bluuebunny/crossref_metadata_2025 huggingface dataset:
Vitamin D Blocks Estrogen Signaling through PKA
Authors: Ankeny Amanda P. , Schmitt John M.
Published: June 2021

The hormone estrogen (E2) activates a CaM KK, CaM KI, and ERK pathway to promote proliferation of MCF‐7 breast cancer cells. ERK activation is inhibited by cAMP and PKA in certain cells. PKA has numerous cellular substrates including CREB and CaM KK. PKA phosphorylation of CaM KK inhibits its activity and promotes its association with 14‐3‐3γ. Therefore hormones that stimulate PKA activation may in turn block activation of CaM KK and ERK as well as cell proliferation. Our goal was to evaluate PKA's ability to phosphorylate CaM KK and promote its subsequent binding to 14‐3‐3γ in Vitamin D‐treated cells. In addition we examined whether Vitamin D's inhibition of E2‐stimulated ERK activation in MCF‐7 cells utilized PKA using the inhibitor, H‐89. Our results suggest that activation of PKA with Vitamin D inhibits E2 stimulation of ERK activation and increases CaM KK phosphorylation. Vitamin D treatment of MCF‐7 cells, but not E2, increased CaM KK association with endogenous 14‐3‐ 3. Interestingly, Vitamin D induced phosphorylation of CaM KK and ERK inhibition were both reversed by H‐89. Our results suggest that Vitamin D activation of PKA triggers the subsequent phosphorylation and inhibition of CaM KK through its association with 14‐3‐3 preventing CaM KK from activating ERK in MCF‐7 cells.
Evaluating Transgenic Xenopus as a Model System for the Expression of Secreted Proteins
Authors: Laird Joseph G , Marshall Katelyn R , Dean Maria A , Baker Sheila A
Published: June 2021

Model systems such as E. coli, yeast, and Xenopus oocytes are frequently used to express and purify heterologous proteins for subsequent biochemical analysis. However, some vertebrate proteins, particularly those that are extensively modified, can be toxic and very difficult to purify from these systems. In this study we used GFP as a reporter to investigate the feasibility of expressing heterologous proteins from secretory tissues of transgenic X. laevis. Transgenic tadpoles were generated by restriction enzyme mediated integration. GFP was cloned under the control of the larval keratin promoter to drive expression in the epidermis and under the control of the XAG‐1 promoter to drive expression in the cement and hatching glands. GFP expression in the appropriate tissues was observed using both promoters. However, the XAG‐1 promoter caused GFP to be expressed much more uniformly and robustly. Next, we cloned a signal sequence onto GFP so that it would be secreted. The secreted GFP was assayed using Western blots and a GFP fluorescence assay. Secreted GFP was readily collected from the buffer housing either set of transgenic animals but we obtained the highest yields using animals secreting GFP from the cement and hatching glands. In conclusion, expressing proteins specifically in secretory tissues of transgenic tadpoles is a viable method to obtain heterologous proteins for subsequent biochemical analysis.
S. cerevisiae ENV9, ENV10 and ENV11 Are Novel Genes Involved in Lysosomal Vacuole Events and their Products Localize to Lipid Droplets, ER, and Nucleus, Respectively
Authors: Remington Lisa Ann Kiyomi , Hseuh Teli , Gharakhanian Editte
Published: June 2021

ENV9, ENV10 and ENV11 are novel genes uncovered by a genome wide immunodetection screen for mutants defective in processing of the vacuolar enzyme carboxypeptidase Y at late endosome to vacuole interface. Additionally, env9Δ has minor growth sensitivity to caffeine and vesiculated vacuoles, while env10Δ and env11Δ are cold sensitive and the latter has increased fragmented vacuoles. Here, we report on the localization of their GFP and hemagglutinin (HA) tagged constructs as assessed by confocal microscopy and biochemical subcellular fractionation. Env9p localizes to lipid droplets and its possible role in lipid droplet morphology is under investigation. Env10p localizes to the ER and putative negative genetic interactions between ENV9 and ENV10 are under investigation. Env11p localizes to the nucleus and bioinformatic analyses identify a putative BED zinc finger DNA binding domain. Recently, Env11p and Vid22p, another gene product involved in vacuolar protein trafficking, have been reported to colocalize with the transcription factor Tbf1 at selected promoter regions. These findings suggest that Env9p and Env10p play a direct role within the endomembrane flux in vacuolar protein trafficking while Env11p may be indirectly related through transcriptional regulation of vacuolar related genes. NIH‐AREA grant GM085794, CSULB Leonard Scholarship (LR) and an NSF‐MRI grant for the confocal microscope.
Generation of an IgG Monoclonal Antibody to LacdiNAc (LDN) Glycan Epitopes from Splenocytes of Schistosoma mansoni Infected Swiss Webster Mice
Authors: Bartlette Victoria H , Mamphey Bernard , Mandalasi Msano , Nyame Kwame
Published: June 2021

Parasitic helminths of the genus Schistosoma synthesize an assortment of glycoconjugates with unique glycan structures that play immuno‐modulatory role(s) in host‐parasite interactions. The unique feature of schistosome glycans is the presence of LacdiNAc (LDN; GalNAcâ1‐4GlcNAC‐) outer branches instead of the lactosamine (LN; Galâ1‐4GlcNAC‐) branches associated with mammalian glycoconjugates. IgM and IgG antibodies are generated to LDN epitopes during course of schistosome infections in vertebrates. We report the use of splenocytes from Schistosoma mansoni infected Swiss Webster mice to generate IgG monoclonal antibodies to LDN epitope to facilitate the study of the role of LDN antigen in host‐schistosome interactions. Splenocytes were prepared from spleens harvested from Swiss Webster mice at wk 10 post‐infection and fused with myeloma cells to generate hybridomas. The secreted monoclonal antibody Y1H5 was purified over columns of MEP‐hypercel and its specificity for LDN epitopes was analyzed on the glycan array of the Consortium for Functional Glycomics against 660 different glycan structures. The availability of anti‐LDN monoclonal antibody would allow the purification and localization of LDN epitopes during parasite development in intermediate and mammalian hosts.
The role of histidines in neurosteroid binding of NMDA NR2B and D subunits
Authors: Rhoads Sarah , Biello Shannon , Zemcik Laura , Gentile Lisa
Published: June 2021

