bluuebunny/crossref_metadata_2025 huggingface dataset:
Determining the role of melanopsin C‐tail in deactivation and trafficking using chimeric constructs (596.7)
Authors: Ortiz Elelbin , Cameron Evan , Robinson Phyllis
Published: June 2021

Melanopsin is a non‐image forming G protein‐coupled receptor expressed in intrisically photosensitive retinal ganglion cells in the vertebrate retina. These cells are involved in functions such as the photoentrainment of circadian rhythm and the pupillary light reflex. The deactivation of melanopsin occurs through the phosphorylation of the C‐tail followed by the binding of a β‐arrestin molecule. The binding of β‐arrestin allows for endocytosis of GPCRs after inactivation. However, preliminary data suggests that melanopsin is not internalized. We hypothesize that the C‐tail of melanopsin plays a key role in the prevention of internalization.

Angiotensin II type 1A receptor (ATII1AR) and β‐2‐andrenergic receptor (B2AR) are GPCRs that bind β‐arrestin but differ in their trafficking via endocytic vesicles. In this study, the melanopsin C‐tail was replaced with either ATII1AR and B2AR C‐tail through cloning. The construction of the chimeras has been confirmed by DNA sequencing.

Our next step is to introduce our plasmids into Human Embryonic Kidney (HEK) cells. These constructs will then be assayed for trafficking using immuno flow cytometry and immunohistochemistry. We hypothesize that the chimeric constructs will undergo endocytotic pathways similar to ATII1AR and B2AR. This would allow us to conclude that the melanopsin c‐tail is essential in maintaining melanopsin in the cell membrane.

Grant Funding Source: Supported by HHMI, MARC U*STAR Program at UMBC, and a National Eye Institute grant to P.R.R.
Clathrin and its adaptor AP1 are required for apical sorting of GPI‐anchored proteins in biosynthetic and recycling routes in MDCK cells (598.4)
Authors: Watanabe Reika , Castillon Guillaume
Published: June 2021

To fulfill their functions, epithelial cells establish asymmetric protein distributions in apical and basolateral plasma membranes. To date, the role of clathrin and its adaptor, the AP1 complex, has been documented exclusively for basolateral protein trafficking in mammalian epithelial cells. We found that the apical localization of endogenous glycosylphosphatidylinositol (GPI)‐anchored proteins is severely affected by gene silencing of the clathrin heavy chain and components of the AP1 adaptor complex in MDCK cells. In these cells, apical targeting is affected not only for GPI‐ anchored proteins but also for endogenous secretory proteins and the apical‐destined t‐SNARE, syntaxin 3. Analysis of apical secretion of lysoGPI‐APs reveals that the biosynthetic delivery of GPI‐APs toward apical plasma membrane is impaired in these cells. Furthermore, apical recycling and transcytosis from basolateral to apical membranes of GPI‐APs are also impaired in these cells. Our results formally demonstrate the importance of biosynthetic and endocytic routes for establishment and maintenance of apical localization of GPI‐APs in polarized MDCK cells.

Grant Funding Source: Swiss National Sciences Foundation Assistant Professorship
Stearoyl‐CoA desaturase‐3 mediates the regulation of adipose and hepatic murine lipid metabolism (605.1)
Authors: Bond Laura , Burhans Maggie , Ntambi James
Published: June 2021

Stearoyl‐CoA desaturase (SCD) is a critical enzyme involved in de novo lipogenesis and catalyzes the Δ9 desaturation of saturated fatty acids. Four SCD isoforms have been identified in mouse (SCD1‐4); SCD isoform 3 exhibits unique substrate specificity and tissue expression. While SCD1, 2, and 4 convert palmitate and stearate to palmitoleate and oleate, respectively, SCD3 preferentially desaturates palmitate. SCD3 expression is restricted to skin, harderian glands, preputial glands and differentiating adipocytes. To determine the role of SCD3 in murine lipid metabolism, we created a mouse model in which SCD3 is globally knocked out. Despite the limited tissue expression of this isoform, SCD3‐/‐ mice exhibit a global phenotype when fed a lipogenic low‐fat high‐carbohydrate diet. In contrast to SCD1 deletion, which protects mice from diet‐induced obesity and hepatic steatosis, deletion of SCD3 deletion predisposes mice to increased hepatic triglyceride accumulation under high‐carbohydrate feeding compared to SCD3flox mice. Gene expression studies in white adipose tissue reveal that SCD3 deletion and high‐carbohydrate feeding cause a transcriptional remodeling of subcutaneous white adipose tissue. Expression of genes involved in de novo lipogenesis, fatty acid oxidation, and uncoupling are simultaneously increased in the subcutaneous depot of SCD3‐/‐ mice.

Grant Funding Source: Supported by NIH National Research Service Award T32 GM07215.
Catalytically important residues in the S. cerevisiae lysophospholipid acyltransferase, Lpt1p (605.15)
Authors: Oelkers Peter , Renauer Paul
Published: June 2021

Members of the membrane bound o‐acyltransferase family mediate reactions in phospholipid and triglyceride synthesis, cholesterol esterification, and protein modification. Aside from a conserved histidine, the amino acids critical for catalysis in this family remain to be conclusively determined. Thus, the reaction mechanism remains undetermined. We used bioinformatics to identify residues highly conserved among homologs spanning three kingdoms and to perform site directed mutagenesis of the respective residues in the S. cerevisiae homolog, LPT1. Eight residues were identified which have R‐groups containing hydroxyl, carboxyl, and primary amine groups. Initial assays suggest that an acid / base mechanism is utilized by Lpt1p to esterify lysophosphatidylcholine with oleoyl‐CoA. To further examine Lpt1’s mechanism, utilization of three 18‐carbon monounsaturated acyl‐CoA species with differing double bond locations was assayed in vitro. Interestingly, location of the double bond at position Δ11 lowered the apparent Vmax by six fold and at position Δ6 lowered it two fold compared to the Δ9 species. No activity was detected with a 24‐carbon, Δ9 acyl‐CoA species. Together, these data suggest in Lpt1p, the acyl‐CoA substrate lies in a protein furrow, uniquely suited to accommodate acyl‐CoA species with less than 24 carbons and a double bond nine carbons from the thioester.

