bluuebunny/crossref_metadata_2025 huggingface dataset:
The impact of a protein‐rich breakfast on food cravings and reward in overweight/obese ‘breakfast skipping’ adolescent girls
Authors: Ortinau Laura C. , Hoertel Heather A. , Douglas Steve M. , Leidy Heather J
Published: June 2021

Purpose

To assess whether the addition of a high‐protein (HP) or normal‐protein (NP) breakfast alters food cravings and signals controlling food reward in breakfast skipping (BS) girls.

Methods

Using a randomized crossover design, 20 BS girls (age 19±1y; BMI 28.6±0.7kg/m2) where acclimated to each breakfast pattern for 6 days. On day 7, a 4‐h testing day was completed including the consumption of breakfast followed by repeated assessments of food cravings and blood sampling for plasma concentrations of the dopamine metabolite, homovanilic acid (HVA).

Results

Breakfast reduced sweet and savory cravings vs. BS (all, p<0.05). Although no differences in sweet cravings were observed between breakfast meals, HP lead to greater reduction in savory cravings vs. NP (p<0.05). Plasma HVA concentrations were greater following both breakfast meals vs. BS (both, p<0.05), with HP exhibiting sustained increases prior to lunch vs. NP (p=0.09). HVA concentrations were correlated with perceived fullness, breakfast palatability, and protein content at breakfast.

Conclusion

These data suggest that the daily addition of a protein‐rich breakfast, containing 35g of protein, alters signals associated with food motivation and reward, and might be a beneficial strategy to combat the modern food environment in young people.

Grant Funding Source: The Beef Checkoff; Egg Nutrition Center; Margaret Flynn
The short‐term effects of black pepper (Piper nigrum) and rosemary (Rosmarinus officinalis) on sustained attention and energy and fatigue mood states in young adults with low energy
Authors: Lindheimer Jake B. , Loy Bryan D. , O'Connor Patrick J.
Published: June 2021

The aim was to test if black pepper (2 grams) or rosemary (1.7 grams) improved sustained attention, motivation, or feelings of mental energy and fatigue. Outcomes were measured in 40 low‐energy adults before and 60‐ and 90‐minutes after consumption of capsules containing black pepper, rosemary or a placebo (3.1 grams flour). Sustained attention was measured using a 16‐minute dual task in which single digit numbers were presented every second on a screen and the participant performed both a primary task (detection of three successive, different odd digits) and a secondary task (detect the number 6). Feelings of energy and fatigue were measured using the vigor and fatigue subscales of the Profile of Mood States and visual analog scales (VAS). ANOVAs showed non‐significant Condition X Time effects for motivation, also measured with a VAS, and the intensity of energy and fatigue feelings. Time‐varying ANCOVA, controlling for motivation, showed no significant effects on the primary or secondary task outcomes of: correct responses, errors, speed of response and signal detection sensitivity. It is concluded that black pepper and rosemary, consumed in capsule form, in the doses used and while wearing a nose clip to block olfactory effects, do not induce a short‐term improvement in sustained attention, motivation to perform cognitive tasks, or feelings of mental energy and fatigue in young adults with low energy.
Methylmalonic acid concentration in dried blood spots – validation of a novel method for vitamin B12 status assessment
Authors: Schroder Theresa H. , Quay Teo A. W. , Lamers Yvonne
Published: June 2021

Marginal vitamin B12 (B12) deficiency has been associated with an increased risk of adverse pregnancy outcomes and cognitive decline. About 10 % of the US population and 25 % of the Canadian population show inadequate B12 status. Methylmalonic acid (MMA) is a functional biomarker for B12 status and a sensitive indicator for marginal B12 deficiency. To date, MMA concentrations are measured in plasma samples obtained from venipuncture. Dried blood spots (DBS) allow non‐invasive, simple collection of biological samples with no need of immediate processing or special storage.

The goal of this study was to establish an analytical method for quantification of MMA in DBS and to validate its application for B12 status assessment. Using liquid chromatography ‐ tandem mass spectrometry, we developed a simple, low‐cost assay that shows high imprecision (CV < 1.2 %) and good recovery (95 %). In 73 healthy female subjects (19 – 30 y), mean (± SD) concentrations of plasma MMA and DBS MMA were 163 ± 65 nmol/L (74 – 458 nmol/L) and 36.4 ± 11 pmol/DBS, respectively. There was a strong correlation between plasma and DBS MMA concentrations (R = 0.90 P < 0.001).

This novel method was shown to be sensitive, reliable, and convenient. MMA analysis in DBS has the potential to be a more practical alternative for B12 screening and to facilitate B12 status assessment in large‐scale population‐based surveys and in populations living in less accessible areas.

Grant Funding Source: ‘FNH Vitamin Research Fund’ University of British Columbia
Comparison of hypolipidemic effects of three berries in diet‐induced obese C57BL/6J mice
Authors: Kim Bohkyung , Lee Sang Gil , Ku Chai Siah , Park Youngki , Yang Yue , Pham Tho X , Wegner Casey , Koo Sung I , Chun Ock K , Lee Jiyoung
Published: June 2021

Berry polyphenols have various health‐promoting properties including antioxidant, anti‐inflammatory, and hypolipidemic effects, which contribute to lowering cardiovascular disease risk. We compared the lipid‐lowering effect of three berries using male C57BL/6J fed a high fat diet (HF, 35% fat by weight, 60% energy from fat) to induce obesity. The mice were fed a HF control diet or a HF supplemented with 5% of freeze‐dried blueberry (BB), 6.3% blackberry (BK) or 5.7% blackcurrant (BC) to have a similar anthocyanin content for 12 wk. BK‐fed mice showed significantly lower plasma triglyceride (TG) concentrations than control, BB and BC groups with no differences in plasma total cholesterol. Plasma antioxidant status (ferric reducing ability of plasma and superoxide dismutase levels) and plasma levels of TNFα, glucose and insulin were not significantly different between groups. mRNA abundance of lipogenic genes (e.g., fatty acid synthase, stearoyl coA desaturase 1, sterol regulatory element binding protein 1c) in the liver and epididymal adipose tissue were not significantly different between control and BK groups. However, there was a trend toward an increase in carnitine palmitoyltransferase 1α in the livers of BK‐fed mice. The results suggest that the hypotriglyceridemic effect of BK is likely due to increased mitochondria fatty acid β‐oxidation in HF‐fed obese mice. (*Equally contributed to this study)
Regulation of genes involved intestinal cholesterol metabolism by polyphenol‐rich black currant extract in Caco‐2 cells
Authors: Kim Bohkyung , Perkins Alyssa , Lee Jiyoung
Published: June 2021

