bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of brain nitric oxide in cardiovascular control of frogs
Authors: Zena Lucas A. , Gargaglioni Luciane H. , Bícego Kênia C.
Published: June 2021

The aim of the present study was to determine if nitric oxide (NO) acting on the brain of frogs presents an inhibitory effect on arterial blood pressure (ABP) and heart rate (HR) by reducing the sympathetic activity dependent on α and β adrenergic receptors. Thus, body temperature and ABP was measured by a telemetry device implanted into the abdominal cavity of american bullfrogs (250–350 g BW), with the catheter of the device inserted into the left aortic arch for ABP measurements. A guide cannula was implanted into brain lateral ventricle for injections of L‐NMMA (non selective NO synthase inhibitor) or mCSF (mock cerebrospinal fluid). A catheter was inserted into the femoral vein for injection of adrenergic antagonists (prazosin: α1; sotalol: β) and agonists (phenylephrine: α1; isoproterenol: β) or Ringer solution. Animals (n= 6) were maintained at 25oC during experiments. L‐NMMA increased (P< 0.05) ABP by 7.5±1.3 mmHg compared to mCSF (−0.5±0.7 mmHg) and did not change HR (delta HR: −1.37±1.8 bpm and −0.9±0.6 bpm, L‐NMMA and mCSF respectively). The pre‐treatment with prazosin, but not with sotalol, attenuated the hypertensive effect of L‐NMMA. The effectiveness of the α and β adrenergic antagonists was confirmed by the blockade of the respective adrenergic agonists. We conclude that NO seems to act on the brain of frogs as a hypotensive agent via a mechanism dependent on α, but not β, adrenergic receptors in the periphery.

Supported by FAPESP and CNPq.
Protective role of PPARγ receptors against vascular dysfunction associated with insulin resistance
Authors: El‐Bassossy Hany , Fahmy Ahmed , Abo‐Warda Shaimaa
Published: June 2021

Here we investigated the protective effect of PPARγ receptor stimulation from vascular complications associated with insulin resistance (IR).

IR was induced by fructose (10% in drinking water) and left for 12 weeks. PPARγ was stimulated by either rosiglitazone or pioglitazone (5 or 10 mg/kg respectively) in the last 6 weeks. Then, non invasive blood pressure (BP) was recorded. Rings of isolated thoracic aorta were used to measure the responses to phenylephrine (PE), KCl, acetylcholine (ACh) and sodium nitroprusside (SNP) relaxants and to study NO generation and intracellular calcium.

IR was associated with elevated systolic and diastolic BP while, both rosiglitazone and pioglitazone significantly reduced the elevation in BP. IR increased response of aorta to PE and KCl, decreased response to ACh without affecting response to SNP, while both rosiglitazone and pioglitazone prevented the impairment in response to KCl and PE but not to ACh. IR was accompanied with reduced ACh stimulated NO generation but exaggerated intracellular calcium increase in response to KCl while both rosiglitazone and pioglitazone restore normal KCl stimulated increase in intracellular calcium but did not affect NO generation.

In conclusion, PPARγ stimulation protects from IR induced elevation in BP through preventing impairment in vasoconstriction and calcium movement.
ACTH 22‐39 increases food intake and anxio‐like behaviors in chicks
Authors: Reid Caitlin Ann , Smith Marissa L.
Published: June 2021

The c‐terminal fragment of adrenocorticotropic hormone (ACTH 22‐39) is a well‐known insulin secretagouge primarily associated with increased food intake. The present study investigated the effects of ACTH 22‐39 on food and water intake, ingestive and non‐ingestive behaviors, and glucose concentrations in chicks. In Experiment 1, ACTH 22‐39 increased food intake at all observation times through 180 min post injection but did not affect water intake or whole blood glucose concentration. Although ACTH 22‐39 is an insulin secretagouge, and insulin decreases glucose concentration, ACTH 22‐39 increased food intake which decreases insulin, which may account for the lack of a net effect on glucose concentration. In Experiment 2, anxio‐like behaviors were significantly increased following ACTH 22‐39 indicating a potential influence of ACTH 22‐39 in the stress response. These data may be used to develop future pharmacological treatments for obesity and anorexia.
Induction of non‐cardiogenic pulmonary edema following cerebral hypoxia in a canine model
Authors: Khademi Shadi , Frye Melinda , Jeckel Kimberly , Irwin Dave , Hamilton Karyn , Monnet Eric
Published: June 2021

Neurogenic pulmonary edema (NPE) is a non‐cardiogenic pulmonary edema associated with stroke, traumatic brain injury and seizure with high morbidity and mortality, yet the underlying mechanism(s) is unknown.

Hypothesis

We hypothesize that cerebral hypoxemia induces pulmonary edema in the presence of systemic normoxia.

Methods

Using an updated Moss's model for induction of cerebral hypoxia, 8 Walker hounds (25–30kg) were randomly assigned into cerebral hypoxemia (SaO2 ~55%) with systemic normoxia (SaO2 ~90%) (CH; n=4), cerebral and systemic hypoxemia (SaO2 ~ 50%) (GH; n=2), and cerebral and systemic normoxia (SaO2 ~90%) (Con; n=2). In this model, femoral venous (hypoxic) or arterial (normoxic) perfusate was delivered from a cardiopulmonary bypass circuit to the brain. Outcomes included the wet to dry lung weight ratio as an index of pulmonary edema, hemodynamic measurements, intracranial pressure (ICP) and cerebral blood flow.

Results

Wet to dry weight ratio was increased in the CH and GH groups compared to control. We observed no changes in hemodynamic parameters in CH dogs, with the exception of an increase in ICP. Dogs with GH exhibited increased cardiac output, pulmonary artery pressure and cerebral blood flow, with decreased systemic blood pressures; no increase in ICP was observed.

