make‐It‐Yourself (MIY) food stations in the cafeterias lead people to eat healthier or do they lead people to eat more? This three‐month study involving 11,870 lunch guests in nine Nordic hotels shows that 4% more diners selected Caesar salad when it was presented in a “Make It Yourself” station as opposed to premade option. Unfortunately, MIY station also led people to serve themselves 37% more of beef. MIY food stations allow diners to sample more widely, but they also made diners to indulge more in other foods. Although using MIY station can effectively increase the percentage of people who try healthy foods, it may not restrain diners from still over‐serving large amounts of less healthy foods.
Determinants of Dietary Supplement Use and Compliance by US Adults
Authors: Bogale Alemtsehay , MacKay Douglas , Mitmesser Susan H
Published: November 2023

Introduction

Evidence suggests that dietary supplement (DS) use, as a preventive measure to nutrient shortfalls, is among one of the components of a healthy life style.

Objective

To investigate consumer related factors that may affect DS use and compliance, and to recommend consumer centered tools and strategies to improve dietary supplement compliance.

Method

Data from The Council of Responsible Nutrition (CRN) Consumer Survey, published NHANES data and literature review was used to determine the prevalence, gender and age distribution, reasons for DS use and non‐use, and compliance and noncompliance by US adults. These data sources were utilized to compile a comprehensive list of reasons for DS use, reliable information sources of DS, and comparisons of nutrient profiles of DS users versus non‐users.

Results

Overall about 68% Americans use DS. In contrary to earlier findings that suggested increase, DS use remained stable at 68 % for five consecutive years from 2011–2015. Seventy five percent of DS users take multivitamin/mineral supplements followed by Vitamin D (37%), vitamin C (34%), calcium (29%), and vitamin B/B complex (24%). DS use tended to be higher in females (71%) but, compliance was higher in males (69%) than females (60%). Older adults age 55+ had both higher user prevalence (71%) and compliance (75%). Higher DS compliance was observed in the Northeast (68%) and South (69%) than Midwest (59%) and West (58%).

The main reasons of DS use were: overall health (43%), energy (30%), to fill nutrient gap (28%), immune health (23%), bone health (22%), heart health (21%) healthy aging (20%), skin, hair & nails (19%) digestive health (17%) and joint health (17%). Physicians (55%), pharmacists (33%) and nutritionists (30%) were listed as a trusted source of information about DS. The primary reason indicated for noncompliance and non‐use was forgetfulness and not feeling the need. Among DS users, 10% for calcium and 25% for vitamin D did not meet the recommended daily intake, while 38% and 96% non‐users did not meet the daily requirement respectively. Other nutrients that fall short for both DS users and non‐users include potassium, magnesium, zinc, and, vitamins A, C, E and K.

Conclusion

Almost two thirds of Americans use DS for overall health, energy, and to fill nutrient gaps. DS users are more likely to meet the daily recommended nutrient amount than non‐users, however nutrient gaps continue to exist amongst DS users. Non‐compliance appears to be a potential barrier to meeting nutrient recommendations. Improved systems of DS delivery such as simplified regiment with improved sensory attributes and personalized products that target individual nutrient shortfalls may improve DS compliance and contribute to achieving recommended nutrient intakes. Imparting scientific research findings through trusted sources such as health care providers and nutritionists should be considered among the primary strategies in nutrition intervention programs to improve compliance.
Interrelationship between Total and Undercarboxilated Osteocalcin Levels, and Body Mass Index in Women
Authors: Zago Liliana , Infantino Carlos Alfredo González , Zeni Susana , Río María Esther , Felipoff Ana Lía , Weisstaub Adriana , Dupraz Hernán , Presner Natalia
Published: November 2023

Obesity and osteoporosis are two of the main common chronic diseases in women. Both are interconnected and contribute to glucose homeostasis. The objective of this study was to evaluate the effect of BMI on serum levels of total (OC) and undercarboxilated osteocalcin (ucOC), in 100 non diabetic overweight or obese adult women. Women were divided according BMI (range 25.0 to 52.8 kg/m2) into quartiles, and according to their menopausal status: premenopausal (PreM: ≤45 y; n=33), perimenopausal (PeriM: 46–54 y; n=18), postmenopausal (PostM: 55–65 y; n=34) and late menopausal (LM: >65 y; n=15). Total OC, ucOC and leptin were determined by ELISA kits. There were not significant differences between PreM and PostM from the same BMI quartile both in OCn and ucOCn. No significant changes were found in total OC mean values among BMI quartiles in either PreM or PostM (From the 1st to 4th quartiles: PreM: 37.5±3.0; 33.9±9.9; 27.2±12.2 and 25.1±17.4 ng/ml; PostM: 35.9±13.2; 43.6±21.5; 28.5±12.8; 28.3±16.3). In both menopausal status, the mean OCn levels presented a tendency to have lower values in the 3rd and 4th quartiles as compared to the 1st and 2nd. Conversely, there was a significant increase in ucOC mean levels as BMI increase, both in PreM and in PostM groups (from 1st to 4th quartiles: PreM: 1.53±1.08; 1.93±1.25; 3.25±1.72 and 4.27±0.86 ng/ml; ANOVA P=0.0003, Tuckey post‐test: P<0.01 between 1st and 4th and between 2nd and 4th quartiles; PostM: 1.09±1.04; 1.88±1.62; 2.74±1.51; 4.21±1.83; ANOVA P=0.0009, Tuckey post‐test: P<0.001 between 1st and 4th and P<0.05 between 2nd and 4th). Leptin levels also increased with BMI in both PreM and PostM.

Conclusion

The present results confirm that BMI presents a significant influence on OC decarboxylation through leptin levels. Moreover, as ucOC is involved in glycemic control, the present results suggest that glucose homeostasis is affected by the increase in BMI.

