bluuebunny/crossref_metadata_2025 huggingface dataset:
Structural Modeling of the C‐terminal Domain of Ptf1a Bound to Mammalian Rbpj and Rbpjl
Authors: Osborn Nicholas , Mayberry Brett , Fein Zachary , MacDonald Raymond , Coats Ward
Published: June 2021

As a means to introduce biomedical research to high school students, the MacDonald laboratory at UT Southwestern Medical Center at Dallas has engaged students from Hillcrest High School in an ongoing research program to study the structure and function of a mammalian transcription factor complex. Ptf1a is a transcription factor that is crucial to the development of the embryonic pancreas. Ptf1a acts within the context of a trimeric complex composed of three DNA‐binding subunits: a common E‐box binding protein (E12/47, HEB, or TCF12), Ptf1a, and either Rbpj or Rbpjl. The Rbpj form of the complex (PTF1‐J) is required for the early stage of pancreatic development, whereas the Rbpjl form (PTF1‐L) is necessary for acinar development. Tryptophan 298 located in a conserved sequence (C2) near the C terminus of Ptf1a is required for the recruitment of Rbpj into the PTF1‐J complex. The binding of the RAM domain of Notch‐IC to a hydrophobic pocket in Rbpj involves residues in a conserved hydrophobic, tryptophan, hydrophobic, proline motif that is also present in the C2 sequence of Ptf1a suggesting a similar hydrophobic pocket binding interaction for tryptophan 298. We have modeled the C2 sequence of Ptf1a onto the crystal structure of the RAM domain of Notch‐IC bound to Rbpj to characterize this binding interaction and to generate structure models for the complex of C2 with mammalian Rbpj and Rbpjl.
Requirements of non‐heme iron and heme iron in the nematode, Caenorhabditis elegans
Authors: Zhao Yang , Lu Nancy , Belo Panfilo , McProud Lucy
Published: June 2021

A basal medium was prepared with both non‐heme iron and heme iron eliminated from C. elegans Maintaince Medium. After being pre‐cultured in this medium, C. elegans were inoculated into a 3×5 factorial design of experimental media, consisting of 5 levels of non‐heme iron (0, 0.0672, 0.336, 8.40, 210 μg/ml) and 3 levels of cyt c (0, 50.0, 500 μg/ml). The population growth was counted at the 28th Day, and iron uptake and % iron absorption of nematodes were analyzed by Atomic Absorption Spectrophotometry. At 0 μg/ml iron and 0 μg/ml cyt c, the population growth was very poor. At 0 μg/ml cyt c, as iron level increased to 8.40 μg/ml, the nematode population increased to 5.98±0.53×103 Nema/ml. As iron concentration further increased, significant declines in nematode population were observed, indicating iron toxicity. At 0 μg/ml iron, as cyt c level increased, the population growth was greatly increased. The highest population growth was achieved at 8.40 μg/ml iron and 500 μg/ml cyt c, suggesting an additive requirement of non‐heme and heme iron. The iron uptake increased when either non‐heme or heme iron levels increased in the media. At 0 μg/ml cyt c, the % iron absorption decreased with the increase of iron level in the media. This findings was in accordance with the “mucosal block” theory. At 50.0 and 500 μg/ml cyt c, the % iron absorption greatly increased, suggesting heme iron greatly enhanced non‐heme iron absorption in C. elegans.
γ‐Tocotrienol inhibits IL‐6 by suppression of C/EBPβ expression and NF‐κB signaling in lipopolysaccharide‐stimulated macrophages
Authors: Wang Yun , Jiang Qing
Published: June 2021

Interleukin‐6 (IL‐6) is a proinflammatory cytokine that contributes to the pathogenesis of inflammatory diseases such as arthritis. Here we showed that gamma‐tocotrienol (γ‐TE), a natural form of vitamin E, inhibited LPS‐stimulated IL‐6 production in RAW264.7 macrophages. Mechanistic investigation suggested that NF‐κB, but not JNKs, p38 or ERK was required for IL‐6 production in RAW264.7 cells. LPS caused transient activation of NFκB at 10 min presumably via Toll‐like receptor (TLR), while led to stronger activation at 60 min, as indicated by phosphorylation of IκBα, possibly by LPS‐induced cytokines. Interestingly, γ‐TE inhibited p‐IκBα at 60 min but not 10 min, suggesting that γ‐TE may inhibit cytokine‐ but not TLR‐mediated NF‐κB activation. Cytokine array probing a broad range of cytokine formation in response to LPS revealed that TNFα was likely the one that caused the delayed NF‐κB activation, which was confirmed by western blot. γTE treatment inhibited TNFα‐triggered p‐IκBa. However, TNFα alone or its combination with IFN‐γ failed to stimulate IL‐6, indicating that NF‐κB is required but not sufficient to induce IL‐6. We then examined potential effects of LPS and TNFα on C/EBPβ, a transcription factor known to promote NF‐κB‐mediated IL‐6. Results showed that LPS but not TNFα induced C/EBPβ expression. Consistently, C/EBPβ expression was positively associated with IL‐6 formation and knockdown of C/EBPβ by siRNA led to decrease of IL‐6. Importantly, γTE inhibited LPS‐stimulated C/EBPβ expression and led to increased expression of GADD153/CHOP10, a negative regulator of C/EBPβ. Our studies demonstrate that γ‐TE inhibited C/EBPβ and NF‐κB pathways, both of which are important regulators of inflammation, and therefore provide mechanisms of the antiinflammatory actions of this vitamin E form.
Effect Of A Fruit And Vegetable Juice Concentrate (FVJC) Vs. Medical Nutrition Therapy Alone On Metabolic Syndrome Components In Obese Children: A 6 Month Pilot Double Blind Placebo‐Controlled Study
Authors: Canas Jose Atilio , Damaso Ligeia , Altomare Astrid , Hossain Jobayer , Balagopal Babu
Published: June 2021