We investigated the effect of blackcurrant extract (BCE) containing 25% polyphenols on the regulation of genes involved in intestinal cholesterol metabolism with a specific focus on transintestinal cholesterol efflux (TICE). As TICE provides a non‐biliary route for cholesterol excretion, TICE activation can lower plasma cholesterol levels. Caco‐2 cells, a human intestinal cell line, were incubated with BCE (50 or 100 μg/mL) for 24 h for gene expression analysis. BCE significantly decreased the expression of genes for cholesterol synthesis (3‐hydroxy‐3‐methylglutaryl coenzyme A reductase and sterol regulatory element binding protein 2), for apical cholesterol uptake (Niemann‐Pick C1 Like 1), and for basolateral cholesterol efflux (ATP binding cassette transporter (ABC) A1). Decreases in the protein levels of these genes were confirmed by Western blot analysis. In contrast, BCE significantly increased mRNA and protein levels of ABCG5/G8, which facilitate apical cholesterol efflux to the intestinal lumen. Furthermore, BCE markedly increased protein levels of LDL receptor. Our data suggest that BCE may activate TICE pathway by increasing LDL uptake into enterocytes and by facilitating the efflux of LDL‐derived cholesterol to the luminal side via increased ABCG5/G8 expression in the intestine.

Grant Funding Source: College of Agriculture and Natural Resources at UCONN and National Research Foundation of Korea
Effect of rosemary supplementation on oxidative stress after myocardial infarction in rats
Authors: Rafacho Bruna Paola Murino , dos Santos Priscila Portugal , de Freitas Gonçalves Andrea , Candido Renata , Fernandes Ana Angélica Henrique , Schmidt Paula Azevedo , Zornoff Leonardo Antonio Mamede , Campana Álvaro Oscar , Minicucci Marcos Ferreira , Paiva Sergio Alberto Rupp
Published: June 2021

Objective

To analyze the influence of rosemary (R) consumption on cardiac oxidative stress after myocardial infarction (MI).

Material

Male Wistar rats weighing 200g were allocated into 6 groups: 1) Sham group fed standard chow (SA0, n = 6); 2) sham group fed standard chow supplemented with 0.02% of dried R (diet 1) (SA1, n = 6); 3) sham group fed standard chow supplemented with 0.2% of dried R (diet 2) (SA2, n = 6); 4) MI rats fed standard chow (IA0, n = 6); 5) MI rats fed diet 1 (IA1, n = 6); 6) MI rats fed diet 2 (IA2, n = 6). After three months of treatment, heart tissue was collected and tested for lipid hydroperoxide (LH) and antioxidant enzymes catalase, gluthatione peroxidase (GPx), superoxide dismutase (SOD) and Nrf2 expression. Comparisons were made by two‐way ANOVA test and post‐test of Holm‐Sidak. Values were presented as mean ± standard error.

Results

MI rats showed higher cardiac levels of LH and SOD than Sham rats. Rosemary supplementation reduced LH with diet 2 and SOD with both doses. No differences were observed for catalase, GPx and Nrf2.

Conclusion

Chow supplementation with rosemary reduced oxidative stress after myocardial infarction. Financial support: FAPESP.
Health and Functional Characteristics of Indigenous Food in the Korean Longevity Belt
Authors: Lee Mee sook , Moon Kwang‐Hyun
Published: June 2021

This study was performed to discover the factors of indigenous food for health and longevity in the Korean longevity belt. The research method was based on the examination of individual cases through in‐depth interviews with 96 subjects, aged 65–102 years old. In Korea, the Kugoksundam longevity belt, their main dietary styles were similar to Korean villages consisting of rice, soup, kimchi and side dishes. However, they consumed a great variety of wild herbs and ripened wild seeds through cultivating and gathering wild types of edibles. The cold side dishes (Namools) include plenty of seasonal wild plants and herbs, and the portion of dried or preserved wild herbs outnumbered any other kinds of food stuff. There were more than 60 kinds including acorns, pine, Zanthoxylum piperitum, bamboo seeds, arrowroot starch, mugwort, sedum, shepherd's purse, sowthistle, aster scaber, Hemerocallis fulva, Ligularia stenocophala, Viola mandshurica, chard, mallow, dandlion, mulberry leaves, japanese ginger, aster romena, royal fern, Scilla scilloides, etc. In the longevity belt these plants were consumed appropriately to exert their tonic properties. These wild herbs appeared to provide a variety of functionality such as anti‐oxidative, anti‐aging, anti‐inflammatory, and anticancer effects through a variety of phytochemicals and fibers contained in these wild plants.
Changes in the metagenome and plasma lipids of hamsters fed high fat diets containing Chardonnay grape seed flour
Authors: Kim Hyunsook , Arvik Torey , Lipson Rebecca , Bartley Glenn E , Kim Dong‐Hyeon , Seo Kunho , Yokoyama Wallace
Published: June 2021