Implicit in cellular mechanisms essential for learning and memory, NMDA receptors are members of the family of ionotropic receptors. Activation of these receptors through the binding of glutamate results in the opening of ion channels, triggering an excitatory pathway which can be moderated by certain endogenous and exogenous compounds. Previous research with the endogenous neurosteroids pregnenolone sulfate (PS) and 3‐α‐hydroxy‐5‐β‐20‐ one sulfate (PregS) have indicated that the binding of these neurosteroids to NMDA receptors containing the NR2C and NR2D isoforms of the NR2 subunit decreases current flow through NMDA receptors. Binding of PS to NMDA receptors containing NR2A or NR2B subunits causes an increased amount of current flow through the channel, whereas binding of PregS to these subunits decreases the current flow through these channel. Binding studies from our lab have determined that the neurosteroids PS and PregS bind to the amino terminal domain (ATD) of both NR2B and NR2D subunits. Binding of both neurosteroids was monitored between pH 6.0 and 8.0 in 0.5 pH increments. Results demonstrated that the binding of PS to NR2B and D ATDS occurred only between pH 6 and 6.5. PregS showed a similar pH dependence for the NR2B ATD, although binding was restored at pH 8. However, binding of PregS to the NR2D ATD occurred over a wider range, from pH 6–8 with the exception of pH 7.5. The pH dependence of neurosteroid binding suggests the importance of a protonated histidine residue in each binding interaction. The NR2B ATD domain (residues 121–292) contains one histidine (residue 127), whereas the similar NR2D ATD domain contains 3 histidines (residues 143, 182, 202). Results from His to Ala and Gln mutations and chemical modification studies will be presented.
Understanding the regulation of AMPA and NMDA iGluRs through the binding of sulfated neurosteroids
Authors: Bartle Emily Irene , Fanelli David , Roark Ryan , Young Amber , Gentile Lisa
Published: June 2021

AMPA, NMDA, and kainate receptors belong to the ionotropic glutamate (iGluR) family of receptors located in the post‐synaptic neural membrane. Upon binding to glutamate, a major fast excitatory neurotransmitter, these channels activate and play an important role in synaptic plasticity, learning and memory. Mis‐regulation of these receptors has been linked to neurodegenerative diseases such as Alzheimer's and Parkinson's.

Our research is aimed at understanding the regulation of AMPA and NMDA receptors by the endogenous neurosteroids pregnenolone sulfate (PS) and 3α‐hydroxy‐5β‐pregnan‐20‐one sulfate (Pregas) for potential disease state applications. PS potentiates NMDA receptors containing NR2A or NR2B subunits, while inhibiting NMDA receptors containing NR2C or NR2D subunits and all other iGluRs. Likewise, Pregas negatively regulates all iGluRs. Our findings show that both neurosteroids bind to the amino terminal domain (ATD) and S1S2 domain of the NMDA NR2B and AMPA GluR2 subunits, but only bind to the ATD of the NDMA NR2D subunit. Intrinsic fluorescence studies confirm the binding of PS and Pregas to the subunit domains. Data from additional studies, including KD and Stern‐Volmer analysis, will be presented to further elucidate and differentiate binding.
A fluorescent glutathione analog for monitoring interactions of GST fusion proteins
Authors: Huff Hannah C , Gaines Carson R , Caldwell Benjamin D
Published: June 2021

Glutathione S‐Transferase (GST) has commonly been used as a carrier protein in the expression of fusion proteins due to its ease of purification. We are attempting to develop a generic system by which a single probe might be used to monitor many different GST fusion proteins rather than relying on specifically targeted probes which can add expense and time to studies. A probe targeted at GST may reduce the cost and time of screening multiple GST fusion proteins. A fluorescently labeled analog of glutathione was synthesized for monitoring the interaction of GST fusion proteins with target proteins. We will describe the synthesis of fluorescein labeled glutathione (GSH‐Fl) and the purification and characterization of the labeled GST substrate. Initial studies examining the binding of the GSH‐Fl to GST and GST fusions proteins using standard fluorescence and fluorescence anisotropy assays will be described, and its potential to measure protein interactions of GST fusion proteins with target proteins will be discussed.
Dietary Bovine Lactoferrin Stimulates Intestinal Proliferation in Piglets
Authors: Reznikov Elizabeth Alexandra , Comstock Sarah S , Contractor Nikhat , Donovan Sharon M
Published: June 2021

Human milk contains factors, such as lactoferrin(LF), that stimulate gastrointestinal growth and maturation. Herein, proliferative properties of bovine LF(bLF) were explored through 5‐ethynyl‐2′‐deoxyuridine(EdU) incorporation in vivo. Colostrum‐deprived pigs were fed formula containing 0.4(CON), 1(LF1) or 3.6(LF3) g bLF/L. On d7 or 14, pigs were intraperitoneally injected with 10mM EdU(1ml/kg body weight) 2h prior to euthanasia. Proliferation was assessed ex vivo in formalin‐fixed jejunal sections using Click‐iT EdU Alexa Fluor 555 Kit. Sections were imaged with a confocal microscope. Staining was quantified with AxioVision, where area positive for EdU(proliferation) was normalized by the area positive for DAPI(total nuclei). Formalin‐fixed jejunum sections were stained with haematoxylin and eosin for villus and crypt measurements. bLF did not affect body weight gain or intestinal weight or length. LF1 and LF3 pigs had a 1.5‐ fold increase in crypt cell proliferation compared to CON pigs. In LF3 pigs, crypt depth and area were greater(1.5‐fold on d7, 1.3‐ fold on d14) than in CON pigs. This study supports in vitro studies that showed LF could stimulate proliferation and is the first to show that physiologically‐relevant concentrations of milk‐borne bLF stimulate crypt cell proliferation in pigs suggesting LF plays a role in healthy GI development.(Funded by Pfizer Nutrition)