Grant Funding Source: Supported by the University of Michigan Office of Research and Sponsored Programs
Identification of a putative perilipin 5 splice variant (606.6)
Authors: Hardy Cara , DuBreuil Daniel , Hubbell Theresa , Tansey John
Published: June 2021

Cells store neutral lipid in lipid storage droplets. These organelles have a coating of regulatory proteins including the perilipins, a family of related proteins which regulate and scaffold lipolytic enzymes. At least one of these proteins (perilipin 1) has been shown to have several truncated forms that arise through differential mRNA splicing. Using western blotting we have identified a truncated form of perilipin 5 we have termed perilipin 5B. This protein is found in murine tissue as well as cultured cells. Using RT‐PCR we have identified multiple amplicons suggesting that perilipin 5B contains a read through of intron 8 of the murine perilipin 5 gene. shRNA studies conducted in cultured cells result in decreased levels of the protein. In addition, immunofluorescence microscopy revealed that perilipin 5B is unable to bind to mature lipid droplets, but does form distinct puncta in the cytosol. Taken together, this data suggests that perilipin 5B is a splice variant of perilipin 5 found in oxidative tissues but is unable to bind to large lipid droplets. The role of this protein in neutral lipid metabolism remains unclear.
Discovery of novel monosaccharides in animal glycans: natural occurrence of N‐glycolylhexosamines (607.6)
Authors: Bergfeld Anne , Lawrence Roger , Diaz Sandra , Pearce Oliver , van Vleet Jeremy , Esko Jeffrey , Choudhury Biswa , Varki Ajit
Published: June 2021

Metabolism of the major mammalian sialic acid N‐glycolylneuraminic acid (Neu5Gc) generates N‐glycolylglucosamine (GlcNGc) as a byproduct of its degradative pathway (1). Mammalian cells cultured with exogenous chemically synthesized GlcNGc were found to synthesize UDP‐GlcNGc (2). Chemically synthesized N‐glycolylgalactosamine (GalNGc) was also utilized by mammalian cells to synthesize UDP‐GalNGc and UDP‐GlcNGc, and these served as precursors for assembly of most major glycan classes (3). We asked if these novel reactions could also happen naturally. Comparing wild‐type with Neu5Gc‐deficient mice, we demonstrate that Neu5Gc metabolism is indeed a source of naturally‐occurring UDP‐GalNGc and UDP‐GlcNGc. These endogenously derived N‐glycolylated aminosugars are also naturally incorporated into animal glycans such as chondroitin sulfates (CS), O‐glycans, and N‐glycans. In contrast, humans lack Neu5Gc biosynthesis and should therefore be devoid of GalNGc and GlcNGc in glycans, and also be deficient in any as yet unknown functions these glycans may mediate. Comparisons of human and chimpanzee samples indeed show the predicted difference. However, there is detectable trace amounts of GalNGc in human CS, likely derived from Neu5Gc‐containing animal‐derived foods, particularly of mammalian origin.

(1) Bergfeld et al., J Biol Chem. 2012 Aug 17;287(34):28865‐81 (2) Macauley et al., J Biol Chem. 2012 Aug 17;287(34):28882‐97 (3) Bergfeld et al., J Biol Chem. 2012 Aug 17;287(34):28898‐916

Grant Funding Source: This work was supported by NIH grant R01 GM32373.
Role of PKG‐I in differentiation of bone marrow‐derived mesenchymal stem cells (BM‐MSCs, OP9 cells) into lipid‐accumulating fat cells through RhoA phosphorylation, IRS‐1 and PI3‐kinase in insulin signaling pathway (609.18)
Authors: Gorjala Priyatham , Wong Janica , Constantino Benjamin , Johlfs Mary , Coffman Renee , Rosenberg Harry , Fiscus Ronald
Published: June 2021

3T3‐L1 cells have been used as a model for obesity and insulin response studies. A major limitation in using 3T3‐L1 has been a minimum two week time period required for allowing the cells to differentiate. Recently OP9 cells (bone marrow‐derived mesenchymal stem cells) have been found to differentiate into adipocytes within two days and express late markers of adipocyte differentiation. In an earlier study we found that PKG‐Iα is necessary for OP9 cell survival, proliferation and migration. Further, we have used BM‐MSCs as a model to study the mechanism of Insulin resistance and the role of PKG‐Iα in the pathway regulation. Recent studies show that serine‐phosphorylation of IRS (insulin‐receptor‐substrate) shortens its half‐life and decreases its association with PI3‐kinase. Our laboratory has shown that two isoforms of PKG‐I (PKG‐Iα and PKG‐Iβ) differ dramatically in their cellular functions. BM‐MSCs express only PKG‐Iα. Interestingly, type‐1 and type‐2 diabetes are associated with dramatic decreases in expression and kinase‐activity of PKG‐Iα. In the present study, we investigated the ability of BM‐MSCs to differentiate into lipid‐accumulating adipocytes in response to high‐insulin and the potential involvement of other insulin‐signaling‐proteins, PKG‐I, RhoA, IRS‐1 and PI3‐kinase (PI3K). We found that when these cells differentiate into lipid‐droplet‐containing cells the process is associated with dramatic decreases (upto 50%) of protein expression levels of PKG‐I, IRS‐1, PI3K and a decrease in RhoA serine‐188‐phosphorylation (site phosphorylated and regulated by PKG‐I). These changes in PKG‐I‐catalyzed phosphorylation of RhoA and decreased levels of IRS‐1 and PI3K may play a key role in the differentiation of BM‐MSC into fat cells, making them a useful model for adipogenesis, obesity and type‐2 diabetes.
CaM kinase activation and cancer cell growth are controlled by AKAP7 (613.4)
Authors: McFarland Hannah , Dodge‐Kafka Kimberly , Schmitt John
Published: June 2021