Support or Funding Information

Supported by Universidad de Buenos Aires, UBACyT 20720130100006BA
Effect of Histone Deacetylase Inhibitors on Vitamin D Dependent Gene Expression in Human Colorectal and Breast Cancers
Authors: Hossain Sharmin
Published: November 2023

There are little evidence on the effect of histone deacetylation by a specific histone deacetylase inhibitor (HDACI), sulforaphane, on vitamin D dependent genes in human colorectal and breast cancers. Sulforaphane is a bioactive compound present in cruciferous vegetables like broccoli, Brussel sprouts etc. In the present study, we examined the influence of sulforaphane on expressions of vitamin D dependent genes, VDR and TRPV6, via inhibition of histone deacetylation with TSA as a positive control. We also examined DKK1, one of the inhibitors of Wnt signaling pathway, to see any potential effect of Vitamin D, sulforaphane (SFN) and/or the combination treatment on its regulation. The effects of SFN and TSA with and without the presence of vitamin D were tested on Caco‐2 and TMX2–28 (breast cancer) cells. Although not statistically significant, we found vitamin D to increase the expressions of VDR (2.3 fold) and TRPV6 (2.8 fold) compared to control. The expression of TRPV6 was increased two‐fold by the combination of vitamin D+TSA and three‐fold by TSA alone. DKK1 expression was increased upon treatment with vitamin D+TSA (fold increase 2.8) and TSA alone (fold increase 3.4). DKK1 expression was also increased by SFN (2.7 fold) suggesting a stand‐alone effect. Expression of DKK1 was also increased in TMX2–28 cells upon treatment with sulforaphane (1.5 fold) and the combination of vitamin D + sulforaphane (1.4 fold). Vitamin D (alone or in combination) with sulforaphane can be an effective nutrigenomic intervention for colorectal and/or breast cancer through the inhibition of histone deacetylation‐ a crucial epigenetic modification process. This research, therefore, has the potential to create a prospective model for epigenetic control in cancer.

Support or Funding Information

Hatch Grant# MAS00992
Muscadine Grape Extract Prevents Proliferation of HER2 Positive Breast Cancer Cells in Association with a Decrease in Phosphorylation of AKT and mTOR
Authors: Mackert Jessica D. , Tallant E. Ann , Gallagher Patricia E.
Published: November 2023

Breast cancer is the most common cancer among American women and the second leading cause of cancer death. Human epidermal growth factor receptor 2 (HER2) positive breast cancer, characterized by overexpression of the HER2 protein, is an aggressive type of breast cancer comprising about 20% of diagnosed cases. Although targeted HER2 drugs are successful in treating HER2 overexpressing breast cancer with up to 70% of patients responding to treatment, less than 10% of the patients achieve total remission and many women become resistant, highlighting the need for additional therapies for this disease. Grape seed and/or skin extracts are popular dietary supplements due to their high polyphenol content and potent anti‐oxidant and anti‐inflammatory properties. Recently, numerous studies showed the cancer chemotherapeutic potential of extracts prepared from or active components isolated from a number of different species of grapes. The muscadine grape (Vitis rotundifolia), found in the warm, humid climate of the southeast United States, has a distinct phytochemical composition as compared to other grape varieties. The muscadine grape contains a high concentration of anthocyanin 3,5‐diglucosides, ellagic acid, ellagic acid precursors, gallic acid, flavan‐3‐ols and flavonols. Extracts from muscadine grapes have not been extensively studied and information regarding their therapeutic effects on cancer is limited. The objective of this study was to determine whether a muscadine grape skin and seed extract (MGE) inhibits the proliferation of HER2 positive breast cancer cells. Treatment with increasing concentrations of MGE (a proprietary MGE from Piedmont Research & Development Corp.; 10 – 40 μg/mL) significantly inhibited proliferation of human SKBR3 HER2 overexpressing breast cancer cells. The reduction in growth was dependent upon the dose of the MGE with an observed maximal effect at 40 μg of phenolics/mL (58.4% inhibition; n=3, p < 0.0001). In addition, incubation of human SKBR3 cells with 30 μg/mL of MGE attenuated cell proliferation in a time‐dependent manner with a maximal 49.2% reduction after 48 h (n=3; p < 0.0001). Protein kinase B (AKT) and its downstream effector, mechanistic target of rapamycin (mTOR), are implicated in the regulation of breast cancer proliferation and cell survival. Treatment of SKBR3 cells with MGE (20 μg of phenolics/mL) for 16 h significantly reduced phospho‐AKT by approximately 70% (n=4, p ≤ 0.01) and phospho‐mTOR by greater than 50% (n = 5, p ≤ 0.05) compared to untreated SKBR3 cells. These results suggest that MGE attenuates proliferation in HER2 positive breast cancer cells by reducing phospho‐AKT which inhibits phosphorylation of mTOR. Thus, MGE may serve as an effective therapeutic either administered singly or in a combination regimen with targeted drugs for the treatment of HER2 positive breast cancer.
A Western Dietary Pattern Has The Highest Environmental Footprints Among Lebanese Adults: Findings From A Cross Sectional Study Among Lebanese Adults
Authors: Naja Farah , Jomaa Lamis , Itani Leila , Zidek Jeremy , Hwalla Nahla
Published: November 2023

Background

The recently released Sustainable Development Goals call for sustainable food consumption. Dietary patterns and Food‐Based‐Dietary‐Guidelines are being revisited and studied in terms of their sustainability in addition to their health implications. In this study we aim to evaluate and compare the Environmental Foot Print (EFP) of dietary patterns, including the Western and the Lebanese‐Mediterranean, prevalent among Lebanese adults.