Grant Funding Source : Pfizer Nutrition
Hi‐5 Way: A multidisciplinary approach to secondary prevention of obesity in children 5 and under from underserved areas
Authors: Gorman Kathy A. , Lim-Miller Aimee , Oliver Lauren , Lenders Carine
Published: June 2021

There is evidence that most excess weight gained at puberty occurs before age 5. Rapid weight gain in infancy and early childhood is associated with obesity related complications later in life. Prevention of excess weight gain should begin in infancy and early childhood.

The Nutrition and Fitness for Life (NFL) program is a multidisciplinary pediatric weight management clinic that targets underserved populations. In 2009, NFL began a secondary prevention intervention for children ages 5 and under who are obese or rapidly gaining weight: Hi5‐Way. Our team includes a medical doctor, registered dietitian, and mental health provider. After an in‐depth assessment, we help children achieve healthier weight gain through a variety of family interventions.

Topics addressed include: role modeling, “table talk”, sleep/bedtime routines, meal/snack patterns, increasing active play, infant feeding, menu ideas, and picky eating. Material is modeled after Ellyn Satter's feeding principles and the WIC “Touching Hearts Touching Minds” program. Patients are referred to the BMC food pantry and community programs to promote health via increased access to resources. Frequency of clinic visits is tailored based on individual needs.

Conclusion

Secondary prevention of chronic diseases related to nutrition among underserved populations with young children is feasible. Support from the New Balance Foundation.
Breakfast and Ready‐to‐Eat Cereal Consumption is Associated with Improved Markers of Cardio‐metabolic Health in Adults: Results from National Health and Nutrition Examination Survey 2001 – 2008
Authors: Sanders Lisa M , Miller Kevin B , Fulgoni Victor
Published: June 2021

The objective of this study was to examine the association of breakfast, in particular ready to eat cereal (RTEC) consumption, with cardio‐metabolic health measures in adults 19–50 years (n=9292) using NHANES, 2001–2008. Using the first dietary recall, breakfast consumption was self‐defined by participants (n=6848) and any amount of RTEC for breakfast defined RTEC consumption (n=1148). Least square means ± SE were determined by regression analysis and odds ratios for risk factors were determined by logistic regression with adjustment for covariates. Compared to other breakfasts, RTEC breakfast was associated with a lower incidence (%±SE) of elevated BP (21±0.2 v 25±0.1, p<0.05), elevated blood glucose (27±0.2 v 34±0.1, p<0.003) and metabolic syndrome (20±0.1 v 25±0.1, p<0.007). RTEC breakfast was associated with lower LDL‐cholesterol (110.8±1.8 v 116.8±1.4, p<0.02) and lower fasting glucose (96.8±0.6 v 98.7±0.7, p<0.03) compared to breakfast skippers. RTEC breakfast consumers were also less likely to have elevated BP (OR=0.64; CI=0.50–0.82), elevated triglycerides (OR=0.69; CI=0.52–0.93), elevated fasting glucose (OR=0.57; CI=0.43–0.77), or metabolic syndrome (OR=0.50; CI=0.40–0.64). Consumers of other breakfasts showed a similar risk reduction, but odds ratios were slightly higher (0.70–0.85) than for RTEC (0.40–0.59). These data suggest breakfast consumption, particularly RTEC, is associated with improved markers of cardio‐metabolic health in adults. (Supported by Kellogg Company)
Korean high school male students’ recognition and use of nutrients‐enriched foods
Authors: Chang Kyung Ja , Park Jae-Ran
Published: June 2021

In order to investigate Korean high school students’ recognition and use of nutrients‐enriched foods, this cross‐sectional survey was conducted using a questionnaire. The subjects of this study were 397 male students in three different high schools in Incheon metropolitan city, Korea. Data were analyzed with SPSS 14.0 program. As for use of nutrients‐enriched foods, 69.0% of the subjects ate various nutrients‐enriched foods. The nutrients‐enriched food most eaten by the subjects was confectionery, ramen, milk/soybean milk, yoghurt/cheese and beverages in order. Only 7.6% of the subject use nutrients‐enriched foods after confirming that they were enriched with nutrients. The subjects acquired the most information on nutrients‐enriched foods from newspapers, magazines, TV and internet. As for reason for using nutrients‐enriched foods, most of the subjects answered that they were good for health. As for reason for using none of nutrients‐enriched foods, most of the subjects answered that they were not effective. Calcium and vitamin C were most favorite enriched nutrients and most of the subjects intended to use nutrients‐enriched foods. Therefore, nutrition education for use of nutrients‐enriched foods should be provided at school and specific regulations and law of nutrients‐enriched foods should be prepared to ensure the effective intake of nutrients‐enriched foods in Korea.
Child appetitive traits influence dietary intake in treatment seeking overweight children
Authors: Cahill Jodi M , Iacovino Juliette M , Kolko Rachel P , Bitow Naomi , Stein Richard I , Welch Robinson , Perri Michael G , Schechtman Kenneth B , Epstein Leonard H , Saelens Brian E , Wilfley Denise E
Published: June 2021