Conclusion

Brain hypoxemia, in the absence of systemic hypoxemia, induces pulmonary edema in this canine model. DARPA N66001‐10‐C‐2134.
Perinatal hypoxia increases KCNQ channel activity in adult rats
Authors: Sedivy Vojtech , Hnilickova Olga , Smolkova Pavlina , Herget Jan
Published: June 2021

Hypoxic stress at birth is linked with increased susceptibility to various diseases in adulthood. Perinatal hypoxia causes irreversible changes of the pulmonary vascular bed, some of which are more expressed in females. KCNQ (subgroup of voltage gated K+) channels have been suggested to play a role in pulmonary vascular reactivity. We hypothesized that perinatal hypoxia alters KCNQ channel activity in adulthood.

Rats of the perinatal hypoxic group (PH) were born in a normobaric hypoxic chamber (FiO2 0.12; one week before the delivery) and kept in the chamber for one week after birth. Then they were raised in atmospheric air. Normoxic controls (N) were born and kept in room air. Both groups were divided according to gender and studied at the age of 20 weeks. We measured changes in perfusion pressure in isolated, saline‐perfused lungs. Dose‐response to a specific KCNQ channel inhibitor linopiridine (0.5–16 μM) was tested.

In contrast to males, PH females have increased basal perfusion pressure and increased reactivity to K+ than N females. PH rats of both sexes are more sensitive to linopirdine. The increase in perfusion pressure in response to the highest linopiridine dose was 10.2 ± 1.6 mmHg in the PH group and only 5.0 ± 0.8 mmHg in the N group.

Exposure to hypoxia in perinatal period increases KCNQ channel activity in adult rats.

Supported by Cardiovascular Research Center MSMT 1M 0510 and Czech Science Foundation 305/08/0108.
Fat chance: the effect of lipid supplementation on adapting to hypoxia in skeletal muscle cells
Authors: Kanatous Shane B , Sclater Amber E , DeMiranda Michael A , Corley Alyssa E
Published: June 2021

Diving mammals are a unique type of endurance athlete that undergo apnea during exercise, yet rely on an aerobic lipid based metabolism to fuel their primary swimming muscles. A hallmark adaptation that allows diving mammals to endure aerobic exercise in a seemingly hypoxic environment is the high abundance of myoglobin in their working skeletal muscle. Our previous research found in response to hypoxic conditions alone the expression of myoglobin significantly decreased in mouse cell lines. In contrast, recent results showed that unlike terrestrial mammal muscle cells the concentration of myoglobin increased under hypoxic conditions in seal primary muscle cells. The purpose of this study was to determine the effects of lipid supplementation on myoglobin expression in mouse C2C12 and seal primary muscle cells. Myoglobin assays were used to quantify differences in myoglobin expression. As previously seen in our seal muscle cells, myoglobin expression was significantly increased in C2C12 cell lines with lipid supplementation under hypoxic but not normoxic conditions, thus implying that lipids play a role in driving the expression of myoglobin under hypoxic conditions in mammalian muscle cells.
Heat stress‐induced autophagic regulations in rat skeletal muscle
Authors: Yoshihara Toshinori , Naito Hisashi , Kakigi Ryo , Ichinoseki‐Sekine Noriko , Sugiura Takao , Katamoto Shizuo
Published: June 2021

PURPOSE

To examine the effects of acute heat stress on the regulation of autophagy in rat skeletal muscle.

METHODS

Twenty‐eight male Wistar rats (15 wk‐old) were used in this study. After 12h fasting, one leg was immersed in a hot water (43°C) for 30 min under anesthesia. The other leg was served as a control one. During the heat treatment, muscle temperature was recorded by a needle thermometer. The soleus muscle in both legs were removed before (Pre, n=4) and 0 (Post0, n=6), 30 (Post30, n=6), 60 (Post60, n=6) min or 24 hours (Post24h, n=6) after the heat stress. Phospho‐ and total‐Akt and mTOR, and LC3‐I/II protein expressions in the both muscles were determined by Western blotting technique.

RESULTS

Muscle temperature of the heat‐stressed leg was gradually increased to 41.8±0.3°C. In the soleus muscle, the levels of the autophagic makers LC3‐II were significantly increased in the Post60 group compared with Pre, Post30 and Post24h. Also, LC3‐I protein expressions were significantly decreased in the Post24h compared with Post0, Post30 and Post60. The Akt phosphorylation ratios of Post0 were significantly higher than any other groups. Moreover the phosphorylation level of mTOR was significantly increased in Post0 compared to Pre and Post24h.

CONCLUSION

These results show that acute heat stress lead to not only the activations of Akt/mTOR pathway but also the facilitation of protein turnover via autophagy.
Splanchnic, rather than renal nerves contribute to the maintenance of Dahl salt‐sensitive (SS) hypertension
Authors: Foss Jason , Fink Gregory D , Osborn John W
Published: June 2021

The sympathetic nervous system has been implicated in the pathogenesis of salt‐sensitive hypertension; however, the specific sympathetic targets involved remain unclear. The present study tested the hypothesis that splanchnic nerves play a greater role than renal nerves in the maintenance of elevated blood pressure in the Dahl SS rat, a genetic model of salt‐sensitive hypertension. Dahl SS rats were fed a low salt diet (0.1% NaCl) and instrumented with radiotelemeters to measure mean arterial pressure (MAP). After 10 days of recovery and a 4 day baseline period, rats were put on a high salt diet (4% NaCl) for 6 weeks. On average, MAP increased approximately 25 mmHg by the end of the 3rd week of high salt. At this time the rats were subjected to either celiac ganglionectomy (CGX; n = 4), renal denervation (RDNX; n = 3) or a sham surgery (SHAM; n = 4). Immediately following surgery, MAP transiently dropped in all groups and then slowly increased over the remainder of the protocol. This increase, however, was markedly attenuated in CGX rats. By 3 weeks post surgery, the change in MAP from the day before surgery was significantly less in CGX rats (3.0 ± 2.2 mmHg) than in RDNX (14.8 ± 3.2 mmHg) and SHAM (19.5 ± 1.1 mmHg) rats. These data support the hypothesis that the splanchnic, but not renal, nerves contribute to the maintenance of Dahl SS hypertension. Supported by R01HL076312.
Electrophysiological and molecular characterization of the two‐pore‐domain Potassium channels in murine fibroblasts of renal cortex
Authors: Kacik Michael , Kaistha Brajesh , Hoyer Joachim
Published: June 2021