Estrogen (E2) activates the calcium/calmodulin‐dependent protein kinases (CaM Kinases) in breast cancer cells. The CaM Kinases regulate ERK leading to transcription and cell growth. CaM Kinase activation of ERK may be blocked by PKA phosphorylation and inhibition of CaM KK. The ability of PKA to phosphorylate its cellular targets may be dictated by protein kinase A anchoring proteins (AKAPs). Hormones that activate PKA, such as vitamin D (VitD) and epinephrine (Epi), may utilize AKAPs to facilitate the assembly of protein complexes and signal transduction. AKAP7 participates in intracellular calcium signaling and therefore may regulate CaM Kinases. Our goal was to evaluate the role of AKAP7 in regulating E2 activation of the CaM KK and CaM KI cell growth pathway in MCF‐10, MCF‐7 and MDA‐MB‐231 cells. Our results suggest that E2 treatment of MCF‐7 cells activates CaM KI, an effect that is blocked by the CaM KK‐inhibitor STO‐609 but not by the ERK‐inhibitor U0126. Interestingly, purified GST‐RII subunit of PKA pulled down both CaM KK and CaM KI from MCF‐7 cells. Similarly, precipitation with STag‐AKAPs showed that CaM Kinases bind to AKAP7 but not AKAP5, an effect that is enhanced with E2. Purified AKAP7 directly binds purified CaM KK, and endogenous AKAP7 is expressed in MCF‐10, MCF‐7, and MDA‐MB‐231 cells. Precipitation of endogenous AKAP7 revealed a robust association with CaM KK in E2‐, but not in STO‐609‐ or VitD‐treated cells. Similarly, VitD pre‐treatment blocked E2’s ability to increase CaM KI phosphorylation and activation. Interestingly, siRNA knockdown of AKAP7 increased basal ERK phosphorylation and prevented E2’s activation of ERK. Finally, E2‐stimulated cell growth was inhibited by pre‐treatment with VitD, STO‐609, and the AKAP inhibitor, Ht‐31. Taken together, these results suggest that AKAP7 directly associates with CaM KK in a complex with CaM KI, an effect that is blocked by VitD via PKA’s inhibition of CaM KK. Finally, VitD and Ht‐31 inhibit growth in breast cancer cells.

Grant Funding Source: Supported by M.J.Murdock Charitable Trust #2011267
Multiploidy formation in mitotic DNA damage response (616.4)
Authors: Hyun Sun‐Yi , Kim Hyo‐Joo , Jang Young‐Joo
Published: June 2021

The maintenance of genomic stability is significantly important in order to transmit the correct genome information to daughter cell. For defense from DNA damage that induces genomic instability, cells establish a checkpoint pathway which recognize DNA damage and arrest cell cycle. When DNA damage generated in mitotic HeLa cells, mitotic kinases are inactivated by ATM/Chk1/PP2A‐manner, and cells progressed into 4N‐DNA G1 phase. After then, cells are accumulated in 8N‐DNA, suggested that the formation of multiploidy during long‐term response on mitotic DNA damage was induced by cytokinesis failure and re‐replication. Cells with p53‐deletion or ‐inactivation is re‐replicated, whereas cells expressing p53‐wild type arrest before initiation of replication cycle by G1 checkpoint. p53 normal cells are removed by apoptosis at earlier stage of mitotic DNA damage in compare with cells with p53‐deficiency or inactivation of p53. These data suggest that although p53 does not interrupt pre‐RC assembly, p53 might be a key factor to block re‐replication by G1 checkpoint activation in mitotic DNA damaged cells. This research was supported by Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Science, ICT and Future Planning (NRF‐2013R1A1A3007213).
The impact of a multidisciplinary functional genomics project on the biochemistry and molecular biology curriculum at Otterbein University (618.29)
Authors: Tansey John , Bennett Jennifer , Hayes Carrigan
Published: June 2021

An interdisciplinary study examining the roles of c‐di‐GMP and its associated phosphodiesterases with respect to gene expression, cell signaling, and enzyme kinetics was incorporated into multiple courses in Otterbein University’s Biochemistry and Molecular Biology (BMB) program. Undergraduate science classes often appear to students to be detached from one another; this inquiry‐based project was intended to demonstrate the interplay between multiple disciplines and to provide a research experience within the Otterbein BMB curriculum. In Physical Chemistry 1, cyclic di‐GMP phosphodiesterase activities in crude lysates from wild‐type, heat‐inactivated wild‐type, and mutant cells were examined and quantified via kinetic assays. In Microbial Genetics, a three week intensive course offered during the January term, students completed a mini‐project using the same wild‐type and mutant cells. In this project, students examined potential targets of c‐di‐GMP signaling through semi‐quantitative PCR. Student learning was assessed primarily via pre‐ and post‐lab surveys; pertinent questions on pre‐lab assignments and subsequent exams were also evaluated. Preliminary data indicate that students demonstrated an increased understanding of content‐based objectives (for instance, enzyme specificity and kinetic experiment design in Physical Chemistry) following relevant labwork and class projects. Students across multiple classes reported that the project increased their understanding of the scientific process and their awareness of the multidisciplinarity of the research questions involved.
Small World Initiative: crowdsourcing research of new antibiotics to enhance undergraduate biology teaching (618.41)
Authors: Barral Ana Maria , Makhluf Huda , Soneral Paula , Gasper Brittany
Published: June 2021