The relationship between changes in the metagenome and cholesterol lowering by defatted chardonnay grape seed were evaluated in male Syrian hamsters fed high‐fat (HF) diets. The diets were supplemented with 10% grape seed flours from chardonnay (CH) or cabernet sauvignon (CS) grapes and compared to a HF diet containing microcrystalline cellulose dietary fiber (control) for three weeks. Hamsters fed the CH diet had significantly lowered plasma total‐, VLDL‐, and LDL‐cholesterol concentrations compared to the control and CS diets. These improved plasma cholesterol changes were correlated with the upregulation of hepatic genes related to cholesterol and bile acid synthesis (CYP51, CYP7A1, and LDLR). Fecal total bacteria concentration was decreased by the CH diet but not significantly (P = 0.056). The CH diet significantly lowered fecal bifidobacterium spp. and lactobacillus spp. concentration compared to control and CS diets. The fecal lactobacillus spp. concentrations from control, CH, and CS groups were significantly positively correlated with plasma total‐ (r=61.6) and LDL‐ cholesterol (r=69.3) concentrations, indicating cholesterol lowering by chardonnary grape seed flour were not due to fecal lactobacillus spp. concentration. Antimicrobial activity of grape seed flour on the intestinal microbiota warrants further investigation.
A solute carrier protein, the mammalian flavonoid transporter, mediates cellular flavonoid uptake
Authors: Lee Ji Hae , Kim Yeojin , Lee Sung‐Joon
Published: June 2021

Flavonoids are phenolic phytochemicals with lipid lowering activities in metabolically active cells. However, cellular flavonoid transport mechanisms are largely unknown. We report that a solute carrier protein, the mammalian flavonoid transporter (MFT), mediates cellular flavonoid uptake. HEK293 cells overexpressing MFT showed significantly increased flavonoid uptake compared with controls. Among tested flavonoids, aglycones were overall better substrates for MFT than glycosides with showing the highest affinity of quercetin to MFT. The uptake of quercetin was inhibited in cells stimulated with a MFT inhibitor and in cells transfected with MFT siRNA. Immunohistochemistry analysis confirmed that MFT is highly expressed in peroxisome where cellular oxidative metabolism is predominant. The hypolipidemic effects of quarcetin were exaggerated in CHO cells overexpressed with MFT compared with control cells. These suggest that MFT could mediate cellular flavonoid uptake, especially in peroxisome, thus its overexpression could enhance hypolipidemic activity of flavonoids.
Comparison and purification of grape pomace extracts for inhibition of intestinal α‐glucosidases
Authors: Kadouh Hoda C. , Sun Shi , Zhou Kequan
Published: June 2021

Alpha‐glucosidases play a major role in controlling starch digestion and therefore postprandial blood glucose, a target for diabetes management. This study aims to prepare and purify active components in grape pomace extract that inhibit intestinal α‐glucosidases. Chambourcin, Merlot, Norton, Petit Verdot, Syrah and Tinta Cão red wine grape pomaces were extracted with 50% acetone and evaluated for rat intestinal α‐glucosidase (25 mg/ml) inhibiting activity. The most potent variety was partitioned into water, butanol and ethyl acetate extracts which were also assayed. The active extract was fractionated via four open column chromatography techniques (C18 and Silica, different eluents) and the retained fractions were tested. At 0.5 mg/ml, Tinta Cão pomace extract was the strongest inhibitor of α‐glucosidase complex (95.34% inhibition, compared to 7.41–88.49%), with its ethyl acetate fraction being the most effective (68.14% inhibition, compared to 9.4–53.4%). Water‐eluted fractions of the preparatory and regular Silica columns outweighed the standard (Acarbose 50 ug/ml, 50% inhibition) at 59.24% and 56.12% inhibition, respectively. Results were significant, suggesting that a potent inhibitor of intestinal α‐glucosidases can be isolated from Tinta Cão grapes for the potential development of a novel antihyperglycemic dietary supplement, following further investigations.

Grant Funding Source: Wayne State University
Protective effect of GSW against UVB‐induced skin damage
Authors: Kim Ok kyung , Ho Jin‐Nyoung , Nam Da‐Eun , Lee Jeongmin
Published: June 2021

Ultraviolet B (UVB) irradiation on skin can induce the production of reactive oxygen species (ROS) which damage cellular components. Increased ROS in skin cause activation of the activator protein‐1 (AP‐1) transcription factor, which increases matrix metalloproteinases (MMPs) expression and collagen degradation. These causes the characteristic changes of photoaging induced by UVB irradiation. In this study, we investigated the protective effects of GSW against UVB induced skin damage in human skin fibroblasts. GSW was first analyzed for antioxidant activity using DPPH and ABTs radical scavenging assay. DPPH and ABTs radical scavenging activities of GSW were significantly higher in a dose‐dependent manner. Skin fibroblast were treated vitamin C and GSW (10μg/mL, 30μg/mL, 50μg/mL) after UVB irradiation (0mJ/cm2 and 25mJ/cm2). Vitamin C and GSW inhibited MMPs (MMP‐1, MMP‐3, MMP‐9) expression and MMP‐1 secretion caused by UVB irradiation. Moreover, we found that treatment with vitamin C and GSW significantly increased type‐1 collagen expression and production. We next examined the levels of antioxidative enzymes (SOD, Catalase, GPx) activities related in antioxidative effects. Reduced antioxidative enzymes activities by UVB irradiation were increased after treatment with vitamin C and GSW. In conclusion, theses results show that GSW has protective effects on UVB‐induced skin damage in human skin fibroblasts by regulating antioxidative defence systems and MMPs expression.
Anti‐obesity effects of Cheonggukjang and Natto extract: from in vivo to in vitro study
Authors: Bae Cho‐Rong , Byun Moon‐Sun , Yu Ok‐Kyeong , Park Tae‐Sun , Cha Youn‐Soo
Published: June 2021

In this study, we investigated the anti‐obesity effects of Cheonggukjang (CK) and Natto (NT) extract on in‐vitro and invivo models. The anti‐obesity effects of CK and NT extract were investigated by measuring Oil Red O staining in 3T3‐L1 adipocytes. Both the extracts reduced the content of Oil Red O stains in 3T3‐L1 adipocytes. For in‐vivo study the C57BL/6 mice were randomly assigned to 5 groups (n=10): Normal diet group (ND), High fat diet group (HD), High fat diet plus soybean (SS), High fat diet plus Cheonggukjang (CK), High fat diet plus Natto (NT). Animals were fed with CK or NT containing experimental diets for 10 weeks, after which they were sacrificed. Bodyweight, bodyweight gain, food intake, energy efficiency ratio, adipose tissue weight, serum insulin, leptin level and lipid profiles along with hepatic mRNA levels of gene involved in fatty acid metabolism were reduced in the CK and NT group. CK and NT groups supplementation modulated mRNA levels of lipolytic genes CPT‐1 and lipogenic genes ACC. These results suggested that the supplementation of Cheonggukjang and Natto have an anti‐obesity effect on high fat diet induced obesity in the mouse model. (This research was supported by the Globalization of Korean Foods R&D program, funded by the Ministry of Food, Agriculture, Forestry and Fisheries, Republic of Korea).
Effect of replacing of pork fat with barley flour in reduced‐fat sausage on cholesterol concentrations in C57BL/6J mice
Authors: Hien Hoang Minh , Lee Sung‐Joon
Published: June 2021