Epidemiologic studies report lower β‐carotene (BC) and retinol (ROL) in obese vs. non obese children, and suggest this likely predisposes to the metabolic syndrome (MS). We determined the effect of a 6‐month FVJC vs. Medical Nutrition Therapy (MNT) on the prevalence of MS based on 4 different criteria (MS1–Cook; MS2 and MS3‐Lambert; MS4‐de Ferranti) in 11 normal weight, 6 overweight and 22 obese pre pubertal children in a prospective randomized placebo‐controlled trial. Waist circumference, fasting glucose, insulin, triglyceride, HDL and systolic blood pressure (SBP) for age, sex and height (NHPBEP) were determined. Daily total kilocalories and average intake of vitamin A in retinol equivalents (VARE) or BC were derived from FFQ. ROL, BC, HOMA‐IR and QUICKI were determined at 0, 3 and 6 months. The prevalence of MS (MS4 criteria) decreased by 16 % at 3‐months (χ2 =6.004 p=0.014) in the FVJC vs. MNT but this effects was lost at 6 months; no significant change in HOMA‐IR or QUICKI; BC but not ROL correlated with a decrease in HOMA‐IR (χ2 = −0.393 p= 0.03) and an increase in QUICKI (χ2 =0.550 p=0.002), no correlation between intakes of VARE or BC and BC. Taken together, the increase in BC and reduction in MS (MS4 criteria) in FVJC vs. MNT suggest a potential role for BC in the regulation of insulin sensitivity in obese children. Population‐wide studies are warranted to determine the significance of this association.
Maternal dietary choline deficiency alters angiogenesis in fetal mouse hippocampus
Authors: Mehedint Mihai George , Craciunescu Corneliu N , Zeisel Steven H
Published: June 2021

We examined whether maternal dietary choline modulates angiogenesis in fetal brain. Pregnant C57BL/6 mice were fed either a choline‐deficient (CD), control (CT), or choline‐supplemented diet (CS) from days 12 to 17 (E12‐17) of pregnancy and then fetal brains were studied. In CD fetal hippocampus, proliferation of endothelial cells (EC) was decreased by 32% (p < 0.01 vs. CT or CS) while differentiated EC clusters (expressing factor VIII related antigen (RA)) increased by 25% (p < 0.01 vs. CT or CS). These changes were associated with > 25% decrease in the number of blood vessels in CD fetal hippocampus (p < 0.01 vs. CT and CS), with no change in total cross‐sectional area of these blood vessels. Expression of genes for the angiogenic signals derived from both endothelial and neuronal progenitor cells (NPC) was increased in CD fetal hippocampus VEGF C (Vegfc), 2.0‐fold, p < 0.01 vs. CT and angiopoietin 2 (Angpt2), 2.1‐fold, (p < 0.01 vs. CT)). Similar increased expression was observed in NPC isolated from E14 fetal mouse brains and exposed to low (5 μM), CT (70 μM), or high choline (280 μM) media for 72 h (low choline caused a 9.7‐fold increase in relative gene expression of Vegfc (p < 0.001 vs. CT and high) and a 3.4‐fold increase in expression of Angpt2, (p < 0.05 vs. CT and high). ANGPT2 protein was increased 42.2% (p < 0.01). Cytosine‐phosphate‐guanine dinucleotide islands in the proximity of the promoter areas of Vegfc and Angpt2 were hypomethylated in low choline NPC compared to CT NPC (p < 0.01). We conclude that maternal dietary choline intake alters angiogenesis in the developing fetal hippocampus. This work was supported by Grants from the National Institutes of Health AG09525, DK55865, and DK56350 (to S.H.Z.) and Pilot Grant ES010126‐08 (to M.G.M.)
Postprandial metabolism of EPA and DHA is affected by the normal menstrual cycle in young females
Authors: Silva Iris Lima , Torres Alexandre Guedes
Published: June 2021

Our aims were to investigate the postprandial metabolism of EPA and DHA during the follicular (FP) and luteal (LP) phases of the menstrual cycle of young females. Fourteen volunteers (21 to 36 years) were studied during the FP (3 to 9 d) and the LP (17 to 25 d) of their menstrual cycles. We collected blood samples after an overnight fast at baseline and at regular time intervals after the intake of a standardized meal: 870 kcal, 62% lipids, 29% carbohydrates, 2 g of fish oil, containing 360 mg EPA and 240 mg DHA. Blood samples were taken with 60 min intervals for up to three hours. The fatty acid composition of erythrocytes and total plasma were determined by gas‐chromatography. The apparent absorptions of EPA and DHA were significantly different (p< 0.05) between the follicular and luteal phases (average ± SD): 751,07 ± 681,19 and 110,30 ± 734,33, and 1938,94 ± 2831,34 and 1294,61 ± 2959,25, respectively. The nutritional status of DHA was positively associated with the apparent absorption of EPA, but not of DHA itself. DHA levels both in erythrocyte and in total plasma lipids presented significant (p< 0.05) positive correlations with the apparent absorption of EPA. Results indicate that the hormonal milieu, naturally occurring during the menstrual cycle, might affect the postprandial metabolism of DHA and EPA, and also that DHA status might affect n‐3 fatty acid absorption. Financial Support: FAPERJ, CNPq, CAPES (Brazil).
Conjugation of Rare Sugar with Amino acids by the Maillard Reaction
Authors: SHIRAI YOKO , SASAKI HITOMI , KAMIMURA WATARU
Published: June 2021

D‐psicose is thought to be an ideal substitute for sucrose for food products due to its sweetness, easy‐processing, and functional properties (non caloric and low glycemic response). In the present study, the effects of amino acid on the formation properties of D‐psicose were analyzed to find a better utilization of D‐psicose aerated food systems. The effects of reaction temperature, molecular ratio, and the number of amino group (lysine or glycine) on the conjugation of D‐psicose with amino acids were examined. It was found that under the condition of excess amino acids, rare sugar was produced more efficiently; the type of amino acid had no influence on the conjugation of rare sugar.
Effect of different meal timing on postprandial energy expenditure and sympathetic nervous system activity in adult Japanese men
Authors: Taira Takuya , Fujii Hisao
Published: June 2021

Objective

The objective of this study was to compare changes in EE (Energy Expenditure ) at breakfast, lunch, and supper using the indirect human calorimeter (IHC) for 24 hours in order to clarify the effect of difference in meal timing on postprandial EE.

