bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Folic Acid Deficiency on the Murine Mammary Gland
Authors: Miszewski Susan , Berryhill Grace , Green Ralph , Borowsky Alexander , Miller Joshua , Hovey Russell
Published: June 2021

The epithelial ducts within the mammary glands (MG) undergo phases of isometric and allometric growth during early, postnatal life. Breast cancer is suggested to have origins in early life, a period during which folate requirements are high. We investigated the effects of folic acid (FA) on early MG development. Dams were assigned to either a control (2 mg FA/kg), deficient (0 mg FA/kg), excess (5 mg FA/kg), or super excess (20 mg FA/kg) amino acid defined diet and offspring continued to receive the same diet as their respective dams through postnatal and pubertal development. MG tissues from offspring were assessed at weaning and in early puberty. We observed no significant effects of excess or super excess FA diets on the MG. The area occupied by the mammary ducts at weaning and during early puberty was reduced in FA deficient animals compared with controls (p<0.0001). The rate of MG growth in the isometric phase, from birth to weaning, was less for the MG of deficient females compared with those from controls (slope 95% confidence limits 0.23‐0.78 versus 1.47‐1.82, respectively). The subsequent rate of allometric growth was unaffected by diet. The supramammary lymph nodes in deficient animals were smaller than in control animals alongside a reduction in the area of the thymic medulla. Epigenetic and local expression analysis of signaling and growth factors in the MG are ongoing. We conclude that dietary FA deficiency alters normal development of the epithelial ducts and lymph node in the MG of prepubertal female mice. Funding source: CA Breast Cancer Research Program Grant #18IB‐000
Dysfunctional Cardiac Fatty Acid Metabolism in Cystathionine‐beta‐Synthase +/‐ Mice with Obesity
Authors: Glier Melissa , Wambolt Rich , da Silva Robin , Gerrard Sarah , Aleliunas Rika , Jacobs Rene , Allard Michael , Devlin Angela
Published: June 2021

Obesity‐related cardiac lipid accumulation is associated with toxicity and dysfunction. Cysteine is required for the synthesis of the antioxidant glutathione, supplied by the transsulfuration of homocysteine by cystathionine‐beta‐synthase (Cbs). Cbs+/‐ mice with diet‐induced obesity have reduced cardiac glutathione, and greater glucose intolerance and cardiac lipotoxicity. Our objective was to determine the functional effects and mechanisms of cardiac lipotoxicity in Cbs+/‐ mice with diet‐induced obesity. Cbs+/‐ and Cbs+/+ mice were fed a control‐diet (CD) or a high‐fat diet (HFD) from weaning for 20 weeks. Cbs+/‐ mice fed CD had less visceral adiposity compared to Cbs+/+ mice. Ex vivo working heart assessments revealed greater palmitate oxidation in mice fed the HFD than CD. Independent of diet, Cbs+/‐ mice had reduced palmitate oxidation compared to Cbs+/+ mice. Cbs+/‐ mice had larger hearts and more arachidonic acid (AA) and docosahexaenoic acid (DHA) concentrations in the heart than Cbs+/+ mice. Mice fed the HFD had higher AA, but lower DHA concentrations in the heart than mice fed the CD; effects greatest in Cbs+/‐ mice. A lower ratio of phosphorylated‐AMPKα/AMPKα expression was observed in Cbs+/‐ than Cbs+/+ mice, and this occurred to a greater extent in those fed the HFD. Cbs+/‐ mice with diet‐induced obesity have altered cardiac fatty acid metabolism and larger hearts, further supporting a role for Cbs in obesity‐related cardiac lipotoxicity.

Supported by NSERC and CIHR
Maternal intake of vitamin B6 and maternal and cord plasma levels of pyridoxal 5' phosphate in a cohort of Canadian pregnant women and newborn infants
Authors: Kim Denise , Plumptre Lesley , Masih Shannon , Visentin Carly , Sohn Kyoung‐Jin , Ly Anna , Lausman Andrea , Berger Howard , Croxford Ruth , O'Connor Deborah , Kim Young‐In
Published: June 2021

Vitamin B6 is important in fetal development as an essential coenzyme involved in 1‐carbon metabolism. However, maternal intake and blood levels of B6 during pregnancy have not been well established. We determined dietary and supplemental B6 intakes from 368 Canadian pregnant women using a FFQ and measured pyridoxal 5' phosphate (PLP) levels in their plasma at baseline (11‐16 wks gestation), delivery and in umbilical cord plasma. Mean dietary B6 intake(95% CI) at baseline and delivery were 1.6(1.5‐1.7) and 1.6(1.5‐1.6) mg, respectively. 88% reported B6 supplement use during pregnancy; 75% met the RDA (1.9 mg) for B6 through supplement use. Geometric mean (95%CI) PLP levels were 110(101‐119), 61(56‐65), and 297(277‐319) nmol/L at baseline, delivery and in cord blood, respectively (p<0.0001). 11 and 24% of pregnant women had suboptimal PLP levels (<20 nmol/L) at baseline and delivery, respectively. 58% of the women with suboptimal PLP had B6 intake of at least the RDA. Maternal PLP levels from supplement users were higher than those from nonusers at baseline (p=0.001) but not at delivery (p=0.47). Cord PLP levels from users were higher than those from nonusers (p=0.005). Although 75% of the women met the RDA for B6 mostly through supplements, 11‐24% had suboptimal plasma PLP levels. B6 supplementation may not be sufficient for adequate maternal PLP levels during pregnancy. Future studies are warranted to determine optimal B6 intake and blood levels during pregnancy. Funded by: CIHR (MOP#106446)
A Decrease in γ‐Tocopherol Associates with Impaired Muscular Performance After Anterior Cruciate Ligament Surgery
Authors: Barker Tyler , Henriksen Vanessa , Rogers Victoria , Trawick Roy
Published: June 2021