The Small World Initiative (SWI) developed at the Center for Scientific Teaching at Yale uses an undergraduate research course approach to crowdsource the search for new antibiotics. The SWI framework has both a research and an educational component. Research consists of culturing soil microbes, screening for antibiotic activity against pathogens, 16s rRNA gene sequencing, and extracting metabolites. The educational component is embedded in the research framework as students take ownership of the project that includes experimental design, data analysis and presentations, while learning core biological concepts within a number of different introductory courses. Giving the students the opportunity to experience the excitement and process of research in the first years of college provides a positive, realistic reflection of science, resulting in higher retention rates within STEM fields. SWI is currently piloted at 25 diverse institutions from around the country, including community colleges and non‐traditional universities. In the pilot phase, student attitudes about learning science, experiences as a scientist, and relationship to a laboratory research project are evaluated using pre‐ and post surveys. We present the framework of the SWI and different implementation strategies as well as preliminary results of both isolation of antibiotic producers and student responses.

Grant Funding Source: Supported by The Leona M. and Harry B. Helmsley Charitable Trust, Davis Educational Foundation, HHMI
Assessing implementation fidelity of a community‐based infant and young child feeding intervention in Ethiopia identifies gaps in delivery that limit reach to communities (624.11)
Authors: Kim Sunny , Ali Disha , Kennedy Andrew , Tesfaye Roman , Rawat Rahul , Menon Purnima
Published: June 2021

Implementation fidelity is defined by adherence to intervention design, exposure or dose, quality of delivery, and participant responsiveness. As part of the process evaluation (PE) of Alive & Thrive, a large‐scale initiative to improve IYCF, our study assessed the four elements in training of frontline workers (FLWs), delivery of program tools and messages, and supervision and feedback. Data from a qualitative study among three levels of FLWs (n=54 in 2012), i.e. supervisors, health extension workers (HEWs), and community volunteers; and baseline and PE surveys with FLWs (n=225 in 2010, n=504 in 2013) and mothers (n=1,481 in 2010, n=750 in 2013) in two regions (Tigray and SNNPR) were analyzed to examine fidelity of key program components. There was strong adherence to intended cascading format (i.e. information transfer from higher to lower FLW levels) and high exposure to training (95% HEWs and 94% volunteers in Tigray, 68% and 81% respectively in SNNPR). Training quality, assessed by IYCF knowledge and self‐reported capacity, was high and increased since baseline. Job aids were used regularly by most supervisors and HEWs, but only 54% of volunteers in Tigray and 39% in SNNPR received them. Quality of program message delivery was lower among volunteers, and aided recall of key messages among mothers was also low. Although supervision exposure was high, frequency was irregular with weak content fidelity. There is evidence of strong fidelity in training and delivery of tools and messages at higher FLW levels, but gaps in the reach of these to volunteers and mothers and variability between regions limit the potential for intervention impact.

Grant Funding Source: Supported by: Bill & Melinda Gates Foundation, through Alive & Thrive, managed by FHI360
Factors associated with growth in disadvantaged preschoolers attending daycares in Salvador, NE Brazil (624.26)
Authors: Lander Rebecca , Williams Sheila , Costa‐Ribeiro Hugo , Mattos Angela , Barreto Danile , Houghton Lisa , Bailey Karl , Lander Alastair , Gibson Rosalind
Published: June 2021

Growth of preschool children from poor urban households of NE Brazil can be affected by multiple factors through complex pathways. Structural equation modelling examined the direct and indirect inter‐relationships between socio‐demographic, biological, environmental, and nutritional factors on z‐scores for height‐for‐age (HAZ) and body‐mass‐index (BMIZ) in a cross‐sectional study of 264 disadvantaged preschoolers aged 3‐6 years in 7 philanthropic daycares in Salvador. Statistically significant (p<0.05) direct effects, as shown by standardized regression coefficients, for i) HAZ were maternal height (0.38), being white (‐0.10), of high birth order (‐0.16), and having a helminth infection (‐0.08); and for ii) BMIZ were maternal weight (0.21), haemoglobin (0.14), and extremely low SES (‐0.19). Significant (p<0.05) indirect effects for i) HAZ were sex (‐0.02) and serum Se (‐0.02); and for ii) BMIZ was having a helminth infection (‐0.003). One of the most complex indirect pathways influencing both stature and ponderal growth involved helminths, which acted on serum Se and then serum Zn. In conclusion, multiple factors influence linear growth and body mass indices of these preschoolers, of which treatment of helminth infections with deworming is a modifiable risk factor with the potential to enhance somatic growth.

Grant Funding Source: Funded by University of Otago Research Fund and Nutrition Department PBRF fund
Evaluating a promotor‐led, nutrition and physical activity community intervention (624.4)
Authors: Friedman Maya , Jara Eddy , Fajardo Alexander
Published: June 2021

With Hispanic Americans 1.2 times more likely to be obese and 1.8 times more likely to be diagnosed with diabetes compared to non‐Hispanic Whites, El Sol Neighborhood Education Center and Loma Linda University School of Public Health developed a project to promote healthy eating and physical activity among Spanish‐speaking Latino homemakers and their families. This community‐based project utilized a promotor (i.e., community health worker) approach to implement six session curriculum (1.5 hours per session). A total of 600 Spanish‐speaking Latinas participated in the series. The behavioral objective was for participants to experience a change in key indicators of capacity to eat healthier and be more physically active (i.e., perceived benefits, knowledge, self‐efficacy, intention and behavior). Evaluation methods included a pre‐and post‐test design and administration of modified Photovoice tool that asked participants to draw what they felt impacted them the most from participating in the curricular sessions. Pre‐post community participant survey responses revealed significant increases (p<0.05) in: 13 out of the 18 knowledge questions and all nine of the behavioral intention questions (e.g., to exercise more; decrease sugary drinks; eat more fruits and vegetables, prepare healthy snacks for their children and decrease the frequency of eating out at fast‐food restaurants). The Photovoice drawings corroborated the survey results, highlighting positive changes in knowledge, intention, motivation and behavior. Findings suggest that that the promotor‐led nutrition and physical activity curriculum promotes healthier eating and physical activity among Spanish‐speaking Latinas. The authors recommend implementing this intervention with non‐Latino communities to see if these results can be validated and reproduced.