Appetitive traits, the individual response to internal and external eating cues, are related to weight status in children, but their influence on dietary intake has not been examined. The aim of this study was to determine the association between appetitive traits and diet in a treatment‐seeking sample of overweight children (ages 7–11; N=185). The Child Eating Behavior Questionnaire assessed appetitive traits. Dietary intake was determined using three 24‐hr recalls. Food approach scores were positively associated with sweetened beverage, total vegetable, and fried potato intakes (all p<0.05). In contrast, food avoidance scores were negatively associated with energy, carbohydrate, sugar, total vegetable, and non‐starchy vegetable intakes (all p<0.05). Associations differed between boys (n=70) and girls (n=115). In boys, the food approach trait enjoyment of food was associated with lower energy intake, while the food avoidance traits food fussiness and slowness in eating were related to higher energy intake (p<0.05). In girls, the food avoidance traits emotional under‐eating and slowness in eating were related to lower energy intake (p<0.05). These results indicate that appetitive traits do influence diet in overweight children and differ by gender.
Canned fruits, vegetables, beans and fish provide nutrients at a lower cost compared to fresh, frozen or dried
Authors: Kapica Cathy , Weiss Wendy
Published: June 2021

Discussions around increased fruit and vegetable consumption often emphasize ‘fresh,’ derived from the assumption that canned, frozen and dried may be lower in nutritional quality. The purpose of this study was to compare the nutrient content among canned, fresh, frozen and dried forms to the economic and time cost for serving edible portions of carrots, corn, green snap beans, mushrooms, peas, pumpkin, spinach, tomatoes, pears, peaches, pinto beans and tuna fish. Protein, fiber, potassium, vitamin C, vitamin A and folate content were from the USDA Nutrient Data Laboratory. The value of time spent on preparation was calculated based on the minimum wage. “Total cost per edible portion” was calculated by adding the average price per drained, prepared, edible serving to the cost of any waste and to the value on time spent during preparation. “Cost per nutrient” was calculated by dividing total cost per edible portion by the amount of each nutrient available in that portion. Canned fruits, vegetables, beans and tuna had the lowest total cost per edible portion across all categories. For most nutrients, the cost per nutrient for canned was less than or comparable to fresh, frozen or dried. Dietary recommendations should include canned foods as affordable, convenient sources of key nutrients. Funding provided by the Canned Food Alliance.
In utero bisphenol A exposure: Effects on metabolic homeostasis throughout the life‐course
Authors: Anderson Olivia S , Sanchez Brisa N , Peterson Karen E , Zhang Zhenzhen , Dolinoy Dana C
Published: June 2021

Animal studies have linked in utero and early postnatal bisphenol A (BPA) exposure to increased body weight but controversy exists about BPA's role as an obesogen given evidence for its effect on spontaneous behavior and cognitive function in absence of altered body weight. Utilizing isogenic a/a mice, we examined effects of in utero exposure of 50 ng (n=20), 50 ug (n=21), or 50 mg (n=18) of BPA/kg diet on offspring energy expenditure, activity, and body composition at 3, 6, and 9 mo of age using linear mixed regression adjusting for sex and light cycle. Results indicate that mg offspring had higher oxygen consumption at 3 mo (p=0.002) as well as ng female offspring at 9 mo (p=0.05) compared to control offspring (n=19). In females, ambulatory activity was increased in mg offspring at 9 mo (p=0.002), ug offspring at 3 and 9 mo (p=0.001 and 0.02), and ng offspring at 3 mo (p=0.02). Vertical activity was increased in mg offspring at 3 mo and in females at 9 mo (p=0.0006 and 0.02), and in ug offspring at 9 mo among females (p=0.003). Also, ng and ug offspring display marginally significant decreases in fat percentage and body weight. Our life course analysis indicates BPA is associated with hyperactive and lean phenotypes in a/a mice, underscoring the need to evaluate the effects of early BPA exposure in human clinical and population studies. This work is supported by NIH grants DK089503 and ES017524 and NIH/EPA grant P20 ES01817101/RD834800.
Assessing beans as a source of intrinsic protein and fiber on satiety in men and women with the Metabolic Syndrome
Authors: Randolph Jody M , Reverri Elizabeth J , Kappagoda C. Tissa , Park Eunyoung , Edirisinghe Indika , Burton-Freeman Britt
Published: June 2021

Multi‐pronged dietary strategies are gaining increasing attention as tools to manage appetite and provide metabolic and weight control in people with the metabolic syndrome (MS). The study aim was to assess the effect of beans and the fiber component of a diet on satiety and metabolic endpoints in people with the MS. In a 3‐arm randomized cross‐over design, 12 subjects with the MS ate a standard breakfast plus a side dish of either: whole beans (B), pasta matched for fiber (F, 13g), or pasta with no fiber added (NF, 0g). Meals were matched for energy and macronutrients. Subjective satiety (SS; by VAS), glucose, insulin and PYY3–36 were assessed over 5 h after meal consumption; subsequent intake was assessed from food records until midnight the next day. Main effects of meal and time were apparent for hunger (p=0.02), desire to eat (p=0.008), prospective consumption (p=0.0002), meal satisfaction (p=0.003), level of comfort (p=0.02), and level of contentment (p= 0.01). Meal B produced a greater level of comfort and contentment than F or NF, whereas F induced greater reductions in hunger, desire to eat, and prospective consumption compared to B or NF. Insulin was significantly reduced after meal B compared to F (p=0.03) or NF (p<0.0001), with no differences in glucose among meals. PYY3–36 was significantly greater after the B meal compared to F (p<0.0001) or NF (p=0.01). Beans appear to regulate appetite in patients with the MS.
The effects of BMI and ratings of hunger on perceived taste of food
Authors: Stickel Ariana Marie , Green Erin , Murphy Claire
Published: June 2021