Methods

We measured EE, LF/HF(Low Frequency/ High Frequency), HF(High Frequency) in adult Japanese men (n=13, 21.0 ± 0.3 years). EE was measured using the IHC for 24 hours. The activity of sympathetic nervous system was assessed by real‐time analysis system of heart rate fluctuation.

Results

Postprandial EE at supper time was significantly higher than that at breakfast time during the first 60 minutes after ingestion (p < 0.05) and that at lunch time during the first 50 minutes after ingestion (p < 0.05). However, preprandial EE was also significantly higher at supper time than those at breakfast time and lunch time (p < 0.05). Preprandial LF/HF was significantly higher at lunch time than at breakfast time (p < 0.05). The time course of LF/HF was significantly lower at supper time than at breakfast time at periods of 10, 30, and 80–90 minutes after ingestion (p < 0.05).

Conclusion

The increase in EE induced by intake of breakfast and lunch may affect preprandial EE at supper time. Food intake may be one of the factors in the diurnal rhythm of EE.
Comparison of in vitro fermentation characteristics of nondigestible oligosaccharides using fecal inocula from breast‐fed versus formula‐fed infants
Authors: Boler Brittany Vester , Rossoni Serao Mariana C. , Faber Trevor A. , Chow JoMay , Fahey George C.
Published: June 2021

Galactooligosaccharides, gum arabic, HP inulin, and selected human milk oligosaccharides were incubated (37°C) in anaerobic semi‐defined medium with fecal inocula obtained from healthy breast‐fed (BF)(n=4) and formula‐fed infants (FF)(n=4). Samples were collected over 12 h. pH declined (P=0.01) to a greater extent with FF versus BF, particularly at 6 h (P=0.03). Acetate production tended to be greater (P=0.10) with FF infants, with differences detected at 6 and 12 h (P<0.03). Propionate production within diet was affected by time (P<0.01), with greater production in FF compared to BF infants at 6 and 12 h. FF also resulted in greater (P=0.03) production of butyrate at 12 h. Both BF and FF accumulated significant lactate over time, but diet had not effect on lactate production. Total short‐chain fatty acid production was greater (P=0.04) with FF compared to BF, but this difference was only noted at 6 and 12 h. Quantitative PCR indicated lactobacilli populations in FF increased while BF infants decreased compared to baseline, with differences reached at 3 and 6 h (P<0.01). Bifidobacteria populations tended to be greater (P=0.09) in FF compared to BF by 12 h. In conclusion, FF fermented oligosaccharides more rapidly than those from BF, and the bifidogenic effect appeared to be more pronounced with the FF. This research was supported by Abbott Nutrition.
Ex vivo stimulation of neonatal porcine peripheral blood mononuclear cells with oligosaccharides found in human milk
Authors: Comstock Sarah S , Wang Mei , Hester Shelly N. , Li Min , Donovan Sharon M.
Published: June 2021

Infant formulas lack the complex mixture of oligosaccharides (HMO) found in human milk, which may be pivotal to the development of the neonatal microbiota and immune system. Few comprehensive analyses of the direct effects of HMO on immune cells from neonates have been undertaken. Herein, peripheral blood mononuclear cells (PBMC) isolated from 10 and 23 day old (do) sow‐reared pigs were stimulated with HMO (2FL, 3FL, 3SL, 6SL, SL, LNFPIII, LnNT, sLeX and HMO isolated from human milk [iHMO]) for 72h. T cell phenotype, cytokine production and proliferation were measured by flow cytometry, ELISA and 3‐H thymidine incorporation. Of the HMO tested, none had significant effects on any parameters examined. However, developmental differences were found. PBMC from 23do pigs produced more cytokines, had a higher proportion of CD8+CD3+ T cells and proliferated less than those from 10do pigs. PBMC from 23do pigs made more IL‐10 in response to iHMO, PHA or PHA+iHMO than PBMC from 10do pigs. Only PHA or PHA+iHMO stimulation induced IL‐4 secretion, and 10do pigs produced nearly twice as much IL‐4 as 23do pigs. Lastly, PBMC from 10do pigs proliferated 6–10 times more in response to PHA stimulation than did PBMC from 23do pigs. In summary, while appropriate developmental responses were observed, direct effects of HMO on porcine PBMC phenotype, cytokine production or proliferation ex vivo were not detected. (Supported by Abbott Nutrition)
Nutrition Status in Patients with Thalassemia Intermedia
Authors: Meksawan Kulwara , Thongkijpreecha Phurita , Kangsadalampai Oranong , Pongtanakul Bunchoo
Published: June 2021

Thalassemia patients, especially children are at risk of malnutrition. This study was conducted to assess nutritional status in children (5–15 years old) with thalassemia intermedia at Hematology Clinic, Siriraj Hospital. Twelve boys and 18 girls with thalassemia intermedia participated in this study. Nutritional status was evaluated by anthropometry, biochemical parameters and subjective global assessment (SGA). Dietary intake was also assessed by 24‐hour recall. All subjects appeared to have normal nutrition when assessed by SGA. However, according to % weight for age (W/A), 12 subjects (40.0%) were mildly malnourished, and 7 subjects (23.3%) were moderately malnourished. Based on % height for age (H/A), 26.7% and 10.0% of the subjects were mildly and moderately malnourished, respectively. The result of % weight for height (W/H) revealed that mild malnutrition was found in 46.7% of the subjects. Percentages of W/A, H/A, and W/H were correlated with tricep skin fold (TSF) (r = 0.70, 0.47, 0.50, respectively). Subjects had total energy intake lower than the recommendation. Serum total protein and albumin levels were in the normal range. This study demonstrated that thalassemia intermedia patients had various degrees of malnutrition. Combination of multiple methods in evaluating nutrition status may be necessary. This study was partly supported by Chulalongkorn University graduate school thesis grant.
Modeling the potential of lysine biofortification of maize to improve protein quality and reduce risk of protein inadequacy in Sub‐Saharan Africa
Authors: Suri Devika , Strutt Nicholas R. , Ghosh Shibani , Gunaratna Nilupa S.
Published: June 2021

Introduction

Populations dependent on starchy staple crops are at risk of protein inadequacy due to limited lysine content. Previous work found 31% risk of protein inadequacy in Sub‐Saharan Africa (SSA). We examined the potential of lysine biofortified maize to reduce protein inadequacy in countries with high maize dependence.