We tested the hypothesis that a decrease in γ‐tocopherol (γT) increases serum cytokine concentrations and impairs muscular performance after an anterior cruciate ligament (ACL) injury and surgery. Subjects were randomly assigned to one of three groups: (1) placebo (n=14), (2) multi‐vitamin/‐mineral (MV; n=14), or (3) vitamins E (α‐ [600 mg RRR‐α‐tocopherol, αT)] and γ‐ [600 mg of RRR‐γT]) and C (1000 mg ascorbic acid, AA) (EC; n=15). Supplements were taken daily starting ~2‐wk prior to and concluding 16‐wk after surgery. Fasting blood samples were obtained and single‐leg peak isometric force measurements were performed at baseline (Bsl, prior to supplementation), before surgery (~120‐min – blood draw only), and 8‐wk, 12‐wk, and 16‐wk after surgery. An exercise protocol consisting of repetitive knee and hip extension and flexion contractions to exhaustion was performed on the injured limb at 16‐wk. αT, γT, AA, and cytokines were measured in each blood sample, and peak isometric force was measured on injured and non‐injured legs separately at each testing session. Plasma αT and AA increased (∼42% and 28%, respectively) in the MV and EC groups, while γT decreased (∼47%) in the MV and increased (∼153%) in the EC group. Serum cytokines and peak isometric forces were not different between groups. Time to exhaustion during the exercise protocol decreased (∼30%) in the MV group and correlated with the change (Bsl to 16‐wk) in plasma γT in the MV and EC groups (n=29, r=0.42, p<0.05). We conclude that a decrease in circulating γT hinders muscular performance after ACL reconstruction.
Stable iron isotopic techniques and plasma hepcidin profiles to optimize iron supplementation regimens
Authors: Moretti Diego , Goede Jeroen , Christophe Zeder , Swinkels Dorine , Tjalsma Harold , Melse Alida , Zimmermann Michael
Published: June 2021

Background

Oral iron supplementation is used to treat iron deficiency, but absorption is often low and correction is variable.

Objective

Determine the duration and magnitude of the plasma hepcidin (pHep) response induced by oral iron (Fe) supplements and concomitantly measure bioavailability in healthy iron depleted young women.

Methods

pHep (measured by C‐ELISA), iron status and inflammation were monitored at regular intervals. On day 1, no supplements were given (control day). On days 2 and 3, subjects received iron supplements containing 40, 60, 80, 160 or 240 mg Fe as FeSO4 as either single or two consecutive daily doses extrinsically labeled with stable iron isotopes. Iron bioavailability was measured by the isotopic enrichment of erythrocytic iron 14 days after administration.

Results

Both Fe dose (P<0.05) and time of day (P<0.05) were associated with increase in pHep. Compared to control days, pHep was significantly higher at 8h and 24h after administration for 60, 80, 160 and 240 mg (P<0.05) but not for 40 mg Fe. Total Fe absorption from the Fe dose on the second day of administration compared to a single Fe dose on the first day was decreased for all dosages above 40 mg( P<0.01). With twice per day dosing (60 mg Fe) the afternoon dose was less bioavailable (P<0.05).

Conclusion

In Fe‐depleted women, consecutive day doses of supplemental Fe at 60 mg or above increase pHep and decrease fractional Fe bioavailability. These new data will help guide optimal dosing regimens.
Chromium intake and its relationship with blood glucose in healthy Korean adults
Authors: Choi Mi‐Kyeong , Kim Mi‐Hyunn , Bae Yun‐Jung , Lee Haeng‐Shin
Published: June 2021

Chromium (Cr) is the most abundant element in the Earth's crust and widely distributed throughout the food supply. Previous research has demonstrated the beneficial effects of Cr on circulating glucose and insulin; however, the dietary intake level of Cr in the self‐selected diet and its relationship with blood glucose have not been investigated because of the lack of a database on Cr content in commonly consumed foods. Thus, the purpose of this study was to assess Cr intake and investigate its relationship with blood glucose in healthy adults. A total of 51 adults aged 20‐28 years participated in this study and their Cr intakes were estimated by using the recently established database. Fasting blood glucose levels of the subjects were analyzed as well. Daily energy intakes of men and women were 1733.41 kcal and 1571.42 kcal, respectively, and daily Cr intakes were 66.38 ug and 52.34 ug, respectively, without significant difference. The subjects were supplied Cr mainly from cereals (20.04 ug/day), followed by vegetables (8.36 ug/day), beverages (6.00 ug/day), and fish and shellfishes (5.24 ug/day). Fasting blood glucose showed significant correlations with Cr intake from cereals in men (r = ‐0.4995, p < 0.05), total Cr intake in men (r = ‐0.4366, p < 0.05), and Cr intake from fish and shellfishes in women (r = 0.4248, p < 0.05), respectively. These results show that higher Cr intake from diet may be associated with lower blood glucose levels in healthy Korean adults.
Egg Yolks Attain Anti‐inflammatory Properties after Partial Delipidation
Authors: Shen Qiwen , Riedl Kenneth , Cole Rachel , Lehman Christopher , Xu Lu , Alder Hansjuerg , Belury Martha , Schwartz Steven , Ziouzenkova Ouliana
Published: June 2021

Egg yolk (EY) contains bioactive lipophilic compounds with potential beneficial and deleterious inflammatory and metabolic properties. How the composition of EY influences activation of NF‐kB, a key transcription pathway in inflammation, remains unclear. We hypothesized that a partial delipidation of EY, without changes in EY's carotenoids and vitamin content, was sufficient to inhibit NF‐kB activation. We performed a hexane extraction of EY that removed 20‐30% of cholesterol and triglycerides. The remaining fraction was termed EY‐Polar fraction. NF‐kB activation in response to EY fractions was examined in 3T3‐L1 adipocytes using a NF‐kB response element reporter assay and by analyzing expression of 248 inflammatory genes. Adipocytes stimulated with EY and EY‐Non‐polar fractions have similar levels of NF‐kB activation. Although EY‐Polar and EY fractions contained similar level of vitamins, carotenoids, and fatty acidsonly delipidated EY‐Polar fraction significantly suppressed NF‐kB activation via down‐regulation of toll like receptor‐2 and up‐regulation of inhibitory Tollip and Ly96. SREBP inhibitor fatostatin prevented NF‐kB activation in the presence of EY‐Non‐polar fraction, suggesting that crosstalk between these pathways. Our data suggest that non‐polar lipids in EY sensitize NF‐kB in adipocytes in vitro likely via the SREBP pathway. Thus, moderate delipidation of the yolk of regular eggs from different suppliers may improve their beneficial properties.
Dried Plums Promote Increased Antioxidant Capacity in Smokers and Nonsmokers
Authors: Zawilski Alexandra , Nelson Stephanie , McGill Brittany , McIntosh Marian , Hong Mee Young , Hooshmand Shirin , Kern Mark
Published: June 2021