Grant Funding Source: Supported by Desert Health Care District and Loma Linda University Office of Public Health Practice
Impact of a course‐based nutrition intervention on health‐related behavior change based on the transtheoretical model (626.13)
Authors: Fernandes Nicolle
Published: June 2021

Readiness to change is a key factor in successful behavior change. The purpose of this study was to evaluate the impact of a course‐based nutrition intervention on behavior change in college students and to identify factors that promote/obstruct positive health behaviors. Participant‐categorized stages of change were identified to test the impact of the intervention to induce change. Of the 53 participants, majority were female (93%) and Caucasian (89%), and had an average age of 20 (18‐25) and body mass index (BMI) of 24.5 (19.1‐39.1). Health behaviors that students were most likely to engage in during and after the intervention were exercise (35%), increasing intake of fruits and vegetables (33%), whole grains (19%) and calcium (13%). Interestingly, the highest transformation according to the stages of change i.e. from pre‐contemplation or contemplation to preparation, action or maintenance was observed for reducing sodium intake (45%). This was followed by increasing calcium intake (32%), reducing fat (26%) and increasing fruits and vegetables (19%). While 22% of students indicated course attendance as a factor that aided success in behavior change, 43% indicated motivation (personal and external) to be the highest contributor to their success. Time seemed to be the most common barrier as indicated by 60% of participants. Although participants were predominantly health‐majors, only 39% identified health as a priority value in the pretest (39%). This increased to 56% posttest. In conclusion, course‐based nutrition education may be an effective approach in impacting students to adopt health behaviors based on their readiness to change.

Grant Funding Source: Scholarship of Teaching and Learning (SoTL), Ball State Univeristy
Neuroprotective effects of cheonggukjang prepared with B. amyloliquefaciens MJ1‐4 and Bacillus sp EMD17 (629.12)
Authors: Woo Ji Eun , Seo Ji Yeon , Kang Hye Ryung , Kim Seong Soon , Zakharova Alexandra , Kim Mi Hye , Kim Bo Ram , Han Mi Kyung , Kim Jeong Hwan , Kim Jong‐Sang
Published: June 2021

The health benefits of cheonggukjang are widely accepted. However, the sensory property of cheonggukjang such as unique odor is still a challenging issue which hampers extensive consumption.

In this study, we investigated the neuroprotective activity of cheonggukjang prepared with B. amyloliquefaciens MJ1‐4 (MJ1‐4) and Bacillus sp. EMD17(EMD17) which have better odor. Cheonggukjang was made with MJ1‐4 alone or combination with MJ1‐4 and EMD17 as well as with different incubation times. Aqueous ethanol (80%, v/v) extracts of each cheonggukjang prepared with MJ1‐4 alone and combination with MJ1‐4 and EMD17 were subjected to determination of neuroprotective activities, using MTT cytotoxicity and 2´,7´‐dichlorofluorescein diacetate (DCF‐DA) assays. The results showed that cheonggukjang prepared with MJ1‐4 alone and combination with MJ1‐4 and EMD17 had relatively high protective effect against glutamate‐induced cytotoxicity, decreased ROS accumulation, and caused higher NQO1 induction, compared to normal cheonggukjang in mouse hippocampal HT22 cells.

Grant Funding Source: Supported by NRF‐2010‐0027204 and 2011‐0009782 and IPET (High Value‐added Food TechnologyDevelopment Program, 2012, 112066‐3), S. Korea
Improvement of memory and cognitive function by alantolactone and isoalantolactone in mouse model (629.9)
Authors: Seo Ji Yeon , Kim Seong Soon , Kang Hye Ryung , Kim Bo Ram , Zakharova Alexandra , Woo Ji Eun , Kim Mi Hye , Han Mi Kyung , Kim Jong‐Sang
Published: June 2021

Our previous study demonstrated that alantolactone and its isoform isolated from Inula helenium had neuroprotective and antioxidative effects in primary cortical neurons. This study was conducted to assess the preventive effect of both sesquiterpenes from the impairment of memory and cognitive function in scopolamine‐induced mice. ICR mice (5 week‐old, male) were injected i.p. with 1 mg/kg body weight of scopolamine to induce amnesia. The memory and spatial learning ability of mice were evaluated by passive avoidance test, Y‐maze test, and Morris water maze test. We found that these two sesquiterpenes improved the cognitive function of mice injected with scopolamine in a dose‐dependent manner. In particular, the group treated with 1 mg isoalantolactone per kg body weight showed the longer latency time than positive control group. Moreover, isoalantolactone reversed the scopolamine‐induced spatial memory impairment of mice as assessed by Y‐maze test. The inhibition of acetylcholine esterase in hippocampus and cerebral cortex was postulated as the potential mechanism of the memory improvement by the compound. In conclusion, these results suggest that sesquiterpenes attenuate scopolamine‐induced memory impairment maybe through enhancement of the cholinergic nervous system via acetylcholinesterase inhibition.