Methods

FAO food balance sheets and USDA nutrient data were used to determine mean nutrient supply and estimate the population risk of protein inadequacy (PRPI), adjusting for protein quality using the PDCAAS method. We modeled a 60% increase in lysine in maize to bring the amino acid score to 1 as per WHO/FAO 2008 adult protein requirements. The new PRPI was calculated based on 2005 food supply, assuming 100% substitution with biofortified maize.

Results

Lysine biofortified maize decreased PRPI by more than 10 percentage points in Mozambique (77% to 60%), Zimbabwe (43% to 28%), Togo (53% to 40%), Malawi (31% to 18%), Zambia (44% to 33%) and Lesotho (13% to 3%), equating to 10 million fewer people at risk for protein inadequacy. No other amino acid was limiting in the diet.

Conclusion

Lysine biofortification of maize has the potential to significantly reduce PRPI in countries with high dependence on maize. The model should be applied to individual‐level dietary data to assess PRPI in at‐risk populations.
Accuracy of stated energy contents of chain restaurant foods in a multi‐site study
Authors: Urban Lorien E , McCrory Megan A , Dallal Gerard E , Das Sai Krupa , Saltzman Edward , Weber Judith L , Roberts Susan B
Published: June 2021

New government legislation will mandate increased reporting of nutrition information by restaurants but the accuracy of such information is uncertain. We used bomb calorimetry to measure energy contents of 269 randomly selected foods from chain restaurants in three states and compared the measured values to restaurant stated values. Overall, measured energy was extremely variable but on average not different from stated values corrected for metabolizable energy factors (+7±43%, P=0.57). In foods obtained from sit‐down restaurants, items with low stated energy (<600 kcal) contained more energy than stated compared to those with high energy (≥600 kcal) (+5% vs. −8%, P=0.05). In addition, the 10% of foods that had the greatest discrepancy between measured vs. actual energy content were re‐sampled and again had a similar discrepancy (54±28% and 44±26%, respectively, both P <0.0001 compared to 0%). In summary, the stated energy values of restaurant foods are frequently inaccurate, and menu choices that are apparently appropriate for weight control tend to contain more energy than stated. These errors may hamper use of restaurant‐provided nutrition information for self‐monitoring of energy intake for control weight. Funded by Tufts University, Purdue University, Arkansas Children's Hospital Research Institute, Arkansas Biosciences Institute.
Food Consumption Patterns of Adults Following a Self‐selected Prescreening for Chronic Disease Risk
Authors: Haley‐Zitlin Vivian , Mukwevho Peter , Barron Felix , Bodine Ashby
Published: June 2021

This study evaluated food consumption choices of male and female individuals who had previously participated in a comprehensive screening for chronic disease risk. Participant results following the screening included a nutrition score, based upon the participant's eating habits (including consumption of low‐fat meals, high‐fiber foods, breakfast consumption, fast food/snacks and food guide recommendations). Many factors may influence dietary intake changes including knowledge of chronic disease risks. However, altered dietary intake behaviors have been variable. Dietary intake of participants for 12 months was assessed electronically by the Block Food Frequency Questionnaire, Nutriquest. Most subjects reported their ethnicity as Caucasian, 88%, followed by African America and Hispanic; average age, 53. All participants were employed. Reported health status was: excellent, 20%; very good, 47.5%; good, 30%; fair, 2.5%. Mean caloric consumption was 1780±512.4kcal. Macronutrient consumption was CHO (47.7%), protein (35.6%), fat (16.1%) with intake of MUFA>SFA>PUFA. Dietary cholesterol was 206 mg/day. Fiber intake of 20.6 mg/day was reported. Calculated BMI indicated most subjects were currently overweight/obese (55%). Results clearly indicate the importance of nutrition education and follow‐up of individuals who exhibit proactive health related behaviors.
Do breastfed and formula‐fed infants differ on hair manganese levels?
Authors: Rink Stephanie , Bartok Cynthia , Kordas Katarzyna
Published: June 2021

Manganese (Mn) toxicity is observed in children exposed to high Mn levels in water and in total parenteral nutrition. Infant formula contains 5–28 times as much Mn as breast milk, but is not widely studied as a source of Mn toxicity. The objective of this study is to determine the difference in hair Mn levels between infants who are predominantly formula‐fed (FF) or breast‐fed (BF), and explore other salient predictors of hair Mn levels. Infants 3–6 mo who were FF (n=5) or BF (n=30) were recruited. Water and hair Mn levels for mother and infant were determined by ICP‐MS analysis. Mean±SD (median) hair Mn for BF and FF infants was 2.21±2.77 (1.45) μg/mL and 2.76±1.80 (3.16) μg/mL, respectively. Group medians differed significantly (p=0.002). 50.0% of FF displayed elevated hair Mn levels (>3 μg/g), compared to 13.8% of BF infants. Mean water Mn concentration was 0.92±2.10 ng/mL for BF and 0.21±0.16 ng/mL for FF infants. Infant hair Mn was not correlated with water Mn concentration (p=0.32), sex (p=0.21) or age (p=0.25). Additional samples are being collected to provide more definitive results.
Inhibitory effect of t10 c12 conjugated linoleic acid (CLA) isomer on clozapine induced adipogenesis in 3T3‐L1 cells
Authors: Park Yooheon , Kim Younghwa , Lee Junsoo , Park Yeonhwa
Published: June 2021