Methods

Dietary patterns previously derived in a sample of Lebanese adults (n=337) included: Western, Lebanese‐Mediterranean and High‐Protein. In this study, these patterns were examined for their EFP including soil erosion, water use, energy use, Greenhouse Gas (GHG) emissions and economic cost. Metrics for these EFP were calculated using review of life cycle analyses (LCAs) on a functional unit of 1 kg of food consumed and analyzing the food supply chain from agriculture production to household consumption, including food waste. Whenever available, national and regional estimates were used. Retail food costs were obtained from cost of living statistics or listed prices at local grocery stores.

Results

Results of the multiple logistic regression showed that, compared to the Lebanese‐Mediterranean and the High‐Protein dietary patterns, a one‐unit increase in the previously derived Western pattern score led to highest odds of belonging to the third tertile of metrics for soil erosion (2.33 vs 1.56 and 1.38), water use (3.23 vs 1.26 and 1.73), GHG emissions (4.03 vs 1.78 and 2.81), and cost (3.51 vs 1.55 and 1.75). Reported odds are significant at p<0.05. Older age (>40 years) and females had lower estimates of GHG emissions and cost, while a university education level had lower values for water and energy use. Smokers had significantly higher values for the five EFP metrics.

Conclusions

Findings of this study showed that, of dietary patterns prevalent among Lebanese adults, the Western had the highest EFP. Coupled to our earlier findings of this pattern's deleterious effects on obesity, diabetes, and metabolic syndrome, this study lends evidence for the notion that what is unhealthy for people may also be unhealthy for ecosystems and underscores the importance to develop evidence‐based interventions to halt the nutrition transition which Lebanon and other countries around the globe are witnessing.

Support or Funding Information

None
Crosstalk between Lactobacillus reuteri and mammalian intestinal epithelium
Authors: Gsnesh Bhanu Priya , Hall Anne , Ayyaswamy Sriram , Fultz Robert , Versalovic James
Published: November 2023

Lactobacillus reuteri ATCC PTA 6475, probiotic bacterium, with an intact hdc gene cluster is known to synthesize and secrete histamine, and can suppress inflammation in mammalian systems via specific activation of the type 2 histamine receptor (H2R). However, it is unclear how L. reuteri‐derived histamine avoids activation of inflammatory H1 receptors in the intestinal epithelium. In this work, we identified a soluble secreted isoform of diacylglycerol kinase (Dgk) from L. reuteri 6475. DGKs belong to a distinct, conserved family of intracellular lipid kinases that phosphorylate diacylglycerol (DAG), catalyzing its conversion into phosphatidic acid (PA). This reaction attenuates DAG levels in the cell membrane, modifying host intracellular signaling proteins that have evolved the ability to bind this lipid for activation. Histamine binding to H1R can cause phosphorylation of PKC via lipid DAG activation. We found that L. reuteri 6475 suppressed basal levels of the pro‐inflammatory cytokines IL‐6, IL‐1, Eotaxin (eosinophilic chemoattractant proteins) and G‐CSF in the luminal mucosa and in blood plasma. L. reuteri lacking dgkA expression could not suppress the aforementioned pro‐inflammatory biomarkers. In addition, we demonstrate for the first time that histamine synthesized by L. reuteri 6475 activates both H1 and H2 receptors, but Dgk synthesized by the bacterium terminates H1R downstream signaling. Inhibition of signaling downstream of H1R was shown by decreased levels of PKC phosphorylation of intestinal epithelium in wild‐type (WT) L. reuteri treated but not in ΔdgkA mutant treated germ‐free (GF) mice. In addition, we also recorded suppression of CD11b+Gr1+ Ly6Ghi immature myeloid cells (IMCs) after WT L. reuteri treatment. In line with mouse experiment PKC phosphorylation was reduced in human epithelial cells after treating the cells with L. reuteri derived conditioned media in vitro. L. reuteri suppress immune response by direct effects of the metabolite histamine and an enzyme diacylglycerol kinase that removes a pro‐inflammatory signal. As a next step we started looking at the effect of L. reuteri and the corresponding mutants in a DSS induced colitis mouse model with conventional microbiome.

Support or Funding Information

This work was supported by the National Institutes of Health (R01 AT004326, UH3 DK083990, and U01 CA170930), the NIH (National Institute for Diabetes and Digestive and Kidney Diseases)‐funded Texas Medical Center Digestive Diseases Center (DK56338), and unrestricted research support from BioGaia AB (Stockholm, Sweden) (J.V.).
Sex Differences in Snack Food Reinforcement in Response to Increasing Dietary Protein
Authors: Casperson Shanon L , Johnson LuAnn , Roemmich James N
Published: November 2023

BACKGROUND

Protein is posited to play a dynamic role in energy balance and reward‐driven eating behavior. However, little is known about the effect of increasing protein intake on snack food reinforcement.

OBJECTIVE

We sought to determine the extent to which increasing dietary protein changes the reinforcing value of snack foods in males and females.

METHODS

In a randomized crossover study, 11 females (age = 25 ± 7y; BMI = 21 ± 2kg/m2) and 10 males (age = 22 ± 2y; BMI = 24 ± 2kg/m2) consumed a standard (15%) and high (30%) protein meal. Study visits were separated by at least one week and menstruating females were studied during the luteal phase (Days 15 – 20). Food reinforcement was assessed using a computer‐based choice task (operant responding with concurrent log2(x) reinforcement schedules) 4h post‐meal. Operant responding was ln‐transformed prior to analysis to reduce skewness. Highly‐palatable, energy‐dense sweet and savory foods, which are most often chosen as snacks, were used to assess food reinforcement. Hedonic (liking) ratings for snack foods used to assess food reinforcement were measured prior to the start of the study via a 10‐point visual analog scale.