Background

Potassium channels have been proposed to play a key role in the progression of organ fibrosis. Recently, it has been shown that potassium channels control crucial fibroblast functions like proliferation and migration (Grgic., 2009).

Our aims in this study are to characterize the role of new classes of potassium channels in murine renal fibroblasts (TFBs) and determine the regulation of the activity of these newly characterized potassium channels by different growth factors that are important in the pathophysiology of renal fibrosis.

Method

Channel characterization were determined by whole cell patch‐clamp experiments of TFBs. In addition to channel function we tested the expression with real‐time‐PCR and the localization with immunofluorescence experiments.

Results

In murine renal fibroblasts we detected the expression of six members of the two‐pore‐domain potassium channels (K2P) family. In the quantitative RT‐PCR experiments, TREK‐1, TREK‐2 and THIK‐1 were the most dominant arachidonic‐acid (AA) sensitive K2Ps.

In whole cell experiments, we tested activators or inhibitors like changes in pH and temperature, mechanical stress, volatile anaesthetics, unsaturated fatty acids, metallic substances, antiarrhythmic and antidepressive agents. Our data suggest that THIK‐1 has a significant contribute to AA‐sensitive current.

Mitogenic stimulation of fibroblast with profibrotic agents like b‐FGF and TGF‐β led to a downregulation of the channel activity in more than 70 %.

Conclusion

Aside from previously described Ca2+‐activated potassium channels renal fibroblasts also possess a distinct array of K2P‐potassium channels. These channels are also regulated in the presence of profibrotic stimuli. In the clinical context K2P‐channels should be tested as pharmacological target in renal fibrosis.
EPR and UV‐Vis studies of the Fenton reaction with the feasibility of using H2O2 generated by Abeta42 and its interaction on AChE catality activity
Authors: Mendez Armando , Rosales Martha Cecilia , Hernandez Maricarmen , Zamorano Rafael , Ramirez Daniel
Published: June 2021

Two of the most important biomolecules involved with memory and learning are the acetylcholinesterase (AChE) and beta amyloid. The first one hydrolyzes the neurotransmitter acetylcholine (ACh). The second biomolecule is located in the brain forming clusters with glial cells, iron and amyloid fibrils of 42 residues (Abeta42). The production of hydrogen peroxide (H2O2) from Abeta42 fibrils, as well as the possibility that the H2O2 produced in this way it reacts with iron (Fe) via Fenton reaction, producing free radicals hydroxyl (.OH) has been proposed. These reactive oxygen species could have some effect on the AChE activity. In this work enzyme kinetics was studied by UV‐Vis, while free radical generation .OH was followed by electron paramagnetic resonance (EPR). The results show no change in the bovine AChE enzyme activity and that Abeta42 does not produce H2O2.
Involvement of vascular endothelial growth factor (VEGF) secretion in radiation surviving tumor cells derived from breast cancer cell line
Authors: Tsutsumi Kaori , Chiba Ayaka , Ooshima Takahito , Yamazaki Rie , Nishioka Takeshi
Published: June 2021

Radiotherapy is an effective approach for the treatment of many types of cancer. However, it has been reported that the malignant capacities of tumor cells can be increased in those tumors that survive radiotherapy. Here, we constructed radiation‐ surviving tumor cells (IR cells) by using the human breast cancer cell line MCF7, in the manner of our previous report. We focused on angiogenesis in IR cells, because angiogenesis in breast cancer is highly associated with unfavorable patient prognosistic criteria, such as tumor metastasis and relapse. To analyze angiogenesis in IR cells, we performed a tube formation assay and plasma clot assay by using human umbilical vein endothelial cells (HUVEC). However, IR cells had the same levels of angiogenic activity compared to that of parental cells. On the other hand, the secretion of vascular endothelial growth factor (VEGF) was enhanced by about 1.9‐fold in IR cells as estimated by ELISA. From microarray analysis, the gene expression of NRP1 and CXCL14 was upregulated in IR cells. These results suggest that increased CXCL14 expression prevents angiogenesis in IR cells, even though NRP1 may promote upregulation of VEGF expression by autocrine signaling. Further studies are necessary to clarify the function of upregulated‐VEGF in IR cells, and may elucidate their possible role in controlling the behavior of tumors surviving radiotherapy.
Catalytic transitions in the MDR1 P‐glycoprotein probed by target molecular dynamics
Authors: Wise John G
Published: June 2021