Atypical (second generation) antipsychotics such as clozapine have become the primary choice of medication for treating psychosis. An unfortunate side‐effect of these medications is weight gain, a condition often associated with the development of metabolic disorders such as diabetes. Factors thought to facilitate this weight gain include increased food intake, reduced energy expenditure, and impaired pre‐adipocyte differentiation. CLA, specifically the t10 c12 isomer, has been known to suppress adipogenesis both in vitro and in vivo model. Thus we tested the effect of the t10 c12 CLA on clozapine induced adipogenesis in 3T3‐L1 cells. 3T3‐L1 pre‐adipocytes were differentiated by the addition of insulin (1μg/ml), dexamethasone (1μM), IBMX (10mM), and 10% FBS in DMEM medium. Cells were treated with or without clozapine (10 and 20μM) and/or t10 c12 CLA (50μM). Lipid accumulation was measured by Oil‐red O staining and intracellular triacylglyceride contents were analyzed. mRNA level of the lipid metabolism related transcription factors were measured by RT‐PCR. Cells treated with clozapine alone showed significant increase in lipid accumulation compared to control. However, increased lipid accumulation by clozapine was significantly prevented in cells treated with CLA. mRNA level of fatty acid synthase (FAS), acetyl‐CoA carboxylase (ACC), uncoupling protein‐2 (UCP‐2), and peroxisome proliferator‐activated receptor‐γ (PPAR‐γ) in the cells treated with CLA were significantly decreased by t10 c12 CLA with or without treatment of clozapine. These results suggest that CLA may be used to prevent fat accumulation associated with antipsychotic administration.
Anti‐hypertensive potential of ethylacetate extract from radish leaves in spontaneously hypertensive rats
Authors: Chang Moon Jeong , Jeong Da‐Hee , Park Jee‐Ho , Jun Young Hee , Cho Kangjin , Kim Sun‐Hee
Published: June 2021

The aim of this study was to investigate the effects of ethylacetate extract from Radish Leaves (EERL) on the blood pressure in spontaneously hypertensive rats. Rats were divided into four groups : 50% DMSO, two levels of EERL (30 mg/kg bw or 90 mg/kg bw) in SHRs, normotensive Wistar‐kyoto. Rats were orally administrated for 5 weeks. Systolic blood pressure, angiotensin‐converting enzyme (ACE) activities of serum, kidney and lung, tumor necrosis factor‐α (TNF‐α), nitric oxide (NO), anti oxidative enzyme activity and malonaldehyde level were measured. EERL tended to attenuate blood pressure by increasing serum NO, reducing TNF‐ α and pulmonary ACE activity in SHRs. EERL decreased lipid peroxidation and increased antioxidant effects in SHRs. EERL showed good antihypertensive potential and antioxidant effects at a dose of 30mg/kg bw, not at a dose of 90mg/kg bw. This is the first report on the in vivo anti‐hypertensive potential of ethyl acetate extracts from radish leaves for use as medicinal food for hypertension. The experimental design was approved by the KMUIACUC regarding the care and use of laboratory animal. (This research was supported by 15 agenda research project from KRDA)
The Effect of lecithin extracted from Japanese anchovy (Engraulis japonicas) on Blood Passage and Serum Lipid Profile in Rats
Authors: Back Hyang‐Im , Kim Sun‐Young , Park Soo‐Hyun , Park Eun‐Ock , Oh Mi‐Ra , Kim Min‐Gul , Jung Su‐Jin , Oh Hong‐Geun , Park Joeng‐Woo , Kim OkJin , Lee Seung‐Mi , Sim Jeong‐Eun , Chun Byung‐Soo , Chae Soo‐Wan
Published: June 2021

To investigate the effects of lecithin extracted from Japanese anchovy on blood passage and serum lipid profile were assessed in vivo. Sprague Dawley strain rats were divided into five groups of ten rats each; normal control, negative control and three dosages of lecithin treated groups (80, 200 and 500 mg/kg). Serum total cholesterol, triglyceride, LDL‐cholesterol, atherogenic index (AI) and cardiac risk factor (CRF) contents were significantly reduced in the lecithin treated groups compared to the negative control group (P<0.05). The level of serum serotonin in the lecithin treated groups was significantly reduced compared to the negative control group (P<0.05). The platelet aggregation was inhibited in lecithin treated groups. There was no significant difference compared to the negative control group. Blood passage time of the lecithin treated group showed a tendency to decrease compared to the negative control group, which was studied with micro channel array flow analyzer (MC‐FAN). Also, in the histopathological assay of aorta wall lumen, the cholesterol aggregation level in the lecithin treated groups was reduced compared to the negative control group. All these results suggest that Japanese anchovy‐extracted lecithin might have anti‐atherogenic effect via lipid profile improvement.
Effect of fruit and vegetable concentrates on endothelial function in persons with metabolic syndrome: A randomized, controlled, crossover trial
Authors: Njike Valentine , Ali Ather , Yazaki Yuka , Ma Yingying , Katz David
Published: June 2021