RESULTS

Hedonic ratings did not differ between the sexes for the snack foods. Snack food reinforcement was significantly greater in the males than females (p = 0.0477) and sweet snack foods were more reinforcing than savory snack foods (p = 0.0198). There were sex by dietary protein amount (p = 0.0348) and sex by snack food type (p < 0.0001) interactions for snack food reinforcement. Increasing dietary protein reduced food reinforcement, predominantly for the savory snack foods, in the females. Interestingly, food reinforcement in the males was not influenced by dietary protein or snack type. In addition, the reinforcing value of sweet foods did not differ between the sexes and was not influenced by changes in dietary protein.

CONCLUSION

These preliminary results show that increasing dietary protein influences food reinforcement differently in males and females. Understanding the differential effects of meal macronutrient composition on the reinforcing value of foods will provide important insight into psychophysiological mechanisms of snacking behavior.

Support or Funding Information

USDA‐ARS 3062‐51000‐051‐00D
Sustained Postprandial Muscle Protein Synthesis Rates after Protein Ingestion in Healthy Young Males
Authors: Burd Nicholas A. , van Vliet Stephan , van Loon Luc J.C. , Beals Joseph W. , Paluska Scott A.
Published: November 2023

We determined the effect of ingesting a single meal‐like amount of protein on protein digestion and absorption kinetics and the subsequent time‐dependent regulation of postprandial muscle protein synthesis rates in vivo in humans. Seven healthy young males (age: 22±1 y, BMI: 24.8±1.2 kg/m2) received a primed continuous L‐[ring‐2H5]phenylalanine infusions and ingested 38 g intrinsically L‐[1‐13C]phenylalanine‐labeled milk protein concentrate. Biopsies were collected from the vastus lateralis in the basal‐state and 1, 2, 3, and 5 h after protein ingestion. Exogenous phenylalanine appearance in the circulation rapidly increased after protein ingestion (P<0.001). Dietary protein‐derived phenylalanine released in the circulation was 11±2% of the total intake within the first 1 h, followed by 14±1%, 12±1%, and 20±1% within the subsequent 1–2 h, 2–3 h, and 3–5 h of the postprandial period, respectively (total 56±4% over the 5 h period). Myofibrillar protein synthesis rates tended (P=0.10) to increase (absolute change from basal: 0.010±0.008 %/h) immediately following protein ingestion at 0–1 h. Myofibrillar protein synthesis rates were significantly increased at 1–2 h (0.035±0.012 %/h) and myofibrillar protein synthesis remained elevated between 2–3 h (0.019±0.005 %/h) and 3–5 h (0.016±0.004 %/h) after protein ingestion when compared to basal rates (all, P<0.05). Our results show that postprandial muscle protein synthesis rates rapidly increase after protein ingestion and remain elevated above baseline values for at least 5 hours after protein ingestion in healthy young males.

Support or Funding Information

Illinois Campus Research Board
Whey protein protects muscle mass loss of older adults during 7 days of inactivity and accelerates recovery of aerobic fitness during rehabilitation
Authors: Arentson‐Lantz Emily J , Galvan Elfego , Wacher Adam , Paddon‐Jones Doug
Published: November 2023

Older adults are at a greater risk for losing muscle mass and strength during inactivity. Many experience lasting impairments in physical function following an illness or injury that requires hospitalization. We hypothesize that providing older adults with a high quality protein during inactivity will partially preserve muscle mass and function during bed rest and promote recovery during rehabilitation. Up to this point in recruitment, men and women (12 male, 7 female) with an average age of 68.6 years old have been randomized and admitted to the Institute for Translational Sciences' Clinical Research Center for 7 days of bed rest followed by 7 days of inpatient rehabilitation. Dietary intake was strictly controlled and energy intake was calculated for subjects' level of activity. The protein composition of the subjects randomized to the control group (Mixed; n=10) was a mixture of moderate to high quality plant and animal protein. Subjects randomized to the whey group (Whey n=9) received 70% of their protein as whey protein isolate. We are assessing body composition using iDEXA, measuring muscle strength using isokinetic dynamometry, aerobic capacity via VO‐2‐ max testing. Preliminary findings indicate that consuming whey may slow the loss of lean leg mass (−1035 ± 167 vs. −709 ± 101 g for Mixed vs Whey), promote fat mass loss (−14 ± 92 vs. −407 ± 155 g for Mixed vs. Whey) during inactivity. Furthermore, consuming whey protein during rehabilitation accelerates recovery of relative VO2 max (−8.9 ±3.6 vs. 0.41 ± 3.65 ml/kg/min for Mixed vs. Whey).

Support or Funding Information

This research was supported by: Dairy Research Council, RO1 NR012973, and P30 AG024832.
Plasma Glutamine is a Minor Precursor for the Synthesis of Citrulline: a Multispecies Study
Authors: Marini Juan , Agarwal Umang , Didelija Inka C. , Azamian Mahshid S. , Stoll Barbara , Nagamani Sandesh C.S.
Published: November 2023