Reduced‐fat sausages were prepared by replacing pork back fat with barley flours at levels of 9% (SS‐9) and 11% (SS‐11; w/w). The effects of these sausages on physical, sensor properties, and lipid metabolism in C57BL/6J mice were investigated. SS‐9 and SS‐11 showed similar textural and sensory properties as the control high fat sausage (HS). Four weeks of feeding, SS‐9 and SS‐11 showed hypocholesterolemic effects with a significant reduction in total cholesterol and induction of high‐density lipoprotein cholesterol compared to the HS group. Additionally, SS‐11 reduced triglyceride and fecal cholesterol and increased fecal bile acids. The mRNA expressions of hepatic CYP7A1 were significantly induced in the SS‐9 and SS‐11 groups compared to the HS group. The mRNA levels of the LDL receptor were significantly higher, whereas mRNA levels of HMG‐CoA reductase were lower in the liver of SS‐11‐fed than that of the control mice. Our data suggest that reduced‐fat sausages exerted hypocholesterolemic effects in mice. The suppression of cholesterol biosynthesis and enhancement of LDL uptake and cholesterol degradation may be the major mechanism underlying this effect.
Wheat Bran for Colon Cancer Prevention: Phytochemicals vs. Fiber
Authors: Sang Shengmin , Zhu Yingdong , Soroka Dominique N
Published: June 2021

Many studies have found that dietary cereal fibers protect against colon cancer, however, not all cereal fibers are equally effective. Wheat bran is the only cereal bran that shows consistent protection against colon cancer in laboratory animal models, and corn and oat bran may enhance colon tumorigenesis. It is still unclear whether the active components are phytochemicals or fibers. We and others have found that phytochemicals in wheat bran instead of fiber are the major active components for colon cancer prevention. The inhibitory effect of wheat bran extract on tumorigenesis was observed in our studies in the Apcmin/+ mouse model. Using human colon cancer cells (HCT‐116 and HT‐29) as the guiding assay, we identified 5‐n‐alkylresorcinols (ARs) as the major components in wheat bran for colon cancer prevention. To further study the structure‐activity relationship of ARs, we synthesized more than 20 ARs derivatives. The effects of the length of the alkyl side chain, the substitution pattern of the functional groups on alkyl side chain, the substitution pattern on the benzene ring as well as the underlying molecular mechanism were also studied. The effects of other components in wheat bran, such as sphingolipids and steroids, will also be discussed.
Infant Feding and Morbidity in African HIV‐1‐exposed Uninfected Infants: the Kesho Bora Study
Authors: Bork Kirsten , Cournil Amandine , Cames Cécile
Published: June 2021

This analysis assessed morbidity risks associated with feeding mode in HIV‐1‐exposed uninfected African infants for two age periods (0–3 and 3–6 mo). HIV‐1‐infected women from 5 sites in Burkina Faso, Kenya and South Africa were enrolled during pregnancy and counselled to either breastfeed (BF) exclusively and wean before six mo postpartum or formula‐feed from birth. Infant morbidity was defined in two distinct ways, as ‘trivial’ (i.e. mother‐reported fever, diarrhea, or vomiting since the preceding clinic visit) or as ‘SAE’ (i.e. selected clinical serious adverse events: gastroenteritis, lower respiratory infections, severe malnutrition or death). The analysis (n= 982) was conducted using multiple piecewise exponential regression with adjustment for study site, maternal CD4 cell count at enrollment and cotrimoxazole prophylaxis (CTX). Feeding mode and CTX were time‐dependent variables, defined monthly. Not BF (i.e. never BF or weaned) was associated with increased risks of ‘trivial’ morbidity from 0–3 mo [HR=1.3 (95% CI: 1.1–1.6), P<0.001] but not from 3–6 mo [1.1(0.8–1.5)], and with increased risks of SAE at 0–3 [5.6 (2.9–11.6), P<0.001] and 3–6 mo [2.9 (1.4–5.8), P<0.01]. Partial or predominant BF modes were not associated with significantly increased risks, as compared to exclusive mode. BF prevents serious morbidity and fatal outcomes in African infants up till 6 months.

Funding: ANRS, WHO, EDCTP.
Why does Human Milk Fortification (HMF) increase Urinary F2‐Isoprostanes in the Premature Infant?
Authors: Friel James , Diehl‐Jones Bill , Cai Chenxi , Hossein Zakir , Kim Jongsoo
Published: June 2021

Human milk fortifiers (HMF) are routinely added to mother's milk (HM) for premature infant to make up for potential nutritional deficiencies. We reported data on 65 premature infants (birth weight < 1500 g) who were either fed HM with varying levels of HMF or formula (F) while they were in hospital receiving oral feeds. A significant increase (P<0.05) in urine_F2‐isoprostanes was found as HMF supplementation increased (Pediatr Res. 2011 Feb; 69(2): 160–4). Subsequently we used an in vitro model of Caco‐2 cells with HMF and lipopolysaccharide (LPS) known to induce inflammation in vivo. When paired with HMF, there was a significant increase in IL‐6 secretion above that of LPS alone. A sample of breast milk from our study supplemented with HMF (>; 50%) generated higher levels of intracellular oxidation in intestinal cells, as indicated by the redox‐sensitive dye Dichlorofluorescein (DCF). Furthermore we now have data from DNA Microarray experiments that, in intestinal cells, HMF significantly up regulates a battery of antioxidant response element (ARE)‐driven genes. Additionally, we have pilot data from fetal intestinal cells (FHS) exposed to either human milk (HM) or HM + HMF). Briefly, enterocyte cell cultures were exposed to digested HM +/− HMF. Following 24 hours recovery, RNA was collected, prepared for qRT‐PCR as per the SABiosciences RT2 ProfilerTM array, which screens for cytokines, chemokines, toll‐like receptors (TLRs) and linkers. Of note is the significant up‐regulation of several interleukins (IL1A, IL6, IL10, IL18) or IL18 receptor accessory protein (IL18RAP), and TLR4. Furthermore, an integrin gene (ITGB2) was significantly down‐regulated, which has implications for intestinal barrier function. This data suggests that some HMFs as constituted induce oxidative stress in vitro and in vivo.