Grant Funding Source: Supported by NRF‐2010‐0027204 and 2011‐0009782 and IPET (High Value‐added Food TechnologyDevelopment Program, 2012, 112066‐3), S. Korea
Opportunities and challenges for intersectoral convergence in the delivery of nutrition interventions in India (632.16)
Authors: Avula Rasmi , Kim Sunny , Ved Rajani , Pradhan Mamata , Menon Purnima
Published: June 2021

Background: In India, delivery of interventions to improve child nutrition requires active IC between the Ministries of Health and Family Welfare (MOHFW) and Women and Child Development (MWCD). This in turn requires linked actions in policy formulation, implementation, and monitoring & evaluation (M&E), but little is known about IC or how to optimize it.

Objectives: To identify opportunities and challenges to achieving IC in policy, implementation and M&E for nutrition at the national level and in 2 states.

Methods: Content analysis of policy & program documents, stakeholder interviews (n=38 national; 26 state).

Results: Policy: Some shared objectives, an articulated need for IC, and diverse task forces support convergence in policy. Nutrition interventions are part of MOHFW programs, but nutrition is not a core objective. Implementation: Operational designs recognize need for coordinated frontline service delivery; delivery structures provide IC opportunities. However, personal relationships and shared understanding of work needs, rather than formal mechanisms facilitated IC. At state & district levels, facilitators were political commitment, will and leadership. M&E: Lack of common indicators and monitoring systems limit IC.

Conclusions: There is potential to optimize IC but strong leadership, explicit roles and linked M&E systems could facilitate better IC for nutrition impact in India.

Grant Funding Source: Supported by: Bill & Melinda Gates Foundation, through POSHAN , led by IFPRI
Daidzein enhances efferocytosis via transglutaminase 2 and augmentation of Rac1 activity (639.10)
Authors: Yen Jia‐Hau , Chen Hsiao‐Ling , Tsay Gregory
Published: June 2021

Objective: Defective efferocytosis is cumulatively recognized in autoimmune and chronic inflammatory diseases. Based on our previous finding, an ethanolic extract from Glycine tomentella Hayata (GTH) can enhance mouse macrophage Raw264.7 efferocytosis (clearance of apoptotic cells). We have previously demonstrated that the major components of GTH are daidzein, catechin, epicatechin and naringin. Here, we explore the potential of each component for modulating efferocytosis activity.

Methods and results: For detecting the effect of daidzein, catechin, epicatechin and naringin on efferocytosis, RAW264.7 cells were cultured with CFDA‐stained apoptotic cells and assayed by flow cytometry. We found that daidzein is the main component of GTH, and it can enhance Raw264.7 efferocytosis dose‐dependently. Moreover, the increased effect of daidzein on macrophage efferocytosis activity is accompanied by TG2 at both the mRNA and protein levels. Conversely, TG2 knockdown attenuated the effect of daidzein on increased macrophage efferocytosis activity. After treatment with daidzein, increased phosphorylation was observed in Erk, but not P38 and JNK. Finally, we report that after daidzein treatment, Raw264.7 macrophages markedly increased Rac1 activity and decreased the mitochondrial membrane potential, which may contribute to efferocytosis.

Conclusion:Taken together, these data suggest that daidzein enhancement of macrophage efferocytosis activity was mainly through up‐regulation of TG2 expression and Rac1 activity. Daidzein may have the potential for treatment strategies for inflammatory diseases.

Grant Funding Source: Supported by NSC 99‐2811‐B‐040‐005, Taiwan
The proportion of stressed individuals who, on any given day, do not have a cold/flu may be increased by the intake of bifidobacteria: a randomized, double‐blind, placebo‐controlled study (639.13)
Authors: Langkamp‐Henken Bobbi , Rowe Cassie , Ford Amanda , Christman Mary , Dahl Wendy
Published: June 2021

Psychological stress is associated with cold/flu. This study determined whether probiotics increased the proportion of academically stressed students who, on any given day, did not have a cold/flu (ie, healthy day). Before, during, and after final exams (6‐wk period), students (20±2 y) took a probiotic (L. helveticus R0052, B. bifidum R0071, or B. longum ssp. infantis R0033) or placebo (n=142 to 147/grp) and self‐reported level of stress and symptom intensity (SI, 0=not experiencing to 3=severe) of 9 cold/flu symptoms. SI scores were summed; a score 蠄6 was counted as a healthy day. The effects of intervention, week, gender, sleep, stress, and categorized baseline SI scores (to control for individual variation), and 2‐way interactions were in the generalized linear mixed model. There was an effect of gender and an interaction between the intervention and stress, sleep, and SI category on the proportion of “healthy” individuals on any given day. B. longum showed a positive relationship with hrs of sleep on likelihood of reporting a healthy day. Pairwise comparisons (averaged over other effects) indicated that B. bifidum (0.99 ± 0.003) and B. longum (0.99 ± 0.003) differed from the placebo (0.97 ± 0.005) in the proportion of healthy individuals but L. helveticus (0.98 ± 0.004) did not. The data suggest that the proportion of stressed individuals who, on any given day, do not have a cold/flu may be increased by Bifidobacteria.