Glutamine is considered the main precursor for citrulline synthesis in many species, including humans. The transfer of 15N from 2‐[15N] glutamine to citrulline has been used as evidence for this precursor‐product relationship. Research in mice, however, has shown that nitrogen and carbon tracers follow different moieties of glutamine and that glutamine contribution to the synthesis of citrulline is minor. It is unclear if this small contribution of glutamine is also true in other species, including humans. For this reason the current research was designed to determine the contribution of glutamine to citrulline production using nitrogen and carbon skeleton tracers in multiple species. Humans (n=4), pigs (n=5), rats (n=6), and mice (n=5), were infused with L‐2‐[15N] and L‐[2H5] glutamine, and L‐5,5‐[2H2] citrulline for 4 h. The contribution of glutamine to citrulline synthesis was calculated using different precursor‐product pairs: ‘glutamine M+1 to citrulline M+1’, ‘2‐[15N] glutamine to 2‐[15N] citrulline’ and ‘[2H5] glutamine to [2H5] citrulline’. Species specific differences in glutamine and citrulline fluxes were found (P < 0.001), with rats having the largest fluxes, followed by mice, pigs and humans (glutamine: 3789±162, 3228±89, 882±39, 423±25 μmol•kg−1•h−1 and citrulline: 198±11, 81±5, 47±3, 11±2 μmol•kg−1•h−1, respectively). The contribution of glutamine to citrulline estimated using ‘glutamine M+1 to citrulline M+1’ ranged from 88±8% in humans to 46±3% in pigs (rats= 57±3%; mice= 60± 4%). However, using 2‐[15N] glutamine and 2‐[15N] citrulline as precursor and product yielded values of 48±5% in humans and 28±2% in pigs (rats=35±3%; mice= 29± 4%). Furthermore, the use of ‘[2H5] glutamine to [2H5] citrulline’ yielded lower values (P < 0.001), resulting in a contribution of glutamine to the synthesis of citrulline of 10±1% in humans and 3±0.4% in pigs (rats=17±1%; mice= 14± 1%). In conclusion, the use of [15N] glutamine to determine the contribution of glutamine to the synthesis of citrulline yields higher values than the use of the deuterated tracer. This is due to the recycling of the 15N label and the non‐specific incorporation into any of the three nitrogen groups of citrulline. In contrast, the deuterated tracer follows the carbon skeleton of glutamine and for this reason provides a truthful estimate of the contribution of glutamine to the synthesis of citrulline. This work shows that glutamine is a minor precursor for the synthesis of citrulline in humans, pigs, rats and mice.

Support or Funding Information

NIH R01 GM108940; NIH R03 HD078447; USDA/ARS 58‐3092‐5‐001
HDL‐Associated Proteins and LDL Differentially Modulate Chronic Myelogenous Leukemia Cell Viability
Authors: Andersen Catherine J. , Murray Kristina , Ragonesi Nicholas , Doerr Adam , Cintron‐Rivera Layra , Dupree Lydia
Published: November 2023

HDL has been shown to exert anti‐cancer effects. One of the mechanisms by which HDL impairs cancer cell growth is through mediation of cellular cholesterol and phospholipid efflux, thereby limiting the availability of essential lipids for rapidly proliferating cells. Cholesterol efflux is facilitated through interactions between cellular ATP‐binding cassette transporter A1 (ABCA1) and apolipopotein A1 (apoA1), in addition to efflux via ABCG1, scavenger receptor class B type 1 (SR‐BI), and passive diffusion to lipidated HDL. Efflux can further be regulated serum amyloid A (SAA), a pro‐inflammatory acute phase protein shown to be elevated in cancer, infection, and obesity. Despite the ability of SAA to displace apoA1 on HDL, studies have reported SAA to both increase and decrease cholesterol efflux. In this study, we investigated the effects of lipoproteins (HDL and LDL) and HDL‐associated proteins (apoA1 and SAA) on chronic myelogenous leukemia cell viability. K562 cells were cultured in the presence of human HDL or LDL (10, 25, or 50 ug/mL) for 24 hours, then assessed for viability. While no changes in K562 cell viability were observed with HDL treatment, LDL dose‐dependently increased cell viability. We further examined the effects of lipoproteins on K562 viability in the presence of the liver X receptor (LXR) agonist TO901317 (5 uM), which is known to induce expression of HDL‐related transporters. Accordingly, we observed significantly increased expression of ABCA1 at 24hr and 72hr following TO901317 treatment. Interestingly, the addition of HDL to TO901317‐treated cells did not affect K562 cell viability, whereas LDL dose‐dependently increased viability. Conversely, treatment of cells with human‐derived apoA1 or HDL + apoA1 drastically reduced K562 cell viability after 48hr and 72hr. K562 cell viability was modestly decreased by SAA treatment, with greater decreases observed in cells treated with SAA + HDL. These findings suggest that HDL‐associated proteins and LDL differentially regulate chronic myelogenous leukemia cell viability

Support or Funding Information

The Science Institute of the College of Arts and Sciences and Faculty Research Committee, Fairfield University
Effects of medium chain fatty acid on the expression of the genes related with glycolytic and fatty acid synthetic pathways in the mice refed after starvation
Authors: Takami Saeko , Matsumoto Kineko , Honma Kazue , Mochizuki Kazuki , Goda Toshinao
Published: November 2023