A serious problem for cancer and viral chemotherapies is the active efflux of drugs catalyzed by ABC‐transporters. Human P‐glycoprotein (Pgp), one such problematic transporter, is a transmembrane protein composed of two nucleotide binding domains (NBD) and two transmembrane domains (TMD) in a NBD1‐TMD1‐NBD2‐TMD2 topology. Two ATP binding sites are created from both NBDs and drug binding sites (DBS) are created from both TMDs. It is thought that binding of ATP engages the two NBDs and hydrolysis leads to a repositioning of the DBS from inward facing to outward facing conformations, but the details of these complex conformational changes remain unknown. Targeted molecular dynamics (MD) techniques enable the sampling of large conformational changes in a protein that would normally be inaccessible due to large energy barriers. In this work, targeted MD of Pgp using 4 target structures was performed starting with fully disengaged NBDs and inward facing DBS and ending with fully engaged NBDs with fully opened outward DBS. 26 nonredundant transitional protein structures were identified by statistical methods and drug docking of 23 drugs was tested on each structure. Previous biochemical studies strongly support the results obtained here. The combination of protein dynamics and dynamic ligand docking used here revealed new details of the Pgp transport mechanism. Supported by NIH 1R15GM094771‐01A1 to P.D. Vogel and JGW.
Expressioneering™ Technology Accelerates Functional Expression and Crystallization of GPCRs for Drug Discovery
Authors: Sen Saurabh , Franz Laura , Mead David , Steinmetz Eric
Published: June 2021

Structural biology of membrane proteins and GPCRs has seen major advances over the past few years, but overcoming the challenges of functional GPCR expression and structural stabilization are still time‐consuming and expensive. Our approach combines a powerful cloning strategy with novel transmembrane guides and co‐expressed helper proteins to significantly increase functional receptor expression and membrane insertion in E. coli.

Lucigen's Expressioneering™ technology is a rapid and simple recombination based expression cloning method. We present here the use of this technology to optimize GPCR expression through fusion to a variety of transmembrane guides and novel visualization tags. We have been successful in obtaining conformationally active functional GPCRs with this system. We will also discuss unique new strategies for conformational stabilization of GPCRs through use of novel fusion partners and interacting proteins. Preliminary data using a novel fluorescent protein in the IL3 demonstrates retention of functional activity and may promote GPCR crystallization. Data obtained using Lucigen's Expresso CMV system are comparable with those obtained from the bacterial system.
Characterization of an anti‐pathogenic lactonase from Palk Bay metagenome
Authors: PANDIAN SHUNMUGIAH THEVAR KARUTHA , NITHYA CHARI , VISZWAPRIYA DHARMAPRAKASH
Published: June 2021

Objectives

Screening of metaclones for anti‐pathogenic activity and identification of anti‐pathogenic agents.

Methods

Screening for anti‐pathogenic activity using Chromobacterium violaceum (ATCC 12472) and CV026 as indicators. Mode of action by TLC (Thin Layer Chromatography), HPLC (High Performance Liquid Chromatography) and PCR (Polymerase Chain Reaction).

Results

Recombinant fosmid clones (N=10200) were propagated in E. coli DH10B. When individually 1056 clones were used to challenge C. violaceum CV026, eight clones could able to inactivate the perception of QS signal HHL (C6‐HSL) by the Chromobacterium biosensor. Additional assays revealed that the metagenomic clone M8‐70 disrupted AHL signals, including C6‐HSL, heptanoyl‐ (C7‐ ), octanoyl‐ (C8‐) and decanoyl‐homoserine lactone (C10‐HSL), as well as the hydroxy‐ or oxo‐substituted derivatives at carbon 3, such as OC6‐HSL, OC8‐HSL and OHC8‐HSL. Culture supernatants of M8‐70 was examined for its effect on P. aeruginosa PAO1 (LasA, LasB, Pyocyanin, Pyoverdin and and Total protease), S. marcescens (prodigiosin production, swarming, secreted caseinase and lipase) QS system and distinct profiles were obtained for las and rhl QS genes. Furthermore, the growth curve analysis and cell count method made it clear that the supernatant of M8‐70 had no antibacterial activity toward P. aeruginosa PAO1 and S. marcescens. The results of the ring closure assay and the sequence analysis of the PCR products from M8‐70 (GenBank accession number HQ876560) revealed that its activity as lactonase activity.

Conclusion

In this study a lactonase is reported through metagenomics approach from the sediment of Palk Bay which was found responsible for the inhibition of QS mediated virulence factors of Gram negative bacterial pathogens without inhibiting their growth.
Mediterranean Diet adherence is decreasing in healthy subjects over the years 1996–2010 also in Mediterranean area.
Authors: Trovato Guglielmo M , Martines G. F. , Catalano D.
Published: June 2021

Mediterranean Diet score (range 0–55) was assessed in 8138 healthy non‐diabetic, overweight/obese subjects, 1996–2010, referred for US liver diagnostic and dietary counseling. Mediterranean food (Pasta and rice;whole‐grain bread, brown rice, legumes; Fruit; Green vegetables; Fish, poultry; Nonfat or low‐fat dairy products; olive oil) had assigned a score of 0: no consumption, a score of 1 for 1 to 4 times/w, 2 for 5 to 8 times/w, 3 for 9 to 11 times/w, 4 for 12 to 14 times/w, and 5 for more than 14 times/w; for westernized food (Red meat; Dairy products‐butter; Potatoes and eggs; Cakes) opposite scores were assigned (i.e., 0 when a participant reported more than 5 weekly consumption to: 5 for no weekly consumption, score 4 for 1 weekly consumption, 3 for 2 weekly consumption, 2 for 4 weekly consumption, 1 for 5 weekly consumption); wine and alcoholics (0–10 g of alcoholics from Red Wine for women score 5; 0–20 g of alcoholics from Red Wine for men score 5; each increment of 10 determines negative scores (20–30= −1. 30–40=−2. 40–50=−3. 50–60=−4, >60= −5 for men; 10 less for women and for all non‐wine alcoholics,: 10–20= −1; 20–30= −2; 30–40= −3; 40–50= −4; >50 = −5).

There is a trend toward the loss of the adherence to Mediterranean Diet from 37.06±3.213 to 34.82±5.014 p<0,0001, not associated with BMI or physical activity change.