Grant Funding Source: Lallemand Health Solutions
Relationship between oxidative stress biomarkers in urine and erythrocytes, and to estimates of fecal Giardia intestinalis intensity in Guatemalan preschool children with a common menu offering (639.3)
Authors: Soto‐Méndez María , Aguilera Concepción , Mesa María , Campaña‐Martín Laura , Martín‐Laguna Victoria , Solomons Noel , Schümann Klaus , Gil Angel
Published: June 2021

Objectives: To describe urine oxidative stress biomarkers and erythrocyte antioxidant enzymes activity, and explore any association with prevalence or intensity of Giardia intestinalis infection (Gi) in preschoolers attending 3 government‐subsidized daycare centers in the Guatemalan Western Highlands Methods: 74 24‐h urine, blood and fecal samples were collected the same week, among 36 F and 38 M to measure oxidative damage to DNA (8‐hydroxydeoxyguanosine {8‐OHdG}) and lipid (15‐Isorpostane F2t {F2‐Iso}), the erythrocyte activity of Catalase (CAT) and Superoxide Dismutase (SOD) Glutathione Reductase (GSHR) and Glutathione Peroxidase (GSHPx). Gi prevalence and intensity were assessed by ELISA kit procedures Results: CAT activity correlates significantly with 8OHdG (r=0.242, p=0.038) and F2‐Iso (r=0.468, p<0.001), whereas SOD activity correlates only with F2‐Iso (r=0.256, p=0.028). Positive correlations were also found between Gi intensity and CAT (r=0.337, p=0.003) and F2‐Iso (r=0.291, p=0.012). Median values of rbc CAT activity (p=0.016) and urine F2‐Iso (p=0.023) differed between children who were Gi+ and Gi‐.

Conclusion: We find heretofore unrecognized associations between rbc and urine oxidative stress biomarkers and with the prevalence and intensity of Gi in our study population Funded by: Fundación Iberoamericana de Nutrición (FINUT), Spain and The Hildegard Grunow Foundation (HGF), Germany

Grant Funding Source: Supported by Fundación Iberoamericana de Nutrición (FINUT) and The Hildegard Grunow Foundation (HGF)
1,25‐Dihydroxyvitamin D alters the pentose phosphate pathway in Harvey‐ras oncogene transfected MCF10A breast epithelial cells (644.3)
Authors: Wilmanski Tomasz , Zheng Wei , Donkin Shawn , Raftery Daniel , Bequette Brian , Burgess John , Teegarden Dorothy
Published: June 2021

The pentose phosphate pathway (PPP) has emerged as a mediator of cell proliferation and redox balance with its up‐regulation observed in early stages of cancer progression. Here we demonstrate that the active vitamin D metabolite, 1,25‐dihydroxyvitamin D (1,25D), alters the PPP in untransformed (MCF10A) and Harvey‐ras oncogene transformed (MCF10A‐ras) breast epithelial cells. Following four day treatment with 10 nM 1,25D, [13C6] glucose flux into the PPP was decreased by 23% in MCF10A and 16% in MCF10A‐ras cells compared to vehicle. Additionally, 1,25D reduced the abundance of nucleosides measured by NMR by 24% and 25% in MCF10A and MCF10A‐ras cells, respectively. There was no difference following 1,25D treatment in mRNA expression of transaldolase, an enzyme in the nonoxidative branch of the PPP. Interestingly, mRNA levels of the rate limiting enzyme glucose‐6‐phosphate dehydrogenase (G6PD) were increased 2.3±0.31 fold in MCF10A and 2.18±0.17 fold in MCF10A‐ras cells compared to vehicle. G6PD is a central antioxidative enzyme; however its up‐regulation correlated with a paradoxical 50% increase in reactive oxygen species, as measured by the fluorescent probe dichlorofluorescein diacetate. Together these results suggest 1,25D inhibits glycolytic flux into the PPP, thereby potentially decreasing the reducing potential and biosynthetic substrates necessary for proliferation.

Grant Funding Source: Supported by NIH R25CA128770 and NCI CCSG CA23168
Effects of processing, cooking, and storage on β‐carotene retention and bioaccessibility in biofortified cassava (Manihot esculenta) (646.4)
Authors: La Frano Michael , Zhu Chenghao , Burri Betty
Published: June 2021

Biofortification of cassava with β‐carotene is currently being tested in African populations where cassava is a staple food and vitamin A deficiency is a public health problem. Measuring the impact of traditional African processing and cooking on β‐carotene concentration and bioaccessibility of cassava‐based foods can help direct plant breeding efforts and estimate the potential impact of biofortified cassava on vitamin A status. Biofortified cassava was processed and cooked to prepare the following traditional dishes: gari, eba, fermented and non‐fermented fufu, and boiled cassava. Eba, non‐fermented fufu, and gari retained the most β‐carotene (43.6, 42.2, and 41.0%, respectively) while boiled cassava and fermented fufu retained the least (24.9, and 21.7%, respectively). Gari and boiled cassava bioaccessibility was 17‐ and 12‐fold higher than raw cassava, respectively (P < 0.001), while increases in bioaccessible β‐carotene concentrations in eba, and fufu products were not significant. Unfortunately, after just three days of storage at room temperature fermented flour and gari preparations lost >62% β‐carotene, while nonfermented flour was more stable. These results suggest that biofortified cassava prepared as gari or boiled might provide useful amounts of β‐carotene to alleviate vitamin A deficiency, if storage conditions are optimized.

Grant Funding Source: Supported by HarvestPlus
Anti‐oxidative characteristics of Korean marine algal extracts (647.15)
Authors: Lee Je‐Hyuk
Published: June 2021

Among 51 marine algae distributed in Korean coast, Sargassum muticum (Yendo) Fensholt, Ishige okamurae Yendo, Ecklonia cava Kjellman, Polysiphonia morrowii Harvey, Dictyopteris undulata Holmes, Sargassm micracanthum (Kutzing) Endlicher, and Sargassum macrocarpum C. Agarch, were investigated to have the potent antioxidant activity. Additionally the reducing power of marine algae‐extracts was totally consistent with DPPH radical scavenging activity. Because little is known about Sargassum muticum (Yendo) Fensholt, Ishige okamurae Yendo, Ecklonia cava Kjellman, Polysiphonia morrowii Harvey, Dictyopteris undulata Holmes, Sargassm micracanthum (Kutzing) Endlicher, and Sargassum macrocarpum C. Agarch, these marine algae may act as the candidates for potent antioxidant food materials, and are expected to have the functional activity for foods and pharmaceuticals for oxidative diseases.
Boron content of California raisins (647.2)
Authors: Carughi Arianna , Mike Hurley
Published: June 2021