Dehydrated fruit and vegetable concentrates provide an accessible form of phytonutrient supplementation that may offer cardioprotective effects. This study assessed the effects of encapsulated fruit and vegetable juice powder concentrates on endothelial function (EF) and cardiac risk markers in subjects with metabolic syndrome (MS) in a randomized, double blind, placebo controlled crossover trial. 64 adults with MS were enrolled and received 8‐week sequences of two different blends of the concentrates and placebo. The primary outcome measure was change in EF 2hr after consuming a 75g glucose load, after 8‐weeks of daily consumption or 2hr after consumption of a single dose. Secondary outcome measures included plasma glucose, serum insulin, serum lipids, and weight. A single dose of fruit, vegetable, and berry mixtures appeared to preserve EF (p=0.0549). There were, however, no significant between group differences in EF with daily treatment for 8 weeks. No other significant treatment effects were discerned in glucose, insulin, lipids, and weight. Daily supplementation with fruit and vegetable concentrates did not improve EF or other cardiac risk measures in this population of adults with MS in whom baseline EF was fairly normal. A blend of fruit, vegetable, and berry mixtures appeared to mitigate adverse effects of an acute glucose load on EF, a finding which warrants further investigation.
Validation of Array‐based RNA Expression Profiles in Paraffin‐embedded Samples of Epstein‐Barr Virus‐related Malignancy
Authors: Tang Weihua , Banet Natalie , Muallem Hind , Sailey Charles J , Hu Zhiyuan , Ma Shidong , Kenney Shannon , Raab‐Traub Nancy , Gulley Margaret L.
Published: June 2021

Epstein‐Barr virus (EBV) can be found in malignant cells and its viral genes represent targets for therapy and markers of tumor burden. In this study, the Nanostring system was used to evaluate 20 viral and 73 human RNAs in 13 paraffin embedded cell lines and in 16 carcinomas with matched non‐malignant tissue. RNA was successfully extracted from all specimens, and 100ng of RNA was used for analysis. Three different RNA purification strategies were compared. Crude lysates of RNA produced similar expression profiles but at lower levels than purified total nucleic acid. In addition, 3 spiked synthetic ERCC RNAs were successfully measured. Human tissue samples lacking EBV infection by the EBER in situ hybridization assay were essentially negative for all of the EBV transcripts. Unsupervised clustering revealed distinct and credible patterns of expression in epithelial vs lymphoid cell lines, in infected vs uninfected tissues or cell lines, and in tightly latent vs permissive forms of viral infection. This validation data suggests that RNA expression profiling of paraffin embedded tissue is feasible, and that use of purified nucleic acid promotes quality assurance. The synthetic ERCC reagents, when added during specimen processing, provide a resource for assessing quality of stored RNA.
Expression of BCL‐6 and cyclin D in human colon carcinoma
Authors: Baidoo Cletus , Babbra Ramandeep , Pathan Muhammad , Quinn Tim , Herndon Betty , Molteni Agostino
Published: June 2021

Cyclin D, an upregulated gene of colorectal cancers is used as a diagnostic marker for colon carcinomas. BCL‐6 is a transcriptional regulator important for lymphocyte survival. Microarrays show BCL6 is seven‐fold upregulated in colorectal cancers compared to controls. We tested whether BCL‐6 could be used as a diagnostic marker in moderately differentiated colonic adenocarcinomas by comparing the proportion of BCL‐6 staining to Cyclin D control. With IRB approval, human neoplastic colorectal masses were obtained from surgical pathology specimens on 31 subjects. Antibodies for BCL‐6 and Cyclin D were tested in control tissues and on all sections with antigen baring, secondary and tertiary staining for both markers, n=62. Sections were graded by 2 pathologists unaware of the slides' identity. Data were also evaluated for inflammation, mucin, positive lymph nodes and polyp origin. Analysis by ANOVA included 4 groups, BCL‐6 positive, cyclin D positive, tumors positive for both or negative for both markers. Of the 100% surgically diagnosed cases, BCL‐6 was positive in 16% and cyclin D positive in 29%. BCL‐6 was significantly related to cases “mucinous” cases (p<0.001). Marked inflammation (p<0.01) was noted in tumors positive for both markers. This study attempted the validation of another biomarker for colon cancer. A need exists to predict treatment responsiveness. Sponsor: Sarah Morrison grant, UMKC
Rmi1 is essential for DNA damage response and embryonic development in mice
Authors: Guiraldelli Michel Farchi , Pezza Roberto
Published: June 2021

Suppression of Crossovers (CO) is critical for maintenance of genome integrity. CO may lead to loss of heterozygosity and CO between low‐copy repeats may account for many human disorders. Eukaryotes possess mechanisms that down regulate the number of CO. Indeed, BLM, the helicase mutated in Bloom syndrome, associates with topoisomerase IIIα and Rmi1 (RecQ‐mediated genome instability 1) to form a complex (the BLM dissolvasome) that efficiently control the number of CO by specifically resolving a key intermediate of recombination, the double Holliday junction (DHJ). To address the requirement of Rmi1 in mammals we generated knockout mice for this protein. Our results show that Rmi1 is required for normal embryonic development. Rmi1−/− knockout embryos die at 8.5 dpc, showing profound developmental defects reminiscent to those described for BLM−/− knockout mice. We obtained mouse embryonic fibroblasts from Rmi1−/− knockout and determined that these cells showed significantly reduced survival rates with respect to Rmi1+/− heterozygote and wild‐type controls. In this work, we will present a detailed characterization of developmental defects observed in Rmi1−/− knockout mice and study the DNA damage response of Rmi1 deficient cells. Our results will define how and when Rmi1 regulates the repair of DNA double‐stand breaks. This is important for understanding the mechanism by which somatic cells eliminate deleterious recombination products which are the root cause of genome instability and losses of heterozygosity.