Supported by CIHR and MICH.
The relationship of vitamin D status of exercising breastfeeding women on changes in bone mineral density during the first year postpartum
Authors: Sorvillo Andrea , Wideman Laurie , Lovelady Cheryl
Published: June 2021

Vitamin D is needed for bone remolding; however, its relationship with bone mineral density (BMD) during lactation is unclear. The purpose of this study was to determine if serum vitamin D [25(OH)D] concentrations in lactating women were related to BMD changes from 4–52 wks postpartum (PP). At 4 wks PP, women were randomized to exercise (n=18) or control (n=18) groups for 16 wks. All received a year supply of a multivitamin supplement, containing 10 mcg vitamin D. At 4, 20, and 52 wks PP, BMD was measured by DXA; prolactin, estradiol, and 25(OH)D were analyzed by ELISA; and calcium and vitamin D were assessed by 24‐hr diet recalls. Regression was used to assess the association between 25(OH)D at 52 wks PP and changes in BMD, with the covariates prolactin, estradiol, and calcium at 52 wks PP; group; and weight change from 4–52 wks PP. Mean calcium was 1229 ± 444, 1057 ± 365, and 1067 ± 506 mg/d at 4, 20, and 52 wks PP. Mean dietary vitamin D was 5.0 ± 2.6, 4.7 ± 2.6, and 5.2 ± 3.6 mcg/d, respectively. Mean 25(OH)D at 4 and 52 wks PP were 26.4 ± 2.1 and 25.7 ± 1.7 ng/mL, defined as sufficient by IOM (≥ 20 ng/mL). 25(OH)D was not related to changes in BMD. When including all covariates, prolactin was associated with changes in hip and total body BMD (r2=0.25 and 0.43), but not LS BMD (r2=0.16). Results suggest that a dietary intake of 5 mcg/d with a supplement of 10 mcg/d is adequate for maintaining 25(OH)D concentrations and does not affect BMD during lactation. Supported by NC State University Agriculture Research Services
Effects of Antenatal Micronutrient Supplementation on Plasma Protein Profiles in Nepalese Children
Authors: Lee Sun Eun , West Keith P. , Cole Robert N. , Ruczinski Ingo , Schulze Kerry , Yager James D. , Groopman John , Christian Parul
Published: June 2021

Antenatal multiple micronutrient (MN) supplementation in malnourished settings may favor a healthy phenotype in offspring, but mechanisms remain largely unexplored. Plasma proteomics may offer a way to reveal long‐term metabolic effects of early MN exposure. We are applying quantitative mass spectrometry to explore differences in plasma protein abundance in 500 Nepalese children 6–8 yr of age born to mothers who were randomized (100 each) to receive daily supplements of folic acid (FA), iron+FA (IFA), IFA+zinc (IFAZ), multiple MNs, or placebo from 1st trimester to 6 wk postpartum. All tablets also contained vitamin A. Among ~980 plasma proteins measured in ≥ 10% of children, preliminary analysis reveals 9, 10, 8 and 14 proteins were differentially abundant in the FA, IFA, IFAZ and multiple MN vs. control group (all p< 0.01). So far, insulin‐like growth factor‐binding protein 5, ATP‐binding cassette sub‐family A, and eukaryotic translation initiation factor 3 are differentially abundant from controls in 2 nutrient groups. Insulin‐like growth factor 1 was highly abundant in FA, IFAZ, and multiple MNs compared to placebo. Early life micronutrient exposure may influence early childhood plasma protein profiles by presumably interacting with genes regulating growth and development. Supported by the Bill and Melinda Gates Foundation (Grants GH5241 and GH614).

Grant Funding Source: Bill and Melinda Gates Foundation
Maternal obesity and post‐natal high fat diet disrupt hepatic circadian rhythm in rat offspring
Authors: Shankar Kartik , Borengasser Sarah J , Faske Jennifer , Kang Ping , Gomez‐Acevedo Horacio
Published: June 2021

Offspring of obese (Ob) rat dams gain greater body wt and fat mass when fed high fat diet (HFD) as compared to controls. Alterations of diurnal circadian rhythms are known to detrimentally impact metabolically active tissues such as liver. We sought to determine if maternal obesity (MOb) leads to perturbations of circadian rhythm and energy metabolism in offspring liver following a short (2 wk) high fat (HFD) challenge. mRNA expression on PND35 of putative circadian (CLOCK, Reverbα, Bmal1, Cry, Per) and metabolic (PPARα, SIRT1) genes were suppressed following HFD in lean and Ob groups. Further suppression was evident in Ob‐offspring indicating circadian disruption. Using a mathematical model for the circadian clock, analyses of PPARα and SIRT1 temporal changes in relative mRNA abundance revealed exposure to MOb and/or HFD disrupts the circadian rhythm by (i) decreasing mRNA synthesis via a rate limiting activation constant of expression of nuclear Bmal1 and (ii) increasing non‐specific mRNA degradation of both PPARα and SIRT1. HDAC3 a known regulator of circadian rhythmicity and hepatic lipid metabolism, was decreased at 5/6 time points (4 hr intervals) by exposure to HFD and MOb. HFD disrupts core hepatic clock machinery and key regulators of energy metabolism and is further exacerbated by MOb which may increase the risk for harmful adiposity gains in adulthood. Support NIHR01‐DK084225
Development of a piglet model of neonatal systemic Staphylococcus aureus infection
Authors: Reznikov Elizabeth A , Hoeflinger Jennifer L , Monaco Marcia H , Miller Michael J , Donovan Sharon M
Published: June 2021