Smokers suffer from decreased antioxidant capacity, a risk factor for numerous chronic diseases. Consumption of antioxidant‐rich foods such as dried plums may increase antioxidant capacity and reduce disease risk. Nonsmokers (n=14) and smokers (n=6) between the ages of 18 and 45 years were recruited to participate in 2 randomly ordered trials in which they received 100 g of dried plums at one visit and an isocaloric control food (muffins) at another. Antioxidant capacity was examined in blood samples collected at baseline and 60, 90, and 120 minutes post‐feeding. No significant differences in postprandial antioxidant capacity were detected over time when consuming the muffin snack in smokers or nonsmokers. Dried plum consumption elicited significant increases (p<.05) in antioxidant capacity in both groups together from baseline throughout all time points, with peak values achieved at 90 minutes. Dried plums promoted higher antioxidant capacity compared to muffins at all time points (p<.05). Smokers had significantly lower (p<.05) levels of antioxidant capacity at all time points compared to nonsmokers. Although antioxidant capacity remained elevated (p<0.05) at each time point in comparison to baseline, it decreased (p<0.05) from minutes 90 to 120 for smokers but continued to rise up to the 120 min time point for nonsmokers. Results suggest that dried plums are efficient in increasing antioxidant capacity for both smokers and nonsmokers and that they may be an effective way to reduce disease risk factors in smokers that are struggling with cessation. Future research should evaluate the use of dried plums as a chronic intervention to promote health of smokers. Funded by the California Dried Plum Board.
Evaluation of analytical methods for measuring plasma antioxidant capacity by comparing with urinary 8‐isoprostane concentration
Authors: Lee Sang Gil , Wang Taoran , Vance Terrence , Hurbert Patrice , Koo Sung I. , Chun Ock K.
Published: June 2021

Although several analytical methods for measuring the total antioxidant capacities (TAC) of foods have been applied to biological samples, different methods often yield dissimilar results due to the types of radical used (ABTS or DPPH radical) and the principles of assays involved.Thus, this study aimed to evaluate the validity of three commonly used methods of measuring plasma TAC: the ABTS, DPPH, and ferric reducing antioxidant power (FRAP) assays against urinary 8‐isoprostane as an established biomarker of oxidative stress. The TAC of ninety‐six human plasma samples were measured using each of the three methods and compared with urinary 8‐isoprostane concentrations adjusted to urinary creatinine. Among the three methods, only the ABTS and FRAP assays were correlated with each other (r = 0.58, p < 0.0001). The agreement between each of the 3 methods and 8‐isoprostane was determined using the prediction intervals of linear regression expressed as mean of 8‐isoprostane ± 95% of prediction interval. The means of TAC by the ABTS, DPPH, and FRAP assays corresponding to 8‐isoprostane levels were 6.98 ± 7.54 for ABTS, 6.98 ± 7.78 for DPPH and 7.09 ± 7.85 ng/mmol creatinine for FRAP assays. The ABTS method better agreed with 8‐isoprostane than the other methods with lower 95% of prediction interval. Furthermore, plasma TAC determined by the ABTS assay were significantly correlated with urinary 8‐isoprostane levels (r = ‐0.21, p = 0.04). The results indicate that the ABTS assay is a reliable method for measuring plasma antioxidant capacity and predicting the body's oxidative stress status. This study was supported by the Nutricia Research Foundation's Research Grant.
Dietary Strawberries Increase Glutathione in Obese Participants with Above Optimal Serum Lipids
Authors: MORRIS STACY , BETTS NANCY , LYONS TIMOTHY , BASU Arpita
Published: June 2021

Background

Dietary flavonoid intake, especially berry flavonoids, has been associated with antioxidant functions and metal chelation in mechanistic studies. Purpose: We examined the hypothesis that freeze‐dried strawberries (FDS) improve antioxidant biomarkers in adults with abdominal adiposity and elevated serum lipids. Methods: In a randomized dose‐response controlled trial, 60 volunteers [5 men and 55 women; age: 49 ± 10 years; BMI: 36 ± 5 kg/m2 (mean±SD)] were assigned to one of the following arms: low dose FDS (LD‐FDS) (25g/d), low dose control (LD‐C), high dose FDS (HD‐FDS) (50g/d), and high dose control (HD‐C) beverages for 12 weeks. Control beverages were matched for calories and total fiber. Serum levels of trace elements (copper, iron, selenium, zinc) and whole blood glutathione (GSH), and catalase activity were examined at screening (0 week) and after 12 weeks intervention. Results: At 12 weeks, glutathione levels were higher in HD‐FDS vs. LD‐FDS, as well as vs. HD‐C, and catalase activity was lower in HD‐FDS vs. baseline (all P<0.05). No differences were noted in serum trace elements following FDS intervention.