Adherence to Mediterranean Diet is significantly yearly decreasing in Italy in healthy subjects.
Autoimmune disease (AID) in anorexia nervosa and bulimia nervosa
Authors: Gerhardt Ann
Published: June 2021

We explored an impression, gained from a general health status questionnaire, that AID prevalence may be high in eating disorder patients. In the original study, 79 of 488 former eating disorder patients of an internal medicine practice, age ≥30 years at the time of the study, were found and 73 had completed a general health questionnaire with 30 questions related to 10 organ systems. No question mentioned any specific autoimmune disease. We contacted patients for more information about diagnoses.

Results

Fifteen women (20.5%) had one or more AID diagnosis. These included Hashimoto's thyroiditis, celiac disease, systemic lupus erythematosis, rheumatoid arthritis, Crohn's disease, uncharacterized connective tissue disorder, combined variable immunodeficiency disease, interstitial cystitis, membranous glomerulonephritis and multiple sclerosis. AIDs were more common with advanced age (mean age 47.13± SD 9.27 vs. 41.40± SD 8.53, p=0.026). AID prevalence was significantly more common (34.8% vs. 14%, p=.041) in women who abused laxatives (N=23).

Conclusion

AIDs are common in this eating disorder population, and may be related to laxative abuse. The 20.5% prevalence is high compared to an estimated AID prevalence of 14% in all women ≥ 30 years of age, especially since none had Type I diabetes mellitus or psoriasis, the most common AIDs.

Support from Sutter Institute for Medical Research
Lessons Learned in Conducting Nutrition‐Related Research in African American (AA) Older Adults
Authors: BABATUNDE OYINLOLA TOYIN , CAMPA ADRIANA , HIMBURG SUSAN P
Published: June 2021

Background

To reduce health disparity, the implementation of culturally sensitive strategies to improve participation of minority groups in nutrition research is needed. The objective of this report is to present lessons learned from outreaching and recruiting community‐based minority elders.

Methods

Participants were recruited through gatekeepers who were trusted by the target population and who belonged to the Churches and Senior Centers where African American (AA) elders routinely attend in three South Florida counties. Accessing and recruiting older AAs was challenging without these gatekeepers who acted as liaison between the church, group leaders, the participants and the investigator to negotiate agreement to participate in research. All data‐collecting forms were field‐tested to adapt them culturally. After 2 years, 130 participants were recruited, consented and participated in a six‐week education sessions on osteoporosis prevention.

Results

Mean age of participants was 70.1 ±SD 10.4 y., 90% were women, and median education was 13 y. (range 8–20 y.). Attrition rate was 15.4%, 110 (84.6%) completed the program.

Lessons Learned

Accessing AA older adults, an underserved and hard‐to‐reach group, requires determination and perseverance. Trusted gatekeepers, cultural adaptations and familiar social settings are needed to improve participation of minority groups in nutrition‐related research to reduce health disparity.
Head Start Preschool Teachers’ Willingness to Implement Nutrition and Agricultural‐Based Learning Centers in the Classroom
Authors: Dunham Christopher E , Cash Rebekah , Carraway-Stage Virginia , Goodell L Suzanne
Published: June 2021

A learning center is an educational station where preschool students “make a choice” during class time. These centers allow children to utilize self‐paced learning to develop sensory, fine‐motor, and critical thinking skills. Learning centers are a familiar part of the preschool classroom; however, the motivations for implementing certain centers as well as the overall efficacy that centers provide are rarely studied. Researchers designed centers to use nutrition and agriculture‐based topics to teach concepts in science, manipulatives, language arts, dramatic play, and art. Specific educational topics included concepts such as nutrition of fruits and vegetables, planting, and composting. For example, “Bubbly Bread” in the science center engaged children in an experiment with yeast and sugar to learn about fermentation through observation. An attitudinal survey assessed teacher attitudes towards learning centers that emphasized fruits and vegetables as a method to incorporate nutrition education. Formative assessment was used to determine teacher willingness to repeat each observed learning center within their classroom, ability to obtain materials used in each learning center, and any related barriers that would prevent future implementation. Results from this study are being used to enhance the development of nutrition and agricultural‐based educational materials for preschool children.
Knowledge, Awareness of Diabetes Mellitus, and Weight Change Among Iranian Adults Living in the United States
Authors: Etemadinejad Haleh , Kirk Elizabeth , Kloubec June , Koutoubi Samer
Published: June 2021

The purpose of this study was to measure knowledge and awareness of diabetes mellitus and weight change among Iranian adults living in the United States (U.S.). In a cross‐sectional design, a recruitment flyer was used at Iranian grocery stores, and other public places where Iranian‐Americans congregate. Interested subjects received a link to access and complete an online survey. Questionnaires were completed by 171 Iranian adults living in the greater Seattle, WA area. Questionnaires included personal characteristics, weight change, and knowledge and awareness of diabetes. “Knowledge” was defined as: knowledge of risk factors, symptoms and complication, treatment and management, and monitoring. Among subjects, 76.0% reported weight change of more than five pounds since migration to the U.S. (p<0.001), compared to those without any weight change. Overall, the mean of weight gained for the subjects was 17 ± 7lbs. Mean knowledge of diabetes was 74% ± 2 (on a scale of 0–100). Mean knowledge of risk factors for diabetes was 50% ± 1. Mean knowledge of symptoms and complications was 86% ± 2. Mean knowledge of treatment and management was 73% ± 3. Mean knowledge of monitoring was 85% ± 1. These findings suggest that Iranian adults living in the U.S. demonstrated a high knowledge of diabetes. In addition, many adult Iranians gained weight since migrating to the U.S.
Genetic polymorphisms in AHR and CYP1A2 are associated with habitual caffeine consumption and are modified by age and smoking
Authors: Josse Andrea R , Costa Laura A Da , Campos Hannia , El-Sohemy Ahmed
Published: June 2021