Staphylococcus aureus (S. aureus) is a primary cause of death due to sepsis in neonatal intensive care units. Fatal sepsis is characterized by systemic inflammatory response syndrome (SIRS), leading to systemic inflammation and organ dysfunction. Herein, a clinically‐relevant piglet model of SIRS secondary to S. aureus infection was developed. Colostrum‐deprived pigs had umbilical catheters placed within 12h of birth and were fed sow's milk replacer formula. On d7, piglets were either non‐infected (n=2) or injected IV with 1ml/kg BW S. aureus (strain S54F9) at 103(low; n=3) or 105(high; n=3) CFU/ml. Blood was repeatedly sampled from the catheter to follow the course of cytokine response and piglets were euthanized 10–14d post‐infection. Most infected animals developed a transient fever (>;103.5°F) post‐infection. Infected animals had elevated WBC count and percentage of neutrophils, characteristic of SIRS. S. aureus was detected in the spleen (n=5), kidney (n=3), lung (n=3) and heart (n=2) of infected animals and high‐dose pigs developed rear limb septic arthritis confirmed by culture. Serum cytokine responses were variable; however, IL‐10 was highest in the first 24 h post‐infection, whereas IL‐6 peaked between 72–96 h post‐infection. In conclusion, we have established a piglet model of systemic S. aureus infection that can be used to investigate therapeutic nutritional interventions.
The protective effects of Alginate and Fucoidan from the brown seaweed on the alcohol‐induced liver damage
Authors: Kim Taeseong , Kim Ye‐Jin , Lim Jung Dae , Sohn Eun‐Hwa
Published: June 2021

Alcohol‐induced liver injury progresses from fatty infiltration followed by a harmful cause of inflammation leading to an irreversible damage. In this study, two compounds (alginate and fucoidan) isolated from marine edible brown algae were examined for their protective effects against ethanol‐induced toxicity in vitro and in vivo. With response to ethanol‐challenge, fucoidan at 50 μg/mL was decreased TGF‐β1 expression, a hallmark of liver fibrosis and iNOS expression in HepG2 cells. The administration of 30 mg/kg of fucoidan resulted in the reduction of sGOT and sGPT levels caused by 25% ethanol exposure (2.0 g/kg/day, PO) for 5 days in mice and reduced TGF‐b1 expression in liver. These findings suggest that fucoidan may be a potential candidate for attenuated of ethanol induced liver damage. This work (Grants No. C0034917) was supported by Business for Cooperative R&D between Industry, Academy, and Research Institute funded Korea Small and Medium Business Administration in 2012.
The Effect of Omega‐3 Polyunsaturated Fatty Acid and Soy Protein Isolate Supplementation on Kidney Function in Female Polycystic Kidney Diseased Rats
Authors: Maditz Kaitlin Hope , Mock Kaitlin , Knecht Adam , Nanda Nainika , Oldaker Christopher , Tou Janet
Published: June 2021

Polycystic kidney disease (PKD) is a genetic disorder characterized by multiple cyst formation that increases renal size, structural damage, and loss of function. Studies showed soy protein isolate (SPI) protective against chronic kidney disease and omega‐3 polyunsaturated fatty acids (n‐3 PUFAs) decrease inflammation. Few studies have examined the role of diet in PKD. The study objective was to investigate whether SPI and/or n‐3 PUFA supplementation attenuates PKD progression. Young (age 28 d) female pck rats were randomly assigned (n=12/group) to diets consisting of casein + corn oil (Casein + CO), casein + soybean oil (casein + SO), SPI + soybean oil (SPI + SO) or SPI + 1:1 soybean/salmon oil (SPI + BLEND) for 12‐weeks. Kidney weights were highest in the SPI + BLEND group. Histology showed interstitial inflammation, fibrotic changes, interstitial matrix deposition, and structural effacement due to cyst growth in all groups. Cortical cyst obstruction indicating more extensive nephron damage was only observed in the SPI + BLEND group. The SPI +BLEND group had the highest (11.78 + 0.60 mg/dL, P=0.01) and Casein + SO group the lowest (9.28 + 0.81, P=0.01) serum blood urea nitrogen. Urinary calcium and phosphorus was higher in the casein than SPI groups. There were no significant differences in serum calcium, phosphorus, uric acid, total protein or albumin. Based on the results, diet did not attenuate PKD progression.

Grant Funding Source: WVU Hatch Grant H459
Cardiomyocytes hypertrophy in vitro are induced by TLR2 and TLR4 agonists through NF‐kB pathway
Authors: Silva Fernanda Gaisler , Ramos Marcela Sorelli Carneiro
Published: June 2021

Introduction

Toll Like receptors, a class of specific proteins involved in innate immune activation, exert innumerous effects in a vast number of tissues and organs, including cardiac trophism modulation. The objective of this study is to evaluate TLR2 and TLR4 agonists effect in hypertrophy cardiomyocytes in vitro and the role of NF‐kB pathway.

Methods

Primary cardiomyocytes cultures were prepared from neonatal Wistar rats by collagenase/pancreatin method. Cardiomyocytes were treated for 24h with LPS (TLR4 agonist) and Zymozan (TLR2 agonist). Gene expression analyzes: ANF, Alpha‐actin, TLR2, TLR4, MyD88 and NF‐kB were evaluated by Real‐time PCR. Protein expression of TLR2, TLR4, MyD88 and NF‐kB were evaluated by Western Blotting. Results were expressed as mean±SD and p<0.05 and p<0.01.

Results

LPS and Zym promoted an increase on ANF and alpha‐actin expression, both cardiomyocytes hipertrophy markers. The MyD88 mRNA levels increased after both LPS and Zym treatments and protein levels increased after Zym treatment. NF‐kB mRNA and protein levels increased after both treatments. These results suggest an important role for TLRs and NF‐kB pathway in cardiomyocyte hypertrophy in vitro.