The current study investigated psychophysical responses to foods as rated by participants with moderate Body Mass Index (BMI < 25, n= 15) as compared to participants with a high BMI (BMI ≥ 25, n= 25). The objective of the study was to identify disparities between high and moderate BMI groups, so future studies may investigate the mechanisms contributing to these differences. Pleasantness, intensity, liking, and wanting were rated on general Labeled Magnitude Scales (gLMS). Measures of hunger were self‐reported magnitude estimates and scores on the Three Factor Eating Questionnaire (factor three). Significantly higher flavor ratings of pleasantness and wanting by the moderate BMI group compared to the high BMI group were identified through independent samples t‐tests; e.g., pleasantness of chocolate was rated significantly higher by moderate than by high BMI participants. Correlations revealed greater association between hunger ratings and psychophysical responses in moderate than high‐BMI participants. Findings are consonant with recent fMRI data from our laboratory that suggest people with high BMI show less brain activation to sweet taste in reward areas than those with lower BMI. The results are compatible with a reward deficiency hypothesis. Supported by NIH grant #AG04085‐25 to Claire Murphy and NIH grant #MH065183 to Thereasa Cronan.
Effects of dietary carotenoids on steroid hormone status in male mice lacking carotene‐15,15′‐monooxygenase (CMO‐I) or carotene‐9′,10′‐monooxygenase (CMO‐II)
Authors: Smith Joshua W. , Ford Nikki A. , Clinton Steven K. , Erdman John W.
Published: June 2021

Higher serum levels of lycopene are inversely associated with prostate cancer incidence, while alterations in the gonadal hormones testosterone (T) and estradiol (E) modulate risk. Nine‐ to twelve‐week‐old male WT, CMO‐I KO and CMO‐II KO mice were fed either a control, lycopene beadlet‐ (LYC) or tomato powder‐supplemented (TP) diet for four days. Previously we reported that diet and genotype significantly interact, resulting in reduced testicular and total serum T in CMO‐I KO mice fed either TP or LYC, compared to WT mice. We hypothesized that CMO‐I KO genotype, but not CMO‐II KO genotype, would interact with LYC and/or TP to result in decreased serum E and alter testicular mRNA expression of 17β‐hydroxysteroid dehydrogenases (17β‐HSD) 2 & 3. Compared to WT, CMO‐I KO genotype significantly reduced the testicular mRNA expression of both 17β‐HSDs 2 and 3 (p<0.05). Serum E was unchanged, although there were trends towards significant genotype‐diet interactions in the CMO‐I KO/TP (increased; p=0.06) and CMO‐I KO/LYC (decreased; p=0.08) groups, respectively, compared to WT. No changes in serum E were seen in any CMO‐II KO mice. Further work is underway to expand upon these findings.

Grant Funding Source : NIH grant PHS‐1‐RO1 CA125384.
Biofortification of maize for nutritional security
Authors: Chaudhary Dharam Paul , Singh Sapna , Mandhania Shiwani , Srivastava Pallavi , Dass Sain , Kumar Ramanunjam S
Published: June 2021

Objectives

Maize is the staple food for a large part of population in this planet. The nutritional quality of maize was significantly improved by the development of quality protein maize which could be further enriched through biofortification of maize with carotenoids and β‐carotene. Keeping this in mind we have planned to characterize maize germplasm for protein quality, carbohydrate profile, total fat and carotenoids composition.

Methods

Forty elite maize inbreds were used for this study. The samples were dried, ground to powder and analyzed for protein quality, oil, starch, sugars and carotenoids composition and the data was further processed to calculate Euclidean coefficient of dissimilarity among genotypes.

Results

The results indicate that a significant number of inbreds posses quality protein. A few genotypes exhibit higher concentrations of starch. However, the starch is negatively correlated with oil content. A significant variation was observed in the total carotenoids composition. The inbreds could be grouped into five major categories based upon the Euclidean coefficient of dissimilarity in protein quality and carotenoids composition.

Conclusion

A significant variation was observed in the protein quality and carotenoids composition which could effectively be utilized in future breeding programme for the development of nutritionally improved maize hybrids.

Source of research support: The Indian Council of Agricultural Research, India.
The role of N‐linked glycosylation in the trafficking and transferrin binding of TfR2
Authors: Zhao Ningning , Enns Caroline A.
Published: June 2021

Transferrin receptor 2 (TfR2) belongs to the transferrin receptor‐like family and mediates cellular uptake of transferrin‐bound iron. TfR2 plays a key role in regulating iron homeostasis in the body. Mutations in this gene lead to a rare form of hereditary hemochromatosis. TfR2 is a type II protein with a single‐pass transmembrane region and four potential glycosylation sites on its ectodomain. The importance of glycosylation in TfR2 trafficking and function has not yet been studied. In the present study, we investigated the function of N‐linked glycosylation in the plasma membrane localization and transferrin binding of TfR2. We first used site‐directed mutagenesis to identify the glycosylation sites of TfR2. We found TfR2 is glycosylated at asparagines 240, 339 and 754, asparagine 540 is not glycosylated. By cell‐surface protein biotinylation and biotin‐labeled transferrin, we observed that without N‐linked glycan TfR2 can still traffic to the plasma membrane and bind to its ligand transferrin. Based on our analyses, we conclude that N‐linked glycosylation is dispensable for the cell‐surface expression and transferrin binding of TfR2.