Recent genome‐wide association studies (GWAS) from populations of European decent have identified polymorphisms in AHR and CYP1A2 that are associated with caffeine/coffee intake. We examined whether these polymorphisms in AHR (rs6968865 and rs4410790) and CYP1A2 (rs2472297 and rs2470893) were associated with caffeine consumption in a Costa Rican population. Subjects (n=4,612) were participants from a case‐control study of gene‐diet interactions and myocardial infarction. Those with hypertension (n=1,692), or missing information on genotype (n=389), caffeine intake (n=4) or smoking status (n=4) were excluded. Sequenom MassARRAY was used for genotyping, and caffeine intake was assessed by a validated food frequency questionnaire. Compared to those consuming <100 mg/d of caffeine, subjects consuming >400 mg/d were more likely to be carriers of the T, C or T allele for rs6968865, rs4410790 and rs2472297, respectively. The corresponding odds ratios (OR [95% CI]) were 1.41 (1.03–1.93), 1.41 (1.04–1.92) and 1.55 (1.01–2.36). The multivariate‐adjusted ORs (95% CI) for rs6968865 were 1.44 (1.03–2.00) for all subjects, 1.75 (1.16–2.65) for non‐smokers and 1.15 (0.58–2.30) for smokers, and 2.42 (1.45–4.04) for >57 yrs and 1.00 (0.65–1.56) for ≤57 yrs. Our findings confirm previous results from GWAS linking polymorphisms in AHR and CYP1A2 with caffeine consumption, but only in non‐smokers and older adults.

Grant Funding Source: Canadian Institutes of Health Research (CIHR)
Constitutive activation of mTOR pathway by leucine causes heart hypertrophy which can be blocked by rapamycin
Authors: Davoodi Jamshid , Hutson Susan
Published: June 2021

Mitochondrial branched chain aminotransferase (BCATm) KO mice fed an amino acid diet containing intermediate levels of branched chain amino acids (BCAA) to prevent toxicity and wild type mice fed an amino acid diet with normal levels of the BCAA were studied. To determine the mTOR specific effects of constitutively high concentration of BCAA in the blood, BCATm KO and wild type mice were fed diets with and without encapsulated rapamycin. Following thirteen days of feeding, the mice were sacrificed without fasting. The plasma levels of BCAAs were found to be higher in the KO mice without any effect by rapamycin. Despite higher concentration of leucine, food intake was higher for the KO mice suggesting that chronically high leucine levels cannot suppress food intake. BCATm KO mice exhibited heart but not skeletal muscle hypertrophy which was prevented by dietary rapamycin. In addition, feeding the KO mice with a diet containing rapamycin resulted in increased activity. Rapamycin feeding had no effect on heart size of the wild type mice although mouse activity was reduced. Western blotting showed the activation of mTOR signaling pathway in KO mice. Our results suggest that leucine induced protein synthesis through the activation of mTOR that leads to growth is tissue dependent and leucine alone is insufficient to induce skeletal muscle hypertrophy. (Support Ajinomoto and Virginia Tech)
Estimating the RBC omega‐3 index and breast milk DHA from dried samples collected on filter paper
Authors: Harris William Stephen
Published: June 2021

Assessment of the omega‐3 fatty acid status of both human blood and breast milk is important in studies that examine the relationships between these fatty acid levels and health outcomes. Both blood and milk samples are difficult to collect and process in the field, hence improved approaches to sample acquisition are needed to expand the research base for these metrics. We have therefore developed methods to collect, preserve, transport and analyze the fatty acid composition of dried blood spots (DBS) and dried milk spots (DMS). For both sample types, a single drop of blood/milk were applied to Whatman 903 cards which had been pretreated with an antioxidant preservative cocktail (OxyStop®). Frozen human milk samples (n=5) were thawed and analyzed immediately or spotted on cards and placed in a drawer at room temperature for 5 days. The DHA content of the liquid vs. dried milk samples were analyzed in triplicate. Mean DHA levels were 0.19% and 0.20%, respectively, with an r2 = 0.99 (p<0.01). RBC EPA+DHA (the omega‐3 index) was estimated from DBSs vs. direct RBC analysis in 106 healthy subjects. The correlation was r2 = 0.92 (p<0.0001). Thus both RBC and milk omega‐3 content can be accurately assessed from samples collected and transported on filter paper. These studies were funded with internal support.
Association between the plasma proteome and plasma alphatocopherol concentrations in humans
Authors: Da Costa Laura Anne , García-Bailo Bibiana , Borchers Christoph H , Badawi Alaa , El-Sohemy Ahmed
Published: June 2021

Vitamin E has been inversely associated with chronic disease; however, the biological processes regulated by this vitamin have not been fully elucidated. The objective of the present study was to examine the association between α‐tocopherol and the plasma proteome. Subjects are from the Toronto Nutrigenomics and Health Study and include men and women (n=1,022) who completed a general health and lifestyle questionnaire and 196‐ item food frequency questionnaire and provided a fasting blood sample. Levels of α‐tocopherol were analyzed using a reversed‐phase isocratic HPLC method with fluorescent detection. Fifty‐four plasma proteins were assayed by a mass spectrometry‐based multiple reaction monitoring method. Analysis of covariance was used to compare mean concentrations of plasma proteins across tertiles of α‐tocopherol. Levels of apolipoprotein C‐III, fibrinogen alpha, beta, and gamma chains, fibronectin and fibrinopeptide A were significantly and positively associated to plasma α‐tocopherol, while intermediate levels of α‐tocopherol were significantly associated with higher levels of alpha‐1B‐glycoprotein (all P<0.0009). In conclusion, circulating levels of α‐tocopherol were significantly associated with plasma proteins from several physiological pathways. This research was funded by the Advanced Foods and Materials Network.
Vitamin C Status is associated with Physical Fitness and Activity Indices in College Men: a cross‐sectional study
Authors: Johnston Carol S , Smith Lisa L , Schlueter Amanda K , Swan Pamela D
Published: June 2021