Financial support: FAPESP.
Reduced BDNF attenuates inflammation and angiogenesis to improve survival and cardiac function following myocardial infarction in mice
Authors: Halade Ganesh V , Ma Yonggang , Ramirez Trevi R. , Zhang Jianhua , Dai Qiuxia , Hensler Julie G. , Lopez Elizabeth F , Ghasemi Omid , Jin Yu‐Fang , Lindsey Merry L.
Published: June 2021

Brain‐derived neurotrophic factor (BDNF) increases in failing hearts, but BDNF roles in cardiac remodeling following myocardial infarction (MI) are unclear. Male BDNF+/+ (wild type, WT) and BDNF+/− heterozygous (HET) mice at 6–9 months of age were subjected to MI and evaluated at day (d) 1, 3, 5, 7, or 28 post‐ MI. At d28 post‐MI, 76% of HET vs. 40% of WT mice survived, while fractional shortening was improved and neovascularization levels were reduced in the HET (all p<0.05). At d1 post‐MI, matrix metalloproteinase (MMP)‐9 and myeloperoxidase (MPO) increased in WT, but not in HET. Concomitantly, monocyte chemotactic protein (MCP)‐1 and ‐5 levels increased and vascular endothelial growth factor (VEGF)‐A decreased in HET. Neutrophil infiltration peaked at d1–3 in WT mice, and this increase was blunted in HET. To determine if MPO administration could rescue the HET phenotype, MPO was injected at 3h post‐MI. MPO restored VEGF‐A levels without altering MMP‐9 or neutrophil content. In conclusion, reduced BDNF levels modulated the early inflammatory and neovascularization responses, leading to improved survival and reduced cardiac remodeling at d28 post‐MI. Thus, reduced BDNF attenuates early inflammation following MI by modulating MPO and angiogenic response through VEGF‐A.

Funding support: GVH: NIH‐1K99AT006704–01, YFJ: NIH EB009496 and 1SC2 HL101430, MLL: NIH/NHLBI HHSN 268201000036C (N01‐HV‐00244), R01 HL075360, the Max and Minnie Tomerlin Voelcker Fund, and the Veteran's Administration.
Influence of late exercise training on myostatin and follistatin expression in soleus muscle of rats with chronic heart failure
Authors: Guizoni Daniele Mendes , Junior Silvio Assis Oliveira , Martinez Paula Felippe , Lima Aline Regina Ruiz , Bonomo Camila , Damatto Ricardo Luiz , Cezar Marcelo Diarcadia Mariano , Pagan Luana Urbano , Zornoff Leonardo Antonio Mamede , Okoshi Katashi , Okoshi Marina Politi
Published: June 2021

In this study we evaluated effects of late exercise training on myostatin and follistatin expression in soleus muscle of rats with myocardial infarction (MI)‐induced heart failure. Three months after coronary occlusion, rats were assigned to four groups: Sham sedentary (Sh‐Sd, n=12), Sham exercised (Sh‐Ex, n=12), MI sedentary (MI‐Sd, n=11), and MI exercised (MI‐Ex, n=10). Physical training was performed on treadmill, 40 min/day, 5 days/week, 16 m/min for 12 weeks. Left ventricular (LV) function was evaluated by echocardiogram. Infarct size and soleus fiber cross‐sectional area were evaluated in histological sections. Myostatin and follistatin protein levels were assessed by Western blot.

Statistical analyses

ANOVA and Bonferroni. MI size did not differ between MI groups (p>;0.05; t test). LV fractional area change was higher in MI‐Ex than MI‐Sd (Sh‐Sd 70.1±6.4; Sh‐Ex 70.0±5.7; MI‐Sd 22.8±7.9; MI‐Ex 29.7±5.5%). Myostatin levels were lower in MI‐Sd than both Sh‐Sd and MI‐Ex (Sh‐Sd 1.00±0.35; Sh‐Ex 1.02±0.37; MI‐Sd 0.66±0.18; IM‐Ex 1.17±0.48 arbitrary units). Follistatin levels and cross‐sectional area did not differ between groups. In conclusion, late exercise training increases frequency of skeletal fibers splitting and prevents myostatin expression change in soleus muscle of infarcted rats. Research support: FAPESP.
Mechanism underlying the antioxidant activity of taurine
Authors: Jong Chian Ju , Schaffer Stephen
Published: June 2021

Taurine is an ubiquitous sulfur‐containing amino acid found in high concentration in the heart. Although it has antioxidant activity, it is not a classical free radical scavenger, therefore, its mechanism remains unclear. Recently we proposed that taurine prevents the formation of superoxide by the mitochondria, a mechanism related to the formation of a post‐transcriptional modification of tRNALeu(UUR) in the mitochondria. By decreasing its post‐transcriptional wobble modification, taurine deficiency reduces the levels of respiratory chain subunits, thereby diminishing respiratory chain complex activity and oxygen consumption. As predicted, taurine‐depleted hearts are oxidativestressed, as exemplified by decreased aconitase activity and the glutathione redox ratio. Among other damage, taurine deficiency triggers cell death, as shown by an increase in the percentage of apoptotic cells. Because the antioxidant, Tiron, prevents apoptosis in taurine deficient cardiomyocytes in culture, cell death is attributable to oxidative stress. In conclusion, this study confirms the potential role of taurine as an antioxidant, but not as a radical scavenger. Rather, taurine ensures an efficient flux of reducing equivalents through the respiratory chain, thereby preventing the diversion of electrons to the acceptor oxygen, resulting in the formation of reactive oxygen species. This work was supported by AHA grant.
Cisplatin nephrotoxicity involves mitochondrial injury with impaired tubular mitochondrial enzyme activity
Authors: Zsengeller Zsuzsanna Kinga , Mukhopadhyay Partha , Kalyanaraman Balaraman , Parikh Samir , Karumanchi Ananth S. , Stillman Isaac E. , Pacher Pál
Published: June 2021