Medium chain fatty acid (MCFA) is absorbed without a micelle formation, and transferred via portal vein to the liver, where it is used as an efficient energy source. Thus, medium chain triacylglycerol (MCT) is useful for energy supple in the postoperative patients with reduced digestive functions. In this study we have examined whether refeeding a diet containing MCT after starvation in mice affects the expression of the genes related with glycolytic and fatty acid synthetic pathways in the liver. Adult C57BL/6J mice were fasted for three days, and refed a liquid diet containing either 27% of energy as long chain triacylglycerol (LCT), 21% as LCT and 6% as MCT, or 9% as LCT and 18% as MCT, along with 40% of energy as carbohydrate and 22% as protein. The mice were given the diets through a gastric tubing three times every 4 h (12 kcal/100 g BW/12 h). The mice were killed at 12 h after refeeding, and the serum and the liver were collected for determinations of triacylglycerol concentrations. The mRNA levels of the genes related with glycolytic and fatty acid synthetic pathways were determined by RT‐PCR. Triacylglycerol concentrations in the serum and the liver were similar among the three refed groups. The mRNA levels of glucokinase and fatty acid synthase genes in the liver were significantly higher in the mice refed LCT diet than those in starved mice, and they tended to be lower in the mice refed MCT diet. The mRNA level of carbohydrate‐responsive element‐binding protein (ChREBP) gene was significantly greater in the mice refed LCT diet than that in starved mice, but they were not significantly elevated in the mice refed MCT diet. The results suggest that refeeding a diet containing MCT after fasting leads to a reduced expression of ChREBP gene, which is likely to be involved in the suppression of the genes related with glycolytic and fatty acid synthetic pathways.
The Role of FADS1 and FADS2 Genes in Oleic Acid (18:1n‐9) Metabolism: FADS1 Δ7‐Desaturates Gondoic Acid (20:1n‐9, 11–20:1) to 7,11–20:2
Authors: Park Hi Kyu , Engel Matthew G. , Vogt‐Lowell Kyle , Lawrence Peter , Kothapalli Kumar , Brenna J. Thomas
Published: November 2023

Introduction

In mammals, FADS2 catalyzes “front‐end” Δ4‐, Δ6‐, and Δ8‐desaturation of fatty acyl chains, whereas FADS1 has Δ5‐desaturase activity. 18‐ and 20‐carbon precursors of highly unsaturated omega‐3 and omega‐6 fatty acids are common substrates for FADS1 and FADS2.

Our main objective was to characterize the metabolic fate of oleic acid (OA) by FADS gene product.

Methods

MCF‐7 cells were stably transformed with either FADS1 or FADS2 via pcDNA3.1 expression vector. A series of dose‐response experiments were conducted with albumin‐bound fatty acid substrates (18:1n‐9 and 20:1n‐9) provided in concentrations ranging from 0–100 μm. Cells were harvested after 24 h, fatty acid methyl esters were analyzed by GC‐FID and structures were confirmed by GC‐covalent adduct chemical ionization tandem mass spectrometry (GC‐CACI‐MS/MS).

Results

When stable cells were incubated with 18:1n‐9, FADS1 and control cells elongated 18:1n‐9 ‐> 20:1n‐9, and FADS2 cells Δ6‐desaturated, elongated and then Δ5‐desaturated via FADS1‐coded activity in MCF7 cells: 9–18:1 ‐> 6,9–18:2 ‐>8,11–20:2 ‐>6,8,11–20:3 (20:3n‐9). Surprisingly, FADS1 cells Δ7‐desaturated 20:1n‐9 ‐> 7,11–20:2. Our results imply two pathways for oleic acid metabolism: 18:1n‐9 ‐> (Δ6‐desaturation) ‐> 18:2n‐9 ‐> (Elongation) ‐> 20:2n‐9 ‐> (Δ5‐desaturation) ‐> 20:3n‐9 and 18:1n‐9 ‐> (Elongation) ‐> 20:1n‐9 ‐> (Δ7‐desaturation) ‐> 7,11–20:2.

Conclusion

Alternative pathways for oleic acid metabolism exist, with the metabolic fate of OA dependent on the relative FADS2 or elongase activities. Additionally, we present the first evidence of Δ7‐desaturation via the FADS1 gene product.

Support or Funding Information

NIH R01 AT007003
A Need to Analyze Radiolabeled Cholesteryl Ethers for Biological Stability before Use
Authors: Manual Denny Joseph Kollareth , Hansen Inge Holm , Deckelbaum Richard Joseph
Published: November 2023

Background

Radiolabeled cholesteryl ethers are widely used as nonhydrolyzable tracers to label lipid emulsions and lipoproteins in a variety of in vitro and in vivo investigations. The stable ether bond is not cleaved by mammalian cellular enzymes, and ethers do not readily escape from cells. Hence, radiolabeled cholesteryl ethers accumulate in cells and organs to provide a cumulative marker of the uptake of emulsions and lipoproteins.

Aim

To compare the biological stability of two commercially available [3H]‐cholesteryl ethers: [3H]‐cholesteryl ether‐A (company A); [3H]‐cholesteryl ether‐ B (company B).

Methodology

Biological stability of cholesteryl ether was determined in both in vitro and in vivo models using triglyceride (TG) emulsions labeled with [3H]‐cholesteryl ether. J774 A2 macrophages were incubated for 4 & 24 hrs with commercially available Intralipid labeled with [3H]‐cholesteryl ether‐A or [3H]‐cholesteryl ether‐B. Further, neonatal mice were intraperitoneally injected with laboratory made TG emulsions of docosahexaenoic acid (tri‐DHA) labeled with [3H]‐cholesteryl ethers. Lipids were extracted and hydrolysis of cholesteryl ethers were determined by radioactivity in the cholesteryl ester (CE) and free cholesterol (FC) bands after separation by thin layer chromatography (TLC).

Results

On TLC, 87% of [3H]‐cholesteryl ether‐A was found at the TG region, while 96% of [3H]‐cholesteryl ether‐B was found at the CE region. Among cells exposed to Intralipid labeled with [3H]‐cholesteryl ether‐A, 14% of the label was found as FC at 4 hrs and 42% was found as FC at 24 hrs. Among cells exposed to Intralipid labeled with [3H]‐cholesteryl ether‐B, 93% of the label was in unhydrolyzed form at both 4 & 24 hrs. Among neonatal mice administered [3H]‐cholesteryl ether‐A labeled tri‐DHA emulsion, 48 % of the label was found as FC in liver at 4 & 24 hrs, while the label was found to be unhydrolyzed among animals injected with [3H]‐cholesteryl ether‐B labeled tri‐DHA emulsion.