Conclusions

Dietary strawberries, especially at high doses may selectively modulate antioxidant biomarkers that influence risk factors of chronic diseases. These results need further confirmation in larger trials.
Extruded Amaranth (Amaranthus hypochondriacus) Hydrolysates Showed Potential Anti‐atherosclerotic Effect on THP‐1 Human Cells
Authors: Milán‐Noris Ada , Montoya‐Rodríguez Alvaro , Milán‐Carrillo Jorge , Dia Vermont , Cuevas‐Rodríguez Edith , Reyes‐Moreno Cuauhtémoc , González de Mejía Elvira
Published: June 2021

The aim of this study was to compare the anti‐atherosclerotic effect of unprocessed amaranth hydrolysate (UAH) and extruded amaranth hydrolysate (EAH) in lipopolysaccharide (LPS)‐induced human THP‐1 cells by protein expression, immunocytochemistry and confocal microscopy. Amaranth was extruded in a single screw extruder at 125 °C and 130 rpm. Unprocessed and extruded amaranth flours were hydrolyzed with pepsin and pancreatin and freeze dried until use. THP‐1 human cells, derived from acute monocytic leukemia, were treated with both hydrolysates at 1 mg/mL for 24 h; LPS was applied to induce inflammation. UAH and EAH inhibited the expression of interleukins related with the atherosclerotic process such as interleukin‐4, interleukin‐6, among others. Likewise, UAH and EAH inhibited the expression and production of MCP‐1 and tumor necrosis factor‐α (TNF‐α), respectively. Western blot analysis showed that EAH inhibited LOX‐1 (27%), ICAM‐1 (28%) and MMP‐9 (19%) expression, important protein markers in atherosclerosis. By confocal microscopy, EAH led to a reduction of 58, 52 and 79% for LOX‐1, ICAM‐1 and MMP‐9, respectively. In conclusion, extruded amaranth hydrolysates showed potential anti‐atherosclerotic in LPS‐induced THP‐1 human macrophage‐like cells, reducing expression of proteins associated with LOX‐1 signaling, perhaps attributed to the formation of bioactive peptides during the extrusion process
Effect of Grape Consumption on Biomarkers of Endothelial and Cardiovascular Health
Authors: Hu Qian , Randolph Jody , Giebler Kathryn , Stevens Allison , Steinberg Francene
Published: June 2021

Polyphenols have been associated with cardio‐protective benefits, however causal mechanisms remain elusive. This randomized, double‐blind, crossover study examines acute and longer‐term effects of grape consumption on biomarkers of endothelial and cardiovascular health. It was hypothesized that polyphenol‐rich grape consumption would protect against diet‐induced elevations and long‐term impairment in biomarkers of endothelial dysfunction during the acute postprandial period and after 4 weeks respectively. Twenty obese (BMI 37.0±9.9) but otherwise healthy adults were provided with isocaloric sachets of freeze‐dried whole grape powder (grape) (1 60g sachet/day=297mg polyphenol) and placebo (1 60g sachet/day=0mg polyphenol) for daily consumption, each treatment for 4 weeks separated by 2 week washout. To examine acute effects, postprandial blood samples were collected at 0, 1, 3 and 5 hours following a high fat high carbohydrate (HFHC) meal challenge and grape or placebo consumption. Longer‐term effects were examined by fasted blood samples collected on the last day of each 4‐week intervention. HDL increased (p<0.05) after 4 week grape consumption, but no other significant changes in lipids or blood glucose were noted between groups. Data analyzed by repeated measures, 2‐way ANOVA, showed that PAI‐1 and MCP‐1 were significantly elevated in plasma after HFHC challenge (p<0.05). However, no significant differences in plasma sICAM, sVCAM, ET‐1, PAI‐1 or MCP‐1 were detected between grape and placebo treatments after HFHC challenge or 4‐week intervention. Grape consumption has some beneficial effects on markers of cardiovascular health.

Supported by California Table Grape Commission and USDA CRIS CA‐S*‐NTR‐6316‐H
The Change in Human Microvascular Function and its Relationship to Plasma Epoxide Content After Short‐Term Walnut Intake
Authors: Holt Roberta , Yim Sun , Shearer Gregory , Keen Carl , Djurica Dragana , Newman John , Hackman Robert
Published: June 2021

Walnuts are a source of alpha‐linolenic acid (ALA) and linoleic acid (LA). We examined the effects of short‐term walnut intake on the reactive hyperemia index (RHI), a measure of microvascular function. We hypothesized that 4 weeks of 40g/day of walnuts would improve RHI compared to 5g/day of walnuts in hypercholesterolemic postmenopausal women. Microvascular function is influenced, in part, by locally produced vasodilators, such as nitric oxide and endothelial‐derived hyperpolarizing factor (EDHF). Putative EDHF include cytochrome P450 epoxides of arachidonic acid (AA) known as the EpETrEs. Epoxides are also derived from ALA (EpODE) and LA (EpODE); however, their vascular influence is currently undefined. Four weeks of 40g/day of walnut intake increased RHI compared to 5g/day (2.61 ± 0.10 versus 2.24 ± 0.12, respectively, p = 0.027). Plasma EpODE and EpOME were significantly increased with 40g/day compared to 5g/day of walnut intake. The total change in plasma epoxides was strongly associated with the change in RHI (r=0.65, p = 0.002). There was a strong association between the change in EpODE, EpOME, and EpETrE, as well as, these epoxides and the change in RHI (|r|=0.56, p < 0.01); with the strongest association observed for 14(15)‐EpETrE (r=0.72, p < 0.001). These data are consistent with reports of improved vascular function with walnut consumption, and provide new insights into potential relationships between changes in microvascular function and plasma epoxides after walnut intake.
Phenol Loss in Heating of Extra Virgin Olive Oil is Dependent on Temperature, Time, and Cooking Conditions
Authors: Flynn Mary , Wang Selina
Published: June 2021

Objective

The currently available studies that assess the effect of heat on extra virgin olive oil utilize cooking temperatures and times that would exceed normal home‐cooking conditions. We tested the effect of temperatures and cooking conditions reflective of home‐cooking on the content of total phenols, and individual phenols (e.g. oleocanthal, α‐tocopherol and hydroxytyrosol) in olive oils with a range of total phenol content.

Methods: olive oil with low (140 mg/kg), medium (300 mg/kg), and high (450 mg/kg) total phenol content was heated to 121°C and 180 °C on a stovetop for 10 minutes and baked in an oven for 30 minutes at 204 °C. After cooling, the oils were analyzed and some of the results are summarized in the table below.