Grant Funding Source: NIH
Vitamin D status of immigrant mothers and infants in Metro Vancouver
Authors: Li Wangyang , Barr Susan I , Green Tim J , Whiting Susan J , Chapman Gwen E
Published: June 2021

Recent studies suggest Canadian children from immigrant families may be at higher risk for rickets. However, little is known about the vitamin D status of immigrant mothers and infants in Canada. We measured 25‐hydroxyvitamin D (25OHD) in Asian and Middle Eastern immigrant (n=40) and White (n=40) mothers and their infants aged 2–4 months living in Vancouver (49°N). Mothers completed diet and sun exposure questionnaires. Mean 25OHD concentrations of immigrant mothers were lower than those of White mothers (68 vs 78 nmol/L; p=0.02), but 25OHD concentrations of their babies did not differ (82 vs 79 nmol/L; p=0.67). Only 1 White mother, 1 infant with a White mother, and 1 infant with a Middle Eastern mother had a 25OHD concentration indicative of severe deficiency (<25 nmol/L). However, 7 (18%) immigrant mothers and 2 (5%) White mothers (p=0.08) and 3 (8%) immigrants’ infants and 6 (15%) White infants (p=0.29) were vitamin D insufficient (<50 nmol/L). Multivariate analyses showed that ethnicity, body mass index, and vitamin D supplement use were significant predictors of 25OHD in mothers. Only supplement use was a significant predictor for infants. In conclusion, immigrant mothers were at marginally higher risk of vitamin D insufficiency than White mothers, but their infants had similar vitamin D status. (Supported by the Canadian Vitamin Class Action settlement)

Grant Funding Source: Canadian Vitamin Class Action settlement
Relationship between isoflavone intake and metabolic syndrome diagnostic components in Korean postmenopausal women
Authors: Bae Yun Jung , Kim Mi Hyun
Published: June 2021

Recent studies have suggested that natural agents such as isoflavones, resveratrol, and anthocyanin have beneficial effects on metabolic syndrome (MetS)‐related disorders. The objective of this study was to evaluate among isoflavone intake, urinary isoflavone level, and the risk of MetS among Korean postmenopausal women. The subjects included 46 MetS and control (n=60). The MetS risk score was determined by adding the number of risk factors such as waist circumference and high levels of triglyceride (TG), HDL‐cholesterol, glucose, and blood pressure (BP). Dietary isoflavone intake was not significantly different between MetS and control groups; however, the urinary daidzein level was significantly higher in MetS subjects compared to that of the controls. Subjects with high TG had higher urinary daidzein and isoflavone levels than those without such abnormalities. The correlation analysis indicated that dietary isoflavone intake had a positive relation with energy intake. After adjusted energy intake, there was no significant relationship among isoflavone intake, metabolic syndrome, and its components in Korean postmenopausal women. These results indicate that women who have more dietary isoflavones tend to have more energy. Furthermore, this study shows no significant direct relationship between isoflavone intake and risk of MetS in Korean postmenopausal women.
Monocarboxylate transporter‐1 genotype and breastfeeding status protect against elevated BMI in preschool aged Caucasian children
Authors: Wang Anthony An , Donovan Sharon M , Teran-Garcia Margarita
Published: June 2021

The gut microbiota has been implicated in the development of obesity in part through fermenting dietary fibers to short chain fatty acids (SCFAs). The monocarboxylate transporter‐1 (MCT‐1), coded for by the SLC16A1 gene, facilitates the uptake of SCFAs in the colon. The association between genetic variants of the SLC16A1 gene at rs4839273 and obesity phenotypes in Caucasian 2–5 year‐old children (n=128) in the STRONG Kids Program was studied. The marker was in Hardy‐Weinberg Equilibrium (χ2=0.38, p=0.54) and polymorphic information content was 0.34. Genomic DNA was obtained from saliva samples and was genotyped by fluorescence polarization. After adjusting for age, sex, and breastfeeding (BF) status, children with the CC genotype had lower BMI than children with CA or AA alleles (15.3 ± 0.6 vs. 16.6 ± 0.1, p=0.02). Breastfeeding status moderated this effect; individuals with CC genotype that were BF had lower BMI than those with CA or AA alleles who were not BF (15.9 ± 0.4 vs. 16.7 ± 0.2, p=0.05). Thus, genetic variation within the SLC16A1 gene was associated with BMI of the children in our study and this relationship was mediated by BF status.

Grant Funding Source : UI Research Board
Regulation of the PBMCs gene expression profile with the Western dietary pattern in healthy men and women
Authors: Bouchard-Mercier Annie , Paradis Ann-Marie , Rudkowska Iwona , Lemieux Simone , Couture Patrick , Vohl Marie-Claude
Published: June 2021

Diet regulates gene expression. The objective is to examine gene expression in relation with the Western dietary pattern.

Methods

30 Caucasians (13 males; 17 females) were recruited. Dietary patterns were derived from a food frequency questionnaire using factor analysis. RNA was extracted from peripheral blood mononuclear cells (PBMCs), when subjects were in the fasting state. Expression levels of 47 231 mRNA transcripts (> 31000 genes) were assessed using the Illumina Human‐6 v3 Expression BeadChips®. Microarray data was processed with Flexarray software and pathway analysis was assessed using Ingenuity Pathway analysis (IPA).

Results

The Western pattern was characterised by high intakes of refined grain products and meats. In subjects with high versus low scores for the Western pattern, 193 transcripts were upregulated and 197 were downregulated. IPA reveals that networks related to inflammatory response, immunological diseases and cancer were different in subjects with high versus low scores for the Western pattern.