Fruit and vegetable consumption has been shown to predict physical activity (PA) in college students. This study examined the specific association between plasma vitamin C, found predominately in fruits and vegetables, and indices of PA in college students. Fifty healthy, non‐smoking men who did not take vitamin supplements or engage in regular vigorous activity were recruited. Participants (23.2±0.6 y; 24.7±0.5 kg/m2) provided a fasting blood sample and a 24‐h recall, and completed PA assessments including a step test. Participants were fitted with a pedometer and recorded steps for three complete days. Vitamin C concentrations ranged from 12 to 50 μmol/L. Diet quality was not correlated with plasma or dietary vitamin C. Participants with inadequate vitamin C status (<28 μmol/L, n=21) had significantly lower fitness levels and daily step counts, and were less likely to engage in moderate‐to‐vigorous exercise compared to vitamin C‐adequate peers. These relationships between vitamin C status and PA were independent of diet quality and fruit and vegetable consumption. These data indicate that college men are at risk for marginal vitamin C status, a condition linked to reduced fitness and PA.
Effect of paternal folate deficiency on brain development in postnatal rat pups
Authors: Kim Hye Won , Kim Ki Nam , Choi Yun Jung , Lim Eunmi , Mejos Karen Kay , Lee Hwayoung , Chang Namsoo
Published: June 2021

This study was performed to evaluate the effects of combining paternal folate sufficiency (PS) or deficiency (PD) with maternal folate sufficiency (MS) or deficiency (MD) on histological and electron microscopic observations in the brain of pups. Experimental diets of different folic acid content (0 vs. 8 mg/kg diet) were fed for 4 weeks prior to mating and pups were killed on day 21 of lactation. The expression of brain‐derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP) were analyzed by immunohistochemistry, and ultrastructural changes of hippocampal neuronal cells were evaluated by electron microscopy. As compared to the PS×MS group, the hippocampal BDNF expression was decreased and GFAP expression was increased in the PS×MD, PD×MS and PD×MD groups. The hippocampal MBP expression was however, decreased only in the PD×MD group. In the cortex, there were no differences among the four groups. In the PS×MD, PD×MS and PD×MD groups, neurons with segmented or fragmented nuclear chromatin, irregular invagination of the nuclear membrane into the nucleoplasm and intranuclear vacuolations were observed, meaning apoptotic cell death. These results suggest that the impact of paternal folate status might be as important to the normal growth and development of the brains of pups as maternal folate status.
Serum 25(OH)D, 1,25(OH)2D and parathyriod hormone responses to vitamin D supplementation in early pubertal children: a dose‐response trial
Authors: Lewis Richard , Laing Emma , Weaver Connie , Peacock Munro , Hausman Dorothy , Hall Daniel , Martin Berdine , McCabe George , Warden Stuart , Hill Kathleen
Published: June 2021

Serum 25(OH)D, 1,25(OH)2D and parathyroid hormone (PTH) responses to vitamin D3 supplementation in a dose‐response manner in children are unknown. Dose‐response curves for vitamin D metabolites were generated in 320 male and female, black and white children in the early stages of puberty (9–13 y) who participated in a multi‐site, randomized, triple‐masked trial, received placebo or vitamin D3 [400, 1000, 2000 and 4000 IU/d (n=64/group)] and were sampled 5 times over 12 wks. The intervention took place in the winter months at two US latitudes, 34°N and 40°N. Serum 25(OH)D and 1,25(OH)2D were assessed using RIA and PTH, by IRMA. Asymptotic and linear regression models were fit for vitamin D metabolites and PTH, respectively. Baseline 25(OH)D and 1,25(OH)2D values were similar between doses. Increases in 25(OH)D from baseline to asymptote depended on dose and differed between blacks and whites, with larger gains for whites at the 4,000 IU dose (p<.05). Serum 1,25(OH)2D exhibited an asymptotic pattern of increase over time, with the increase dependent on dose (p=.07), but did not differ across races. PTH data showed no significant dependence on dose. Vitamin D supplementation with 400 IU maintains wintertime 25(OH)D values in children. Whether the differential 25(OH)D response to 4000 IU in whites and blacks has health implications deserves further inquiry. Supported by NICHD, R01HD057126.
Anti‐osteoporosis effect of Essential Oil from Plant Z‐1
Authors: Kim Gun-Hee , Chang Kyung Mi
Published: June 2021

The volatile aroma constituents of plant Z‐1. were separated by the steam distillation extraction (SDE) method using a Clevenger‐type apparatus, and analyzed by gas chromatography‐mass spectrometry (GC/MS). Limonene (18.04%), geranyl acetate (15.33%), and cryptone (8.52%) were the major volatile flavor compounds of plant Z‐1. The effects of plant Z‐1. essential oil (EO) from on the function of osteoblastic MC3T3‐E1 cells were tested. It increased the collagen and mineralization of osteoblasts (P<0.05), indicating that plant Z‐1. EO may help prevent osteoporosis.
Investigating piglet kidney vitamin A concentrations after high dose supplementation at birth
Authors: Gannon Bryan M , Arscott Sara A , Tanumihardjo Sherry A
Published: June 2021

Potential metabolism and toxicity issues with infant vitamin A dosing procedures have led to a vested interest by the World Health Organization. Whole body retinoid trafficking was under investigation by determination of serum, liver, lung, kidney, adrenal, and spleen retinoid concentrations of piglets born from vitamin A depleted sows and dosed at birth with 25K, 50K, or 200K IU preformed vitamin A. These doses represent levels currently administered to newborn children and mothers in regions with high prevalence of vitamin A deficiency. Retinoids are known to be stored in these tissues with kidney being the primary extrahepatic storage site. In this study, kidneys were analyzed by grinding with sodium sulfate and adding C‐23 alcohol as an internal standard. Dichloromethane was used to extract retinol and retinyl esters. All samples were saponified, extracted, and injected onto an HPLC system to analyze total retinol content. A subgroup was also analyzed unsaponified to elucidate the retinyl ester profile. Data will be used collectively with other organ and serum data to have a complete model for vitamin A circulation and storage under a deficient status followed by high dose paradigm.