10 mins @121°C (stove top)
10 mins @180°C (stove top)
30 mins @ 204°C (oven)

Phenol content (mg/kg)
Total phenol loss (%)
Oleocanthal loss (%)
Total phenol loss (%)
Oleocanthal loss (%)
Total phenol loss (%)
Oleocanthal loss (%)

Low (140)
2
21
58
53
27
38

Med. (300)
11
3
71
14
44
9

High (450)
41
18
79
35
44
32

Conclusions

Loss of total phenol content in extra virgin olive oil is lower for stove top cooking at common home cooking temperatures (121°C) and for baking (204°C) compared to frying temperature (180°C). Air exposure during the frying process may be responsible for the higher losses when compared with baking. Oleocanthal, an anti‐inflammatory compound, is relatively heat resistant. Future experiments should test the effects of heating the oils with food.
A traditional Mexican diet decreased oxidative stress, improves carbohydrate and lipid metabolism and modifies the microbiota in a diet‐induced obesity
Authors: Avila‐Nava Azalia , Noriega Lilia , Tovar Armando , Torres Nimbe
Published: June 2021

Corn, beans, chia and pumpkin seeds, nopal and tomato were part of the ancient Mexican diet with high antioxidant activity. Nowadays in Mexico there is decrease in the consumption of these traditional foods and an increase in obesity.

The aim of this work was to study the effect of the consumption of a traditional Mexican diet (TMD) in a model of diet‐induced obesity (DIO). To induce obesity, Sprague Dawley rats were fed a high‐fat diet+5 % of sucrose (HFD+S) in the water during 6 months. After that, rats were divided in 3 groups (n=6 per group): 1) obese rats fed HFD+S, 2) obese rats fed control diet, and 3) obese rats fed TMD during 3 months. Animals fed TMD decreased body weight, improved glucose tolerance and decrease serum and liver cholesterol triglycerides. These results could be explained by an increase in liver antioxidant enzymes, decrease in liver and brain MDA, increase in liver P‐AKT associated with insulin signaling, increase in energy expenditure and UCP‐1 protein in white adipose tissue, decrease in the expression of proteins involved in lipogenesis such as SREBP1c and FAS in liver, and by modifying the microbiota. These results indicate that the consumption of vegetable protein, functional foods, omega 3 fatty acids and bioactive compounds present in the TMD decreased significantly obesity induced by a high fat diet.
Ginsenoside Rg1 Inhibits Lipid Accumulation and ROS Production in 3T3‐L1 cells during Early Adipogenesis.
Authors: Koh Eun Jeoung , Jeon Hui Jeon , Seo Min Jung , Lee Yeon Joo , Ryu Su Jung , Hwang Ji Hyun , Choi Ji A , Lee Boo Yong
Published: June 2021

Adipogenesis is the process of adipocyte differentiation which increases lipid accumulation and ROS production. Many studies revealed that obesity is caused by excessive lipid accumulation and it leads metabolic diseases. Also, ROS production by oxidative stress induced insulin resistance which accompanies obesity and is a risk factor of type 2 diabetes. Ginsenoside Rg1 is one of the major components in panaxa ginseng, as a class of steroid glycosides, and triterpene saponins. Our result showed that Rg1 decreased lipid accumulation, C/EBPα, PPARγ, aP2 mRNA expression, C/EBPα, PPARγ protein expression and ROS production in a dose‐dependent manner. In addition, protein expression of early aipogenic factor C/EBPβ which induces C/EBPα, PPARγ was reduced during early adipogenesis. Our data suggests that Ginsenoside Rg1 inhibits lipid accumulation and ROS generation during early adipocyte differentiation. Depend on these data, we are planning to investigate anti‐oxidative factors.
Effect of Fresh Preparation and Industrial Processing on Stability of Potato Phytochemicals
Authors: Furrer A , Cladis D , Kurilich A , Hawkins G , Manoharan R , Ferruzzi M
Published: June 2021

Potatoes are a significant source of health promoting phytochemicals. However, reported content and process stability of phytochemicals in potato products is inconsistent. The objectives of this study were to determine changes in phytochemical content of select potato varieties through industrial processing and assess differences in phytochemical content between freshly “home” prepared (FP) and reconstituted, industrially processed (IP) white and sweet potato products. Commercial products included raw flesh w/o skin (FNS), blanched+frozen (BF), +microwaved (BFM), and blanched+par fried+frozen (BFF), +baked (BFFO) or +fried (BFFF). Anthocyanins and chlorogenic acids (CGA) were the main phenolic compounds in potatoes with 450, 600, and 63 mg CGA /100 g dw in flesh of purple, red, and white potatoes, respectively. For purple and red, CGA levels were significantly lower (P<0.05) in BFF, BFFO, and BFFF compared to FNS, BF, or BFM products. For white, CGA levels were lower (P<0.05) in all processed products compared to FNS, but no differences were observed between processing levels. Comparing FP to IP products, levels of CGA did not differ for classic fries or sweet potato fries, but were lower (P<0.05) in industrial baked and fried sweet potato wedges. Sweet potato β‐carotene was higher in industrial baked wedges (30 mg/100g dw) compared to fresh prepared (26 mg/100g) (P<0.05), but no other differences between FP and IP were found. These results suggest that industrial blanching, freezing, and microwaving have mild impact on phytochemical levels in raw potatoes compared to par frying and subsequent baking or frying. Additionally, industrial products compare favorably to freshly prepared products in recovery of phytochemicals.
Antioxidant and Neuroprotective Effects of Korean Fermented Food Doenjang
Authors: Kang Su Jin , Seo Ji Yeon , Kim Min Sun , Jo Gwang Rae , Ji Hyeon‐Kyeong , Erdenebileg Zolzaya , Jang Jin Ho , Shin Jung‐Hye , Jo Kye Man , Kim Jeong Hwan , Kim Jong‐Sang
Published: June 2021