Financial support: OCAST
Caloric Restriction Reduces Mammary Tumor Growth and Metastases
Authors: De Lorenzo Mariana S. , Baljinnyam Erdene , Vatner Dorothy E. , Abarzua Patricio , Vatner Stephen , Rabson Arnold B.
Published: June 2021

Caloric restriction (CR) is the most widely studied model of extending lifespan. Several studies have demonstrated that CR has beneficial effects on limiting tumor growth; however, less is known about CR effects on metastases and the extracellular matrix (ECM), which is important for tumor invasiveness. We investigated this in a mouse breast cancer model (4T1). Mice were fed with normal diet (ND) or 40% CR. After 5 weeks on diets, 4T1 cells were injected into BALB/c mice and diets continued for the remainder of the experiment. Mice under CR had a 23% reduction in body weight compared to ND mice (p<0.01). CR reduced tumor growth and weight (CR: 1.5±0.3 vs. ND: 3.5±0.4g; p<0.01). Tumors from CR mice demonstrated a 64% decreasein the cell proliferative index (p<0.01), lower microvessel density (p<0.05) and total vessel length (p<0.01) and increased apoptosis (p<0.05). CR reduced (p<0.05) the numbers and sizes of spontaneous and experimental (p<0.05) lung metastases. CR mice demonstrated lower MMP‐9 (matrix metalloproteinase‐9) activity (ND: 1.1±0.1 and CR: 0.7±0.1 ng/ug protein; p<0.05) and a 34% lower total collagen intra‐tumor volume fraction compared to ND mice, p<0.05. Our results suggest that CR reduces mammary tumor growth and its metastases, by increasing apoptosis, reducing proliferation and angiogenesis, while remodeling the ECM.
Granulovacuolar degeneration bodies of Alzheimer's disease resemble late‐stage autophagic organelles
Authors: Funk Kristen E , Mrak Robert E , Kuret Jeff
Published: June 2021

Granulovacuolar degeneration involves the accumulation of large, double membrane‐bound bodies within certain neurons during the course of Alzheimer's disease. Because of the two‐layer membrane morphology, it has been proposed that the bodies are related to autophagic organelles. To test this hypothesis, spatial colocalization of autophagic and endocytic markers with casein kinase 1 delta, a marker for granulovacuolar degeneration bodies (GVBs), was investigated in hippocampal sections prepared from post mortem Alzheimer's disease cases using double‐label confocal fluorescence microscopy. Results revealed that GVBs colocalized most strongly with LAMP1 (lysosome‐associated membrane protein 1) and CHMP2B (charged multivesicular body protein 2B), but weakly with lysosomal marker cathepsin D or early‐stage autophagy markers LC3 and p62. The resultant immunohistochemical signature suggests that GVBs do contain late‐stage autophagic markers, and accumulate at the nexus of autophagic and endocytic pathways. The data further suggest that failure to complete autolysosome formation may be an important correlate of GVB accumulation. This work was supported in part by the National Institute on Aging (AG14452, JK).
Chronic Caloric Restriction Improves Locomotor Activity in Aging Female Mice
Authors: Moore Marinda , DeLorenzo Mariana , Yan Lin , Chirieac Doru , Bravo Claudio A. , Vatner Dorothy E. , Vatner Stephen F. , Gelpi Ricardo
Published: June 2021

Caloric restriction (CR) is the most widely studied model of longevity. Aging induces alterations in behavior and reduced locomotion, i.e., bradykinesis, is a reliable biomarker of aging. CR, applied for a relatively short time (weeks to months) has been shown to improve bradykinesis, but it is not known what happens when the animals advance to old age. In order to explore the behavioral effects linked to long term CR and aging, we examined the behavior study of old (>24mo) female mice under CR since youth, with mice monitored for the same time on normal diets (ND). Average body weights fell over 2 years for CR mice (19 to 16g) and, as expected, increased for ND mice (18 to 25g). Beginning at 10 weeks of age, mice were fed either a normal diet (ND (equivalent to 10% CR)) or 40% CR and monitored up to 24 months of age. We used an open field cage system to store, replay, and analyze locomotor activity expressed as socially interactive or motile behavior. The mice under chronic CR displayed hyper‐activity (social and motile behaviors) as compared to ND mice. While comparing ND vs. 40% CR mice respectively, we detected an increase in stereotypy episodes (188 ± 13 to 458 ± 106 counts, P<0.05), movement time (658 ± 83 to 1559 ± 348 sec, P<0.05), and decrease in rest time (6540 ± 83 to 5637 ± 348 sec, P<0.05). These results suggest that increased locomotor activity is not only an early feature of CR, but persists with aging. This may not only be a biomarker for longevity, but also may be involved in the mechanism of longevity induced by CR.
Suboptimal release of CD34+/CD144+ cells in atherosclerotic patients in response to ischemia: role of plasmatic TGF‐B
Authors: Tejerina Teresa , Gonzalez‐Rocafort Alvaro , Navarro Jorge , Ramajo Marta , Hristov Mihail , Weber Christian , Martinez‐Gonzalez Jose , Gordillo‐Moscoso Antonio , Redondo Santiago
Published: June 2021

Objectives

To determine the number of CD34+/CD144+ cells in coronary artery bypass grafting (CABG) patients, compared to valvular patients and healthy controls, and the apoptotic effect of their plasma on peripheral blood‐derived angiogenic early outgrowth cells (EOCs) from healthy donors.

Methods

CD34+/CD144+ cell counts were assessed by cell cytometry from peripheral blood mononuclear cells. EOCs were separated by ficcoll gradient and cultured on fibronectin using microvascular medium. Protein expression in EOCs was determined by Western blot.

Results

We found a lower number of CD34+/CD144+ cells in CABG patients compared to valvular patients, but a higher number compared to controls. Being bypass or valvular had a strong and independent association with CD34+/CD144+ even after the P was adjusted by the number of comparisons. Plasma from CABG patients decreased apoptosis in cultured EOCs from healthy donors (day 7). This effect was abrogated by the transforming growth factor‐beta 1 (TGF‐b1) blockers SIS3 and SB‐431542. Of note, prior to apoptosis protection, plasma from bypass patients was able to upregulate the expression of both CD34 and CD144 at day 4 of isolation.