Conclusions

[3H]‐cholesteryl ether‐A was catabolized to FC both in vitro and in vivo. FC, unlike cholesteryl ether, can then reenter the circulation leading to false results. Radiolabeled cholesteryl ethers need to be analyzed for biological stability before utilizing them for in vitro or in vivo experiments.
Glycovariations in Key HDL‐Associated Glycoproteins Differentiate Between Clinical Groups and Affect the Immunomodulatory Capacity of HDL
Authors: Zivkovic Angela M , Krishnan Sridevi , Shimoda Michiko , Sacchi Romina , Muchena John K , Luxardi Guillaume , Kaysen George A , Parikh Atul N , Ngassam Viviane , Johansen Kirsten , Chertow Glenn , Grimes Barbara , Smilowitz Jennifer T , Maverakis Emanual , Lebrilla Carlito B
Published: November 2023

The goal of this pilot study was to determine whether HDL composition, particularly key HDL‐associated glycoproteins, affects HDL immunomodulatory function. HDL were purified from healthy controls (n=13), subjects with metabolic syndrome (MetS) (n=13), and diabetic hemodialysis (HD) patients, who experienced an infectious hospitalization event within 60 days (HD+) (n=12), and those with no event (HD−) (n=12). Concentrations of HDL‐bound serum amyloid A (SAA), lipopolysaccharide binding protein (LBP), apolipoprotein A‐I (ApoA‐I), apolipoprotein C‐III (ApoC‐III), α‐1‐antitrypsin (A1AT), and α‐2‐HS‐glycoprotein (A2HSG); and the site‐specific glycovariations of ApoC‐III, A1AT, and A2HSG were measured. Secretion of interleukin 6 (IL‐6) in stimulated monocytes was measured to assess HDL immunomodulatory capacity. HDL from HD patients were enriched in SAA, LBP, ApoC‐III, disialylated ApoC‐III (ApoC‐III2) and desialylated A2HSG. HDL that increased IL‐6 secretion were enriched in ApoC‐III, disialylated glycans 5402 and 5412 at multiple A1AT glycosites and desialylated A2HSG, and depleted in monosialylated ApoC‐III (ApoC‐III1). HDL from HD+ patients were enriched in SAA but had lower levels of sialylation across glycoproteins. Our results demonstrate that HDL composition, including the site‐specific glycosylations, differentiate between clinical groups, correlate with HDL's ability to modulate LPS‐induced monocyte IL‐6 response, and may be predictive of HDL's ability to protect from infection.

Support or Funding Information

This research project was supported by NIH R01GM049077 (CBL), the UC Davis Clinical and Translational Research Center NIH grant # UL1 TR000002 (AMZ), and by the Burroughs Wellcome Fund and a grant from the NIH 1DP2OD008752 (EM) contracts N01‐DK‐7‐0005 and N01‐DK‐7‐5004 from the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) and a K24DK085153 grant.
Biomarkers of food intake and nutrient status are associated with glucose tolerance status and development of Type 2 diabetes
Authors: Savolainen Otto , Lind Mads Vendelbo , Bergström Göran , Fagerberg Björn , Sandberg Ann‐Sofie , Ross Alastair
Published: November 2023

BACKGROUND

Diet is frequently associated with both the development and prevention of type 2 diabetes (T2D) but there are a lack of objective tools for assessing the causal relationships between diet and T2D. Biomarkers of dietary intake could help strengthen the link between a healthy diet and prevention of diabetes.

OBJECTIVE

The objective of this study was to explore how diet is related to glucose tolerance status (GTS) and future development of T2D irrespective of metabolic syndrome (MetS) risk factors, using dietary biomarkers as an objective measure of dietary intake unconfounded by recall and reporting bias.

RESEARCH DESIGN AND METHODS

Dietary biomarkers were measured in plasma from 64‐year old women with different glucose tolerance classifications (normal glucose tolerance; NGT (n=190), impaired glucose tolerance; IGT (n=209), and diabetes (n=230)), randomly selected from the population register in Gothenburg, Sweden. The same subjects were followed up after 5 years to determine changes in glucose tolerance (NGT (n=167), IGT (n=174) and diabetes (n=159)). Analysis of covariance (ANCOVA) adjusted for significant measures of MetS was used to explore baseline data for associations between dietary biomarkers, GTS and new T2D cases at follow up (n=69).

RESULTS

After adjustment for MetS risk factors, alpha‐tocopherol, alkylresorcinols C17 and C19 (markers of whole grain wheat and rye), b‐alanine (meat), eicosapentaenoic acid (fish) and linoleic acid were associated with GTS and 3‐carboxy‐4‐methyl‐5‐propyl‐2‐furanpropanoic acid (CMPF) (fish) and alpha‐tocopherol with future development of T2D.

CONCLUSIONS

Several dietary biomarkers were strongly associated with GTS irrespective of MetS factors, underlining the role of diet in development and prevention of T2D. The use of multiple dietary biomarkers can provide a link with diet that is unencumbered by recall bias normally associated with dietary studies and allows examination of the role of diet even when dietary information is not available.
A Role of IL‐21 in Pulmonary Fibroblast Activation
Authors: Sathiaseelan Roshini , Huang Chaoqun , Senavirathna Lakmini Kumari , Liu Lin
Published: November 2023