Doenjang (fermented soybean paste) is one of the traditional fermented foods in Korea which has various physiological effects. In this study, we investigated the antioxidant and neuroprotective activities of doenjang prepared fresh and aged for 6 weeks. Freeze‐dried doenjang was extracted with 80 % (v/v) ethanol. The 80% ethanol extract of doenjang was subjected to determination of antioxidant activity by 2,2‐diphenyl‐1‐pycrylhydrazyl (DPPH) radical scavenging capacity, ferric ion reducing antioxidant power, 2,2'‐azinobis (3‐ehylbenzthiazoline‐6‐sulphonic acid) cation radical scavenging activity, and oxygen radical absorbance capacity assays. In addition, the neuroprotective effect of doenjang extract was investigated using MTT cytotoxicity and 2´,7´‐dichlorofluorescein diacetate (DCF‐DA) assays. It was found that the 80% ethanol extract of doenjang protected from neuronal cell death induced by ROS or glutamate. The underlying mechanism for neuroprotective effect of doenjang will be discussed. (supported by a grant from IPET (High Value‐added Food Technology Development Program, 2012, 112066‐3), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea)
Comparison of Different Tea Types on Osteoblast Activity
Authors: Nash Leslie , Ward Wendy
Published: June 2021

Many studies report that tea consumption is associated with higher bone mineral density in women. Polyphenols in tea may mediate such benefits. We examined if English Breakfast (EB) and Golden Monkey (GM) black tea, green tea (GT) or rooibos tea (RT) increased osteoblast activity in human osteoblast‐like cells. Preclinical studies typically use alcohol‐derived polyphenol extracts although tea for human consumption is prepared with water. We determined that polyphenol levels are higher with methanol versus water extraction (161 mg/g vs. 86 mg/g gallic acid equivalents, p < 0.001). Water extraction reduced polyphenol levels of GT, EB, and GM by more than 50%, whereas RT maintained 65% of its polyphenols. GT (117‐235 mg/g) and EB (121‐202 mg/g) had the highest polyphenol content, regardless of solvent (p < 0.001). For mineralization, the effect of water‐extracted polyphenols (1 or 10 µg polyphenols/mL media) were measured using the Alizarin Red assay. Cellular activity was examined by reduction of thiazolyl blue tetrazolium bromide (MTT). Two‐way ANOVA indicated a favorable effect of dose (1 or 10 µg/mL, p<0.001) and treatment (p < 0.001) on mineralization. 10 µg/mL tea extract produced less mineral than 1 µg/mL (132% vs. 157% of control, p < 0.001). Both doses of GT (p < 0.001, p < 0.001) and RT (p < 0.05, p < 0.01), but only 1 µg/mL of EB (p < 0.001) and GM (p < 0.001) increased mineralization. Both doses of GT (p < 0.05, p < 0.05) and RT (p < 0.001, p < 0.01) and 1 µg/mL of EB (p < 0.05) promoted greater cellular activity at 24 hrs. Neither dose of GM, nor 10 µg/mL of EB resulted in significant differences of MTT. Differing effects among tea types may be due to diverse polyphenol profiles. (Funded by NSERC Discovery Grant)
Effects of Flaxseed‐supplementation on DNA Methyltransferase and Obesity‐associated Gene Expression in Adipose Tissue from Obese Mice
Authors: Woodard Alycia , Rhee Yeong , Reindl Katie , Hall Clifford
Published: June 2021

Diet‐induced obesity (DIO) has been shown to increase DNA methyltransferase (DNMT) expression and DNMT binding at obesity‐associated genes. Natural compounds, such as flaxseed, have the potential to reverse obesity‐associated gene expression via the regulation of DNA methylation. The objective of this study was to determine the effect of the bioactive compounds of flaxseed on DNMT and obesity‐associated gene expression. Sixty C57BL/6J male mice were randomly assigned into one of the following diet groups and fed for eight weeks: 1) 45% Kcal fat; 2) 45% Kcal fat, 10% whole flaxseed; 3) 45% Kcal fat, 6% defatted flaxseed; 4) 45% Kcal fat, 4% flaxseed oil; and 5) 16% Kcal fat. DNMT1, DNMT3a, DNMT3b, leptin, and peroxisome proliferator‐activated receptor (PPAR)‐α expression levels in mouse adipose tissues were determined by real‐time PCR. DNMT1, DNMT3a, or leptin expression was significantly decreased in whole flaxseed, defatted flaxseed, flaxseed oil, or low fat diet groups compared to the high fat diet group. DNMT1 or DNMT3a expression was positively correlated with PPAR‐α and leptin expression in the defatted flaxseed, flaxseed oil, or low fat diet groups. The results indicate that bioactive compounds of flaxseed affect selected gene expression; however, further research is needed to identify the specific mechanisms regulating leptin or PPAR‐α expression during DIO development.
Reliability of Yeast Unclumping Assay, a Model for Testing Potentially Clinically Useful Reagents
Authors: Zem G , Cooperman B , Bahri F , Mahjoubi A , Warner N , Malekian L , Mirebrahimian R , Pistalu M , Khrayan L , Patel M , Pastrano K , Choi E , Baronian T , Gilani O , Cardenas A , Hambarsoomian A , Gomez D , Gallgos F , Holmes J , Vahdati V , Jorshari L , Grigorian P , Ohanessian K , Baum E , Majarian G , Aldzhyan K , Manasyan H , Allatabakhsh N , Oppenheimer Steven
Published: June 2021