Conclusion

Studying gene expression profiles according to dietary patterns may help to understand the overall effect of nutrition in chronic diseases development. CIHR (MOP229488)
Studies on the Biochemical Effect of a Local Energy Drink (KPE) on Rats
Authors: ogunlabi olugbenga owolabi , Banjoko Immaculata Oluyemisi , Adebawo Olugbenga O , Adenuga Gbenga A.
Published: June 2021

KPE was produced from a blend of kola nut extract and fresh pineapple juice. 24 matured male Wister strain albino rats (190g – 230g) were used for the experiment. For the study, rats were divided into four (4) groups and treated for 4 weeks after 2 weeks acclimatisation as follows: experimental group 1 received 5–7 ml KPE/day, experimental group 2 received 10–14 ml KPE/day, and standard group 3 received 5–7 ml ‘power horse’(a commercial energy drink)/day while control group 4 was given ordinary water. The fasting blood glucose, body weight, feed intake and water intake were measured. The rats were sacrificed and blood samples were taken and used for liver and kidney functions analysis. The liver, kidney, heart and testis were excised and used for tissue histopathological examination. The result of the study indicated that fasting blood glucose was significantly elevated in groups 1, 2 and 3 when compared with group 4 (control). Body weight was significantly increases in the control (group 4) compared with groups 1, 2 and 3 Level of serum transaminases (alanine aminotransferase and aspartate aminotransferase) was significantly higher in groups 1, 2 and 3 than in group 4. Serum urea level in group 3 and group 1 was not significantly greater than the control but group 2 showed significantly high serum urea level when compared with the control. Na+ concentration in all the groups was not significantly different while level of K+ was significantly higher in groups 1, 2 and 3 when compared with the control. The testis and liver of animals in groups 1, 2 and 3 were significantly enlarged but histopathological examination of tissues revealed no significant cellular abnormality. Result of the study suggests that chronic administration of KPE may alter liver and kidney function.
ACTION OF HYPERTONIC SALINE SOLUTION (NaCl 7,5%) IN PULMONARY FIBROSIS IN A RODENT MODEL OF ENDOTOXEMIA
Authors: Petroni Ricardo Costa , Biselli Paolo , Martins Milton , Csaba Szabo , Soriano Francisco
Published: June 2021

Hypertonic saline solution (HSS, NaCl 7,5%) has shown to modulates immune function favorably and decrease pulmonary injury triggered by endotoxic shock. Therefore, our objective was to investigate the effects of HSS on the mechanism involved in pulmonary fibrosis, in an experimental model of endotoxemic shock. Wistar rats (220–260 g) received lipopolysaccharide ‐ LPS (10mg/kg, intraperitoneal,ip) and volume after 15 minutes. The animals were assigned in four groups (n=7 per group): control group (not subjected to LPS); LPS group (injected with LPS 10mg/kg i.p); HSS group (injected with LPS and treated with hypertonic saline, 4 mL/Kg) and NS group (injected with LPS and treated with normal saline, 34 mL/kg). At 24h after treatment, pulmonary mechanics, type I collagen expression, metalloproteinase 9 expression and activity, and focal adhesion kinase (FAK) were measured. Normal saline increased pulmonary resistance and elastance, compared to other groups (p<0.05). HSS inhibited collagen expression compared to LPS and NS groups and prevented pulmonary injury by decreased MMP9 activity in tissue (p<0.05). Expression of FAK was decreased in HSS groups compared to LPS and NS groups (P<0.05). Therefore, we concluded that treatment of endotoxemic shock with HSS solution can decreased pulmonary fibrosis and injury through FAK pathway. FAPESP 2009/07478‐8
In vivo study: Developmental alterations in primary rat neuronal and glial cell cultures following prenatal cocaine exposure
Authors: Johnson Marquita L , Womble Tracey , Jackson Tanise , Goodman Carl B
Published: June 2021

Evidence has shown that prenatal cocaine exposure to the developing fetus is associated with neurological and behavioral abnormalities. These alterations are attributed to structural damage in the brain as well as neurodevelopment with no significant underlying mechanisms. Since the effects of prenatal cocaine exposure on glial and neuronal development are largely unclear, there is a need to establish an appropriate animal model to evaluate cocaine's antecedent role in the occurrence of neuronal and glial cell maturation. The present study addresses the hypothesis that chronic prenatal cocaine exposure alters the maturation and function of neurons and glial cells in the offspring. Timed pregnant Sprague‐Dawley rats were treated with 40 mg/kg of cocaine on gestational day 10–21. The brain tissue of the pups was harvested to yield pure neuronal and glial cell cultures. ICC analysis confirmed the successful isolation of primary neurons and glial cells, as well as morphological differences between the control and treatment pups among these cell types. Western Blot analysis determined a variation of protein expression. A cytokine array was used to identify cytokine expression levels. These studies suggest that cocaine alters growth, maturation, and function of growing neuronal and glial cell cultures. Excessive glial cell activation can cause harm to neurons in regards to their development and plasticity.
Dry Ethanol Gas Testing of Personal Use Breathalyzers
Authors: Blackburn Mollie , Lee J. Rex
Published: June 2021

There are approximately 13,000 fatalities resulting from alcohol‐impaired accidents each year. The use of accurate personal breath alcohol analyzers (BAA) could help reduce this number. The purpose of this study was to determine the accuracy of 5 consumer models of semi‐conductor type breath alcohol analyzers. For this study, each instrument was tested 10 times at three concentrations (0.035%, 0.070% and 0.087%) of a dry gas ethanol standard (National Institute of Standards‐traceable) at both room and human breath temperature. An evidentiary BAA (Lifeloc FC20) was also used as an internal control for the reference standard. The results of the study showed that none of the 5 instruments were accurate at any of the three concentrations tested. The accuracy was not affected by the change in temperature. Therefore, we concluded that none of the 5 breath alcohol analyzers studied could accurately inform the user of their breath alcohol status. Since these products are advertised as accurate, the advertisement appears to be misleading. Given the lack of accuracy found in this study, these breath alcohol analyzers would most likely not reduce the fatalities from alcohol‐impaired accidents.
Eating high fat chow enhances sensitivity, but does not impact the development of sensitization, to apomorphine‐induced yawning in rats
Authors: Baladi Michelle G , Thomas Yvonne M , France Charles P
Published: June 2021