Grant Funding Source: World Health Organization
Protective Effects of Combined Treatment of Green Tea and Exercise in the Mouse AOM/DSS Colitis‐associated Carcinoma Model
Authors: Kim Si-Young , Lee Young-Soo
Published: June 2021

This study was aimed at investigating the possible protective effects of combined pre‐treatments of green tea and exercise (GE) in the AOM‐initiated and DSS‐promoted (AOM‐DSS) mouse colitis‐associated carcinoma model. Male ICR mice (5 weeks old, n=48) were divided into 8 groups (6 weeks or 12 weeks ‐ control, AOM‐DSS, GE, GE and AOM‐DSS). Green tee (50 mg/kg body weight/day) suspended in 0.5% CMC was given by gavage (4 times/week). The control mice received only 0.5% CMC. The exercise sessions consisted of running with the treadmill speed set at 70% of each animal's maximum velocity (60 min/day, 4 times/week). The mice were fed a diet supplemented with 2.5% DSS for 7 days after AOM initiation in carcinogenesis groups. Our results showed that AOM‐DSS mice with 6‐week and/or 12‐week pre‐treated with GE had lower tumor incidence, lower colon tumor multiplicities and smaller polyps compared with each control group. 6‐week and/or 12‐week pre‐treatment of GE suppressed the expression of MDA and 4‐HNE, and iNOS after AOM‐DSS as well as increased induction of Nrf2 and phase 2 detoxifying enzymes, including HO‐1 and NQO1. These findings suggest that combined pre‐treatments of green tea and exercise may have protective effects against colon cancer by decreasing lipid peroxidation, reducing inflammatory response and increasing antioxidative defense signaling in mice with AOM‐DSS.

Grant Funding Source : Korean Ginseng Corporation
Effect of fish oil supplementation on bone turnover markers in women on aromatase inhibitors: a pilot study
Authors: Hutchins-Wiese Heather Lea , Feinn Richard , Kerstetter Jane E , Tannenbaum Susan , Claffey Kevin , Kenny Anne M
Published: June 2021

Aromatase inhibitors (AI) are adjunctive therapies for estrogen‐receptor positive breast cancer (BC); however, they exacerbate bone loss. We tested the effects of a non‐pharmacological treatment, fish oil, on bone measures in 37 postmenopausal BC survivors on AI. In addition to calcium (1,000 mg/d) and cholecalciferol (800 IU/d), women were randomly assigned to 4 g eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)/d via fish oil or placebo (safflower oil) capsules. Bone mineral density was low in 41% of women; none were osteoporotic. Average age was 61.8±9.7 and BMI 28.0±5.8. There were no between group differences in dietary intake of calories, protein, calcium, vitamin D, n‐3 or n‐6 fatty acids, or other nutrients. Resorption and formation markers (serum C‐terminal telopeptide [sCTX] and procollagen type 1 N‐terminal propeptide [P1NP], respectively) were measured at baseline and 3 months. Baseline sCTX and P1NP were similar between groups. sCTX was reduced by −7.8±20.8% and P1NP by −10.2±16.5% in the EPA+DHA group. The reductions in turnover markers in the EPA+DHA group were not statistically different compared to changes in placebo (sCTX 0.4±20.4; P1NP −3.3±15.9). In this high risk group, nutritional supplementation was not enough to overcome the effects of AI on bone turnover. Project supported by the CT Breast Health Initiative and University of Connecticut Health Center Clinical Research Center.
Anti‐adipogenic Effect of Buckwheat Sprouts treated with Methyl Jasmonate (MeJA) in 3T3‐L1 adipocytes
Authors: Lee Ok-Hwan , Yoon Bo-ra , Lee Young-Jun , Lim Jeong-Ho , Kim Hyun-Jin , Park Kee-Jai , Kim Bum-Keun , Jeong Jin-Woong
Published: June 2021

Methyl jasmonate (MeJA), one of the plant stress inducers, play important roles in the regulation of plant growth and the biosynthesis of the secondary metabolites. Buckwheat sprouts contain various bioactive compounds including rutin (quercetin‐3‐ O‐rutinoside) which have a number of biological activities. We have previously shown that buckwheat sprouts treated with MeJA (TBWE) treatment significantly increased the phenolic compound amounts and the antioxidant activity. The present study is to investigate the inhibitory effects of TBWE on lipid accumulation and ROS production during adipogenesis in 3T3‐L1 cells by analyzing the mRNA expression level of adipogenic transcription factors and pro‐oxidant enzyme. 3T3‐L1 preadipocytes were treated with TBWE for up to 8 days following standard induction of differentiation. During differentiation of 3T3‐L1 preadipocytes, TBWE markedly inhibited lipid accumulation and reactive oxygen species (ROS) production compared with NBWE treated adipocyte cells. TBWE significantly inhibited the mRNA abundance of adipogenic transcription factors (C/EBPα and aP2), pro‐oxidant enzyme (NADPH oxidase, NOX4) and NADPH producing enzyme (glucose‐6‐phosphate dehydrogenase, G6PDH). These results suggest that MeJA significantly increased the amount of phenolic compound, resulting in suppression of adipogenesis and ROS production in 3T3‐L1 cells.