In the project titled Hierarchy of Anti‐Clumping Salts in these proceedings, in 2525 separate trials by 45 independent investigators, a hierarchy of best to worst yeast unclumping agents was developed using 7 salts related to magnesium sulfate a known anti‐clumping agent. Here we test the long term reliability of this assay in 1404 trials by 29 mostly new investigators done 6 months after the original study. We focused on sodium sulfate, the best unclumper in the previous study. Using the same procedures described in the Hierarchy study, we found that after 60 min, when comparing the mean of percentage of single cells in experimentals vs controls, +20.0% single cells were found in the experimental samples. This is identical to the percent found a half year earlier, confirming the reliability of this assay. Here 18 of 20 groups of data with sodium sulfate, in 415 trials had a p value of less than 0.05 when comparing experimentals and controls. Other reagents were also examined here (magnesium carbonate and sodium bicarbonate) that were not effective unclumping agents. Identification of unclumping agents can be useful in anti‐thrombocytic, anti‐cancer, anti‐infection, anti‐biofilm as well as in industrial applications (support: Sidney Stern Memorial Trust, NIH RISE, NSF Presidential Award 0731633 and university funding sources).
ADIPOSE TISSUE EXTRACELLULAR MATRIX REMODELLING IN CANCER CACHEXIA
Authors: Alves Michele , Matos Neto Emidio , Maximiliano Linda , Alcantara Paulo , Otoch Jose , Batista Miguel , Seelaender Marilia
Published: June 2021

Cachexia is a paraneoplastic syndrome in which white adipose tissue (WAT) suffers profound wasting. WAT alterations that may lead to extracellular matrix (ECM) rearrangement. AIM: to assess the effects of cachexia upon ECM of subcutaneous adipose tissue (SAT) by examining the expression of collagens IIIα1 (COL3A1), VIα1 (COL6A1) and fibronectin (FN1).

Methods

Patients of the University Hospital were enrolled in 2 groups: Cachectic cancer (CACS,n=14)(gastrointestinal tumours) and Control (C, n=8) (hernia). One gram of SAT was collected in surgery and fixed in 4% formaldehyde and paraffin embedded. The sections (5µm) were stained by Picro Sirius red (PS) for polarized light microscopy, or employed for immunostaining, when slides were incubated with the antibodies overnight (4ºC). Gene expression analysis was performed by RTqPCR and normalised to reference gene 18s.

Results

PS stain demonstrated a strong presence of collagen in the SAT of CACS, and preserved ECM architecture was observed in C. Increased amounts of COL3A1 were found in CACS, despite no changes in gene expression. Immunostaining for COL6A1 was modified in CACS, with collagen aggregates in fibrotic areas and increased mRNA levels of COL6A1 (p<0.05). FN1 labelling was most evident in concomitance to augmented mRNA levels in CACS, comparing with C.

Conclusions

Cancer cachexia deeply affects SAT organization, causing accumulation of ECM components in the adipose pad leading to remodelling during cachexia. The increase of COL3A1, COL6A1 and FN1 may be directly related with the emergence of tissue fibrosis.
Ethics: ICB/USP(788);HU/USP(752/07). Funding: FAPESP (012/50079‐0).
Drug Dose Responses of 3D Cancer Spheroids in Microfluidics
Authors: Christ Theresa , Flint David , Boyd Marie , Zagnoni Michele
Published: June 2021

A major drawback in the development of new cancer therapeutics is the use of two‐dimensional (2D) monolayer cultures as models for solid tumours. A solution lies in the use of three‐dimensional (3D) multicellular aggregate models, known as spheroids or organoids. These mimic the interactions between cells within a tumour, the morphology and the microenvironment of tumours, representing a simplified but more physiologically relevant in vitro system for drug screening than that provided by 2D bioassays. In this study, we developed a novel microfluidic platform that allows the formation and long‐term culture of hundreds of multicellular cancer spheroids, retaining complete control over the number and the size of spheroids grown, the ability to deliver chemical compounds in a customisable concentration gradient format and the ability to recover the spheroids for off‐chip post‐processing after treatment. UVW cells, a human brain high‐grade glioma cell line, were used to create spheroids in an array format and were exposed to a microfluidic concentration gradient of cisplatin. Results showed that the growth of 3D glioma spheroids after treatment is reduced in a concentration‐dependent manner and that a dose response curve can be obtained from a single device. These findings demonstrate the suitability of the system for high‐throughput drug screening, combination therapeutics and the potential for use with biopsy samples.
Recurrence‐free Survival Among Patients With ER+/PR+/HER2‐ Breast Cancers is Predicted by Expression of the Estrogen Response Signature
Authors: Sandhu Rupninder , Parker Joel , Anders Carey , Coleman William
Published: June 2021

ER+/PR+/HER2‐ breast cancers are frequently associated with good outcomes, but some patients experience disease progression/recurrence. We examined gene expression patterns among ER+/PR+/HER2‐ breast cancers (with known molecular subtypes) to identify those that express a 583‐gene estrogen response signature (ERS), indicating the presence of an intact ER/PR pathway. Unsupervised cluster analysis of 205 ER+/PR+/HER2‐ breast cancers identified two clusters (85 and 120 breast cancers). Kaplan‐Meier analysis of recurrence‐free survival (RFS) did not identify significant differences between these clusters. Correlation analysis against an ideal ERS identified two subsets among ER+/PR+/HER2‐ breast cancers: 44 breast cancers were negatively correlated and 161 breast cancers were positively correlated with this signature. Kaplan‐Meier analysis of RFS revealed a statistically significant difference between these patient subsets (P=0.0048), suggesting that ER+/PR+/HER2‐ breast cancers that express the ERS are associated with better RFS than those lacking this signature. Molecular subtypes of ER+/PR+/HER2‐ breast cancers that do not express the ERS include 5 basal‐like, 5 claudin‐low, 7 HER2‐enriched, 23 Luminal B, but only 2 Luminal A breast cancers. Among ER+/PR+/HER2‐ breast cancers lacking the ERS 28/44 (64%) relapsed, compared to 44% relapse among all other ER+/PR+/HER2‐ breast cancers. Lack of expression of the estrogen response signature predicts more aggressive disease among ER+/PR+/HER2‐ breast cancers.
CTBP‐BRCA1 modulates Epithelial Cell Fate in Breast Cancer
Authors: byun Jung , Gardner Kevin
Published: June 2021

Epithelial cells retain incredible plasticity. They can differentiate and self‐renew during embryonic development. The epithelial cells can lose their typical epithelial features and change to a mesenchymal state by activating epithelial mesenchymal transition (EMT), whose process is critical for breast cancer metastasis. BRCA1 is tumor suppressor gene that represses malignant transformation by safeguarding the fidelity of DNA replication chromosomal segregation and has been demonstrated to play a possible role in mammary differentiation. C‐terminal binding protein (CTBP) is a dimeric nuclear protein and an epigenetic regulator that is activated in the presence of NADH to recruit and target a variety of histone modifying complexes and transcriptional regulators to chromatin, thus providing a defined mechanism through which carbohydrate metabolism can drive epigenetic regulatory events. We have recently shown that CTBP drives epithelial mesenchymal transition (EMT), genome instability, and the acquisition of stem cell tumor‐initiating cell‐like pathways in breast cancer.