Cisplatin is a commonly used chemotherapeutic drug, the clinical use of which is limited by the development of dose‐dependent nephrotoxicity. Recent studies have suggested that mitochondrial dysfunction in tubular epithelium contributes to cisplatin‐induced nephrotoxicity (PMID: 22120494). Here we extend those findings by describing the role of an important electron transport chain enzyme, cytochrome c oxidase (COX). Immunohistochemistry for COX 1 protein demonstrated that, in response to cisplatin, expression was mostly maintained in focally damaged tubular epithelium. In contrast, COX enzyme activity in proximal tubules (by light microscopy) was decreased. Ultrastructural analysis of the cortex and outer stripe of the outer medulla (OSOM) showed decreased mitochondrial mass, disruption of cristae and extensive mitochondrial swelling in proximal tubular epithelium. Functional electron microscopy (EM) showed that COX enzyme activity was decreased in the remaining mitochondria in the proximal tubules, but maintained in distal tubules. A single systemic dose of mitochondrially targeted anti‐oxidant prevented mitochondrial injury and dysfunction. In summary, cisplatin induced nephrotoxicity is associated with structural and functional damage to the mitochondria. Mitochondrially targeted antioxidants may offer a novel therapeutic approach to prevent cisplatin nephrotoxicity.
Induction of Apoptosis in the Bone Marrow Promotes Regenerative Actions of Parathyroid Hormone (PTH) in Bone.
Authors: Koh Amy J , Cho Sun W , Entezami Payam , Pettway Glenda , McCauley Laurie
Published: June 2021

PTH has bone regenerative properties via mechanisms that remain elusive. This study aimed to determine the impact of marrow apoptosis on PTH anabolic actions. Studies were in conformance with FASEB animal use principles. Adult mice were treated with pro‐apoptotic etoposide (ETOP) or vehicle (VEH), daily 5d then 3d/wk for 6wks. PTH or VEH was administered daily for: VEH, PTH, ETOP and PTH+ETOP and skeletal phenotyping, flow cytometry, and serum bone markers analyzed. MicroCT and histomorphometry of tibiae revealed ETOP decreased and PTH increased bone volume and area, the greatest increase with PTH + ETOP. ETOP (5d) increased early marrow apoptotic cells (AnnexinV+PI‐) as well as MER+CD11b+F4/80+ and MER+CD68+F4/80+ myeloid cells. At 6wks, CD68+, Gr1+CD11b+F4/80+, CD45+and Lin−CD29+sca1+ cells were increased in ETOP w/ or w/o PTH. Serum TRAcP5b was increased in ETOP and to a greater extent with PTH. P1NP levels were increased in PTH and PTH+ETOP. PTH+ETOP mice had reduced osteoclasts per tibial bone vs. VEH or ETOP. In another model, luciferase+bone marrow stromal cells were implanted to form ectopic ossicles and bone growth tracked via bioluminescence. ETOP mice had increased luciferase vs. VEH, and ETOP augmented the PTH increase. These data suggest that induction of apoptosis provides a conducive environment for PTH pro‐regenerative actions, effects which may be due to increased macrophage efferocytosis.
Caspase‐generated neoepitopes as markers of axonal degeneration in neural development and injury
Authors: Sokolowski Jennifer D , Heffron Dan S , Deppmann Christopher , LeBlanc Andrea C , Mandell James W
Published: June 2021

Neuronal death, axonal degeneration, and synapse loss are hallmarks of central nervous system development and injury. Commonly employed markers of apoptosis are TUNEL and immunohistochemistry for activated caspases, but these often fail to report axonal degeneration associated with somatic apoptosis. Axonal degeneration is classically detected by silver degeneration stains, but these have limitations (non‐selectivity, minimal reproducibility and incompatibility with co‐labeling). We demonstrate that antibodies to caspase‐cleaved protein substrates beta‐actin (cleaved by caspase 3 at D244) and alpha‐tubulin (cleaved by caspase 6 at D438) enable superior visualization of neurite degeneration. These antibodies label apoptotic cell bodies and neurites during embryonic and postnatal developmental pruning. In addition, they reveal injured neurons and neurites after induction of apoptosis in a mouse model of fetal alcohol syndrome and in sympathetic neuron cultures induced to undergo apoptosis after nerve growth factor withdrawal. Finally, the markers may have clinical utility, as they highlight degenerating neurites in neonatal hypoxic/ischemic injury and in adult patients with multiple sclerosis. Ongoing work aims to identify novel compartment‐specific neoepitopes that will allow monitoring of somatodendritic, axonal, and synaptic remodeling and degeneration. Supported by NS075869.
Epigenetic Regulation of PTEN in Hepatic Cancer
Authors: Qadir Ximena Vanegas , Han Chang , Wu Tong
Published: June 2021

The pathogenesis of hepatocellular carcinoma (HCC) remains misunderstood offering limited therapeutic options for patients with hepatic cancer.

Treatment of HCC cell line (Huh‐7) with DNMT inhibitor, Decitabine, and HDAC inhibitor, Trichostatin A, increased the expression of PTEN protein. Methylation‐specific qPCR assay showed Decitabine/TSA treatment resulted in hypermethylation of CpG islands located on the PTEN gene promoter. Furthermore, miR‐185 targets DNMT1, thus upregulating the expression of PTEN in Huh‐7. Forced overexpression of miR‐185 abolished the expression of DNMT1 protein and enhanced the level of PTEN protein in Huh‐7 stable cells. MiR‐185 xenograft tumor volume from SCID mice was significantly decreased versus control. Likewise, Ki‐67 immunostaining in Huh‐7 control stable cell line showed a higher proliferative. Xenograft protein samples showed increased PTEN expression and decreased DNMT1 activity. DNMT‐1 3'UTR reporter plasmids were constructed/cloned showing the wild type‐3'UTR of DNMT1 was significantly depleted in Huh‐7 miR‐185 stable cell line versus control in luciferase assay experiments. Co‐transfection of Huh‐7 miR‐185 ORF versus control showed DNMT1 restoration via western blot, Invasion and Cell Proliferation Assay. In‐situ hybridization showed a substantial decrease of staining expression in human HCC tumor samples versus normal liver tissue probed for miR‐185.

Our results demonstrate a novel epigenetic regulation of PTEN by miR‐185, which is critical in hepatic carcinogenesis.