Idiopathic pulmonary fibrosis (IPF) is a progressive, fatal interstitial fibrotic lung disease with a poor prognosis. Activated fibroblasts produce excess extracellular matrix leading to fibrosis. Transforming growth factor β (TGFβ) is increased in IPF and is one of the most important factors for the activation of fibroblasts. T lymphocytes‐derived cytokines such as IL‐13 and IL‐4 is known to stimulate macrophages to secrete TGFβ. However, the direct effects of these cytokines on the activation of fibroblasts are poorly understood. In this study, we investigated the role of IL‐21 in pulmonary fibroblast activation. RNA sequencing analysis revealed that TGFβ1 increased the expression of IL‐21 receptor (IL‐21R) in primary human pulmonary fibroblasts (HPF). The up‐regulation of IL‐21R was confirmed by real‐time PCR in normal pulmonary fibroblasts (HPF, CCD13, CCD8) and IPF fibroblasts (LL97A and LL29). To determine the effects of IL‐21 on the fibroblast functions, pulmonary fibroblasts were serum‐starved for 16 hours and then treated with TGFβ1 and/or recombinant human IL‐21 for 2 days. The mRNA expression of myofibroblast marker, α‐smooth muscle actin was markedly increased by TGFβ and IL‐21 in all of the fibroblast cell lines. We conclude that IL‐21 promotes the differentiation of fibroblasts to myofibroblasts via TGFβ‐induced IL‐21R expression.

Support or Funding Information

R01HL116876 and P20GM103648
Capsaicin Synergizes with Camptothecin to show Enhanced Apoptotic Activity in Small Cell Lung Cancer
Authors: Friedman Jamie Rae , Perry Haley E , Brown Kathleen C , Akers Austin T , Nolan Nicholas A , Rollyson William D , Chen Yi Charlie , Denning Krista L , Brown Linda G , Dasgupta Piyali
Published: November 2023

Platinum‐based chemotherapy is the cornerstone of small cell lung cancer (SCLC) treatment. Chemotherapy initially shows excellent outcomes with 60–80% patients achieving remission. However, patients relapse within a year and after that the tumor does not respond to chemotherapy or radiation. There is only one FDA‐approved drug for relapsed SCLC, which is camptothecin (topotecan). It has been shown to display an objective response rate of about 3%. Therefore agents which can increase the response rate of camptothecin should be of considerable benefit to SCLC patients. We tested the combinatorial effects of the nutritional compound capsaicin with camptothecin in human SCLC. Caspase‐3 activity assays reveal that the combination of camptothecin and capsaicin displayed greater apoptotic activity than any of the compounds used alone. Statistical analysis using the Chou‐Talalay isobologram showed that capsaicin displayed synergistic apoptotic activity with camptothecin. Chicken chorioallantoic membrane assays confirmed the combinatorial effects of capsaicin and camptothecin. The synergistic effects of capsaicin and camptothecin are mediated by the calpain pathway and involve the proteolytic cleavage of the pro‐apoptotic protein Bax to generate a more potent pro‐apoptotic Bax fragment protein. The results of our studies will foster the hope of novel second‐line treatment regimens for human SCLC.

Support or Funding Information

Funding for our study was supported by an NIH R15‐AREA Grant (2R15CA161491‐02) and a WVU‐MU Health Partnership Grant.
Loss of function mutation in TIMP2 gene accelerates tumorigenesis and mortality in murine model of lung cancer through EGFR signaling
Authors: KUMAR SARVESH , Jensen Sandra , Peeney David , Chowdhury Ananda , Wei Beiyang , Stetler‐Stevenson William G.
Published: November 2023

The aim of this study is to determine the effect of the tissue inhibitor of metalloproteinase‐2 (TIMP‐2) on lung tumorigenesis and investigate the underlying molecular mechanisms using an orthotopic mouse model with a loss of function mutation in the Timp2 gene (T2M). T2M and wildtype (WT) control mice were given 1×106 Lewis lung carcinoma cells transfected with luciferase (LL/2‐Luc‐M38, Caliper) in 50μL PBS via intratracheal installation. IVIS imaging revealed a higher tumor burden in T2M mice compared to WT littermates, suggesting that loss of function of the Timp2 gene enhances tumor growth (p<0.05). We also conducted a Kaplan‐Meier analysis to determine the effect of this mutation on mortality following cancer development. We found that LL/2 tumor‐induced mortality was significantly higher in T2M mice compared to WT controls (p = 0.01). Histologic analysis and H&E staining of lung tissue sections revealed a significant increase in the number of tumor nodules of T2M mice compared to WT controls (p <0.01). CD31 staining showed a significant increase in microvessel density (MVD) in T2M mice compared to WT controls (p<0.01). Given that VEGF is a primary driver of tumor neovascularization, we determined mRNA levels of VEGF expression in healthy and tumor bearing mice by qRTPCR analysis. Interestingly, basal VEGF expression levels were increased in lungs of both non‐tumor bearing and tumor bearing T2M mice compared to WT controls (p<0.01). VEGF is the direct downstream target of HIF‐2a, a transcription factor implicated in tumor hypoxia and expression correlates with decreased overall survival in non– small cell lung cancer (NSCLC) patients. Similarly, an examination of TIMP‐2 and HIF‐2a mRNA expression in a small cohort of patient samples revealed decreased TIMP‐2 and increased HIF2a in NSCLC tumors compared to surrounding non‐tumor lung tissue. Hypoxia induces EGFR signaling through HIF‐2a. Thus, we determined the effect of mutated T2M on the EGFR signaling pathway. We found increased levels of EGFR phosphorylation as well as downstream ERK and Akt activation in tumor bearing and non‐tumor bearing T2M lungs compared to WT controls. In conclusion, these findings offer new avenues for TIMP‐2 research in regulating hypoxic mediators within the tumor microenvironment through EGFR signaling, suggesting TIMP‐2 as a novel bio‐therapeutic for lung cancer therapy.

Support or Funding Information

This work is supported by center for cancer research, intramural research program, NCI/NIH research grants ZIA BC011204 & ZIA SC 009179 to WGSS