Here we demonstrate that CTBP and BRCA1 competitively regulate epithelial features including epithelial polarity and lumen development in MCF10A 3D gland formation assays. Loss of BRCA1 appears to perturb mammary epithelial differentiation whereas CTBP depletion enhances epithelial differentiation. These findings define dynamic yet antagonist roles for CtBP and BRCA1 in mammary gland growth and development.
Therapeutic Benefit of Chemotherapy and Hormone Therapy Among Estrogen Response Signature‐Positive ER+/PR+/HER2‐ Breast Cancers
Authors: Sandhu Rupninder , Parker Joel , Anders Carey , Coleman William
Published: June 2021

ER+/PR+/HER2‐ breast cancers are frequently associated with good outcomes, but some patients experience disease progression/recurrence. We examined disease relapse and relapse‐free survival (RFS) as measures of response to chemotherapy (Chemo), hormone therapy (HT), and Chemo+HT among 182 ER+/PR+/HER2‐ breast cancers (with known molecular subtype) that differentially express a 583‐gene estrogen response signature (ERS). 25 patients received Chemo, 13 received HT, 20 received Chemo+HT, and 124 patients received no adjuvant therapy (surgery only). Overall, ERS+ ER+/PR+/HER2‐ breast cancers have better RFS (62/142, 43% relapse irrespective of therapy) versus ERS‐ ER+/PR+/HER2‐ breast cancers (26/39, 67% relapse irrespective of therapy). Regardless of ERS status, 29/58 (50%) treated patients relapse and 60/124 (48%) untreated patients relapse. However, response to therapy varies according to ERS status among treated patients. ERS+ ER+/PR+/HER2‐ breast cancers show benefit of HT or Chemo+HT – 45/96 (47%) untreated relapse and 10/19 (52%) Chemo relapse versus 3/9 (33%) HT relapse and 4/18 (22%) Chemo+HT relapse. ERS‐ ER+/PR+/HER2‐ breast cancers do not show benefit of Chemo, HT, or Chemo+HT – 15/28 (54%) untreated relapse and 11/11 (100%) treated relapse (irrespective of adjuvant modality). These results suggest that there is value in knowing ERS when treating ER+/PR+/HER2‐ breast cancers.
Cells and Molecules of Cancerous Neurogenesis
Authors: Steindler Dennis
Published: June 2021

Cancer stem‐like cells can be isolated and expanded using clonogenic sphere cultures from a variety of tumor types, including gliomas, osteosarcoma, colon and prostate cancer (Ignatova et al., 2002; Gibbs et al., 2005; Levings et al., 2009; Carpentino et al., 2009; Bae et al., 2009). In primary brain gliomas, astrocytes, microglia and stem/progenitor cell interactions, mediated by unique expressions of developmentally regulated molecules including extracellular matrix (“ECM”) that help build functional neural circuitry during development and neuropoiesis, together may also contribute to aberrant cell genesis during brain tumorigenesis (Silver and Steindler, 2009). There are distinctive roles for these elements as well during invasive versus non‐invasive glioma using both in vitro bioassays and patient‐derived xenotransplanted human glioblastoma (“GBM”) stem‐like cells (Silver et al., 2013; Ignatova et al., 2002; Siebzehnrubl et al., 2013). Microglia and their interactions with astrocytes and astrocytic cancer stem cells together regulate brain tumor organization and invasion. Gliotypic cells that transiently adopt tumorigenic behaviors during normal neurogenesis (Walton et al., 2009) may underlie gliomagenesis via their exposure to aberrant microglial signals, including inflammatory molecules that may be transmitted cell to cell via exosomes and that are targets of new combination therapies. Supported by NIH Grant NS055165, and the McKinney, Maren and Thompson Regenerative Medicine Fund.
Diabetic Ketoacidosis Alters Plasma Levels of Matrix Metalloproteinases and PMN‐Specific Elastase in Children
Authors: Woo Martin , Cepinskas Gediminas , Clarson Cheril , Fraser Douglas
Published: June 2021

Introduction

Diabetic ketoacidosis (DKA) is associated with cerebrovascular complications in children such as vasogenic edema. Circulating matrix metalloproteinases (MMPs), their endogenous inhibitors (TIMPs), and other proteases may contribute to loss of cerebrovascular endothelial cell junctional integrity. Our aim was to determine the effects of acute DKA on systemic MMP/TIMP levels, as well as polymorphonuclear neutrophil(PMN)‐specific elastase.

Methods

Plasma was obtained from children ages 4‐17 years old, with type‐1 diabetes, either in an acute DKA state or insulin‐controlled (CON). DKA and CON samples were age‐ and sex‐matched for all experiments. Gelatin zymography was used to assess gelatinolytic activity. ELISA‐based methods were used to assess plasma concentration of proteins.

Results

DKA plasma showed significantly increased MMP‐9 activity(P<0.01) and decreased MMP‐2 activity(P<0.001), compared to CON plasma. MMP‐9 activity in DKA plasma was inversely correlated with pH(P<0.05). DKA plasma had increased circulating levels of MMP‐3(P<0.05), MMP‐8(P<0.001) and TIMP‐4(P<0.01). In addition, a significant increase in circulating levels of PMN elastase(P<0.001) was detected in DKA plasma.

DKA is associated with dynamic changes in plasma levels of MMP‐2, MMP‐3, MMP‐8, MMP‐9 and TIMP‐4, as well as PMN elastase, which may contribute to loss of blood‐brain barrier integrity.