Eating high fat chow can impact the effects of direct‐acting dopamine receptor agonists; however, it is unclear if eating high fat chow alters sensitization that sometimes develops from repeated administration of dopamine receptor agonists. This study examined whether eating high fat chow affects the potency of apomorphine to induce yawning or alters the development of sensitization to apomorphine. Quinpirole‐induced yawning was evaluated to see whether sensitization to apomorphine confers cross‐sensitization to quinpirole. Male Sprague‐Dawley rats (n=6/group) had free access to high fat (34.3% fat) or standard chow (5.7%). Apomorphine and quinpirole yawning dose‐response curves were shifted leftward in rats eating high fat chow compared with rats eating standard chow. Sensitivity to apomorphine increased over once‐weekly testing (sensitization) and that increase was not different between rats eating standard or high fat chow; conditioning to the testing environment appeared to enhance yawning. Rats eating standard chow and sensitized to apomorphine were cross‐sensitized to quinpirole‐induced yawning; cross‐sensitization was not evident in rats eating high fat chow. These results demonstrate that sensitization can develop to dopamine agonist‐induced yawning and that both drug history and feeding condition can impact the potency of drugs acting on dopamine systems. CPF supported by K0DA17918.
Pharmacological differences between two mechanistically similar drug effects, nitrous oxide (N2O)‐ and hyperbaric oxygen (HBO2)‐induced antinociception in mice
Authors: Dupic Christine M. , Zhang Yangmiao , Shirachi Donald Y. , Quock Raymond M.
Published: June 2021

N2O‐ and HBO2‐induced antinociceptive effects both appear to involve an NO‐cyclic GMP‐protein kinase G‐dependent release of dynorphin that activates kappa opioid receptors. This study was conducted to further differentiate between their mechanisms of action. Antinociceptive responsiveness of male NIH Swiss mice to N2O and HBO2 was assessed using the acetic acid‐induced abdominal constriction test. Pretreatment drugs were the CB1 antagonist AM 251, the angiotensin‐converting enzyme (ACE)‐inhibitor captopril and the GABA antagonist SR‐25531. Control mice responded to N2O and HBO2 with a robust antinociceptive effect. N2O‐induced antinociception was antagonized by AM 251 but was resistant to antagonism by captopril and SR‐95531. HBO2‐induced antinociception was dose‐dependently antagonized by captopril but resistant to antagonism by AM 251 and SR‐95531. These results suggest that there are mechanisms that differentially modulate the antinociceptive responses to N2O and HBO2, notably involvement of ACE and CB1 but not GABA mechanisms. (This research was supported by NIH Grant GM‐77153, the Allen I. White Distinguished Professorship and an institutional Summer Undergraduate Research Fellowship (SURF) Award from the American Society for Pharmacology and Experimental Therapeutics).
Regulation of protease‐activated receptor‐4 signaling and trafficking
Authors: Smith Thomas Horace , Trejo JoAnn
Published: June 2021

Protease‐activated receptors (PARs) are G‐protein coupled receptors (GPCRs) that elicit cellular responses to thrombin. PARs play pivotal roles in hemostasis, thrombosis, and inflammatory responses to tissue injury. PAR activation is unique compared to other GPCRs. Thrombin cleaves the receptor's N‐terminus to reveal a “tethered ligand” that binds in cis to the receptor, resulting in G‐protein signaling. Due to the irreversible nature of PAR activation, tight regulatory mechanisms are required to govern their signaling. Thrombin signals predominantly through the activation of PAR1 and PAR4. Most cell types express more than one PAR. Thus, delineating the mechanisms of PAR4 signal regulation is essential to understanding the regulation of thrombin signaling. The goal of this study is to define the mechanisms of PAR4 internalization. To identify the mechanism of PAR4 internalization we depleted cells of certain adaptor proteins by siRNA, which revealed that the clathrin adaptor protein complex‐2 (AP‐2) plays a necessary role in this process. Immunofluorescence microscopy experiments revealed that activated PAR4 colocalizes with clathrin and is trafficked through early endosomes, and late endosomes/lysosomes. This study highlighted several other key aspects of PAR4's signal regulation that will be presented. Supported by NIH R01 HL073328.
Activated protein C promotes endothelial barrier protection through biased PAR1 signaling mediated by β‐arrestin and dishevelled‐2 scaffolds
Authors: Soh Unice JK , Trejo JoAnn
Published: June 2021

Thrombin, a coagulant protease contributes to inflammatory responses associated with vascular injury and thrombosis. Activated protein C (APC), an anti‐coagulant protease reduces thrombin generation and promotes anti‐inflammatory signaling in endothelial cells. Thrombin and APC signal through the G‐protein coupled protease‐activated receptor1 (PAR1) but elicit opposite effects on endothelial barrier function. The mechanism of APC‐activated PAR1 signaling in endothelial barrier protection is poorly understood. In human endothelial cells, APC activates caveolar PAR1 populations to promote barrier protective responses independent of Gαi signaling. PAR1 is precoupled to β‐arrestins and co‐segregate in caveolin‐1 enriched fractions. APC‐activated PAR1 promotes β‐arrestin‐dependent recruitment and activation of dishevelled‐2 (Dvl‐2), a key mediator of the Wnt/Frizzled signaling pathways. Depletion of β‐arrestins or Dvl‐2 resulted in the loss of APC‐induced Rac1 activation and protection against thrombin‐activated endothelial barrier permeability but not thrombin‐induced RhoA signaling. These findings indicate that PAR1 utilizes β‐arrestins to promote biased signaling and unveils a distinct pathway where β‐arrestins and Dvl‐2 scaffolds promote APC effects on endothelial barrier protection. Funded by R01 HL073328, UC TRDRP 20FT‐0074.