Conclusion

Bypass patients, which a high degree of ischemia and previous infarct rate, have a suboptimal release of CD34+/CD144+ despite a TGF‐beta‐mediated protective effect of their plasma on cultured EOCs.
Captorpil and losartan modulate vascular function differently in rats with type I diabetes mellitus
Authors: Sabra Ramzi , Fakhry Josiane
Published: June 2021

We investigated the influence of treatment with either captopril (50 mg/kg/day) or losartan (5 mg/kg/day) over two weeks in diabetic and control rats, on responses of the thoracic aorta to various vasoactive substances. Studies were conducted 4 weeks after induction of diabetes with streptozotocin (85 mg/kg i.v.) or its vehicle for control rats, and the drugs were infused over the last 2 weeks of that time period. Responses to acetylcholine (Ach), phenylephrine (PE), and SNAP (a NO donor) were obtained. Aortic rings from diabetic rats had decreased responses to Ach, enhanced responses to SNAP (which were corrected by DMPPO – a phosphodiesterase 5 inhibitor), and unchanged responses to PE. Captopril corrected the reduced response to Ach, did not affect the enhanced responses to SNAP and rendered the response to PE significantly lower in the rings from diabetic rats relative to controls. In contrast, treatment with losartan did not affect the responses to Ach, which remained depressed in diabetic rings, corrected the enhanced responses to SNAP in diabetic rings back to control levels, and did not influence the responses to PE. These results suggest that whereas captopril treatment appears to enhance vasodilator mechanisms in aortic rings from diabetic rats, losartan treatment does not have that effect and is not protective. However, losartan treatment appears to enhance responses to SNAP, possibly through decreasing phosphodiesterase 5 activity, while captopril does not have that effect. The mechanisms responsible for the differences between the two drugs need further investigation.
Tick‐borne pathogens in Winona County: Borrelia burgdorferi and Anaplasma phagocytophilum in Ixodes scapularis
Authors: Wallerich Lukas Roseman , Minnerath Jeanne
Published: June 2021

Tick‐borne pathogens can cause a number of harmful human diseases, such as Lyme disease from Borrelia burgdorferi and Anaplasmosis from Anaplasma phagocytophilum. Ixodes scapularis, commonly known as the deer tick, is a well‐established tick species within Winona County that is known to be a competent vector for these pathogens. The purpose of this study was to collect deer ticks in Winona County and assess the prevalence of B. burgdorferi and A. phagocytophilum. Deer ticks were collected from five sites (100 ticks per site), and DNA was extracted and analyzed for the presence of either pathogen using real‐time polymerase chain reaction (rtPCR). On average, 40% of the ticks were infected with B. burgdorferi, while 16% of the ticks were infected with A. phagocytophilum. These results are consistent with data collected from other regions within southeast Minnesota. Interestingly, 7% of the ticks were co‐infected with both pathogens. This value was higher than that observed in previous studies (2% co‐infection). Surveillance studies such as this are important to monitor tick populations and their pathogens and to insure that the public is aware of the prevalence of these tick‐borne pathogens. Funding for this research was provided by the National Biological Honors Society, Beta Beta Beta; Gundersen Lutheran Medical Center, La Crosse, WI; and Saint Mary's University of Minnesota, Winona, MN.
Ethanol extracts of Antrodia camphorata inhibit invasion and migration of human non‐small cell lung carcinoma cells
Authors: Chen Ying‐Yi
Published: June 2021

Antrodia camphorata (AC) is a fungus native to Taiwan which is believed to be effective in preventing diseases. Recent reports demonstrate that AC induces apoptosis of various tumor cells. In this study we investigated how ethanol extract of AC affect metastatic activity in CL1‐0 and CL1‐5 lung cancer cells. The underlying mechanism was also investigated via Western blotting. Our results demonstrated that ethanol extract of AC dose and time‐dependently inhibited the migration and invasion of CL1‐0, CL1‐5 cells under non‐cytotoxic concentrations. However, ethanol of AC demonstrated more inhibition on CL1‐5 than CL1‐0. When treating with AC, MMP‐9 and MMP‐2 activity were reduced in the gelatin zymography assay. By Western blot analysis, treatment with AC decreased phosphatidylinositol3‐kinase/AKT, MMP‐2 and MMP‐9 expression in CL1‐0 and CL1‐5 cells. AC also exerted an inhibitory effect on the phosphorylation of extracellular signal‐regulated kinases 1 and 2 and inhibition of activation of nuclear factor kappa B, c‐Fos, and c‐Jun. So, according to these data, the growth inhibition of lung cancer cells by AC was mediated by induction of metastasis and cell proliferation related proteins. As shown by the above results, Antrodia camphorata may be a powerful candidate in developing preventive agents for cancer metastasis.
Policyclic Pyridinones and Pyrimidinones and their Antimicrobial Activity
Authors: Damoni‐Rexha Dafina , Breznica‐Selmani Pranvera , Hoti Ramiz , Begolli Yllka , Troni Naser , Thaçi Veprim
Published: June 2021

Novel Pyridinones and pyrimidinones are synthesized by cyclic condensation of 2‐aminopyridines, respectively 2‐aminopyrimidines and corresponding acids. Condensation of 2‐amino‐4,6‐dimethylpyridine 1a and chloroacetic acid 2a gave corresponding imidazo‐[1,2,4]‐4,6‐dimethylpyridin‐3(2H)‐one 3a, whereas cyclocondensation of 1a and malonic acid 2b, gave 6,8‐dimethylpyrido‐[1,2‐a]‐pyrimidin‐2,4‐dione 3b. By condensation of 2‐amino‐4‐chloro‐6‐hydroxypyrimidine 1b and chloroacetic acid 2a, 7‐chloro‐5‐hydroxy‐2H‐imidazo‐[1,2‐a]‐pyrimidin‐3‐one 3c was synthesized, whereas cyclocondensatiuon of 1b and malonic acid 2b gave 8‐chloro‐6‐hydroxypyrimido‐[1,2‐a]‐pyrimidin‐2,4‐dione 3d. The antimicrobial activity of products 3(a‐d) were tested and results were submitted for their activities against E. coli, S. aureus and Candida albicans. Compounds 3c and 3a showed emphatic bactericide activity against E. coli. Considerable activity against S. aureus exhibited compounds 3b and 3c, whereas 3c and 3d exhibited the greatest activity against Candida albicans. In general there was observed that increasing of concentration causes greatest antimicrobial activity.