Both epidemiological and laboratory evidence suggest a role for boron in various physiological processes and in human health. While boron is essential in plants and other organisms it is suspected to be essential in humans but this has not been directly proven. As an element ubiquitous in the earth’s crust, boron is a natural constituent in the human diet. Fruits, vegetables, nuts and pulses are natural sources of boron in the diet the level reflect the soil concentrations where these crops are grown. Raisins have been reported as among fruits with the highest boron content. This study measured boron content of sun‐dried raisins from grapes grown in four distinct sites in the San Joaquin Valley. Consistent with previous reports, values ranged from 1,373 µgrams/100 grams to 2,245 µgrams/100 grams fresh weight, placing raisin among top boron contributors to the American diet.
Whole muscadine grape components increase phenolic content and enhance bioactive compounds of fruit smoothies during processing and storage (647.34)
Authors: Stallings Kendra , Harris Gabriel
Published: June 2021

Whole fruit purees, including seeds and skins, may increase the bioactive properties of fruit smoothies, an increasing market. The objectives of this study were to determine if adding whole muscadine puree (MP) to smoothies enhanced nutritional properties and protected labile compounds through thermal processing and storage. Smoothie formulations, and individual components, were compared with and without MP. Samples were thermally processed (71°C, 10 s) then refrigerated. Total antioxidants, phenolics, anthocyanins, and ascorbic acid were analyzed on the fresh, processed, and refrigerated samples. Higher than expected antioxidant values were seen in smoothies based on results from individual ingredients suggesting a synergistic effect enhancing bioactives when adding whole fruit components. Antioxidants increased 16% and 6% in smoothies during processing with and without MP, respectively. Phenolics, ascorbic acid, and anthocyanins were retained 2.1%, 0.2%, and 8.0% more, respectively, during processing in smoothies with MP compared to smoothies without MP. Smoothies containing MP retained 13% more phenolics and 4% more anthocyanins during storage compared to smoothies without MP. During storage, antioxidants increased 11% with MP and decreased 5% without MP. A protective effect was seen from the natural preservative properties of MP on bioactives in smoothies during processing and storage.
Chemical properties and antioxidant activity of beverage with ethanol extracts from the roasted Gugija (Lucii fructus) (647.41)
Authors: Lee Kyongae , Park Jinsook
Published: June 2021

The antioxidant activity of beverage with ethanol‐extract from the roasted Gugija (Lucii fructus) were studied along with free amino acid and mineral composition analysis. Antioxidant activity determined by DPPH and ABTS radical scavenging activity. Five major free amino acids of the Gugija beverage were methionine (20.91 mg%), aspartic acid (16.91), alanine (12.52 mg%), %), proline (11.39 mg%), and arginine (7.53 mg%). The three major mineral in beverage with the ethanol‐extracts of the roasted Gugija were potassium, magnesium and calcium at 472.9, 16.47 and 9.66 mg% , respectively. Electron donating ability and ABTS radical scavenging activity of the Gugija beverage were 94.8 and 94.9%, respectively, of control L‐ascorbic acid, whereas those of ethanol‐extract were 85.4%, and 80.4%, respectively. Contents of total polyphenols and flavonoid of Gugija beverage were 325.5 mg tannic acid/g and 97.9 mg quercetin /g, respectively. The total polyphenol content and flavonoid was higher in Gugija beverage, which correspond to 3.77 and 4.49 times, respectively of that of ethanol extract. High DPPH and ABTS radical scavenging activity of the Gugija beverage were possibly related with amino acid composition as well as high contents of polyphenols and flavonoids in beverage
Soybean cake as a source of stable bioactive compounds (647.43)
Authors: Silva Fabrício , Silva Cecília , Torres Alexandre , Teodoro Anderson , Monteiro Mariana , Perrone Daniel
Published: June 2021

Soybean cake is a byproduct of soybean oil processing rich in bioactive compounds. In this study we evaluated the effect of soybean cake extracts on cell viability, cycle and apoptosis in human colon adenocarcinoma cell line (HT‐29) and their stability under different storage conditions. Soybean cake from soybean oil industry (SCI) and produced under laboratory conditions (SCL) were extracted with a ternary mixture (water:ethanol:ethyl acetate), lyophilized and incubated with HT‐29 cells at concentrations (10 to 400 mg/g) from 6 to 96 h. The dry extracts were stored in freezer or room temperature (light or dark) and the following analysis were conducted at regular intervals during 120 days: spectrophotometric assays for phenolic compounds, flavonoids and saponins; antioxidant capacity (AC) by FRAP and TEAC assays; isoflavones and soyasaponins by LC‐DAD‐MS. SCL and SCI extracts showed high levels of phenolic compounds (12.36 and 10.45 mg gallic acid/g), flavonoids (7.24 and 9.98 mg genistein/g), saponins (224.86 and 253.99 mg/g) and AC (FRAP: 71.35 and 91.57 µmol Fe+2/g; TEAC: 105.02 and 103.90 µmol Trolox/g). LC‐DAD‐MS analysis revealed that SCI extract presented higher levels of β‐glycosylated isoflavones (3.67 mg/g) than SCL (0.72 mg/g). Aglycone isoflavones were present in higher levels in SCL extract (3.16 mg/g) than in SCI (0.82 mg/g). Both extracts reduced in a time‐dose dependent fashion cell viability when compared to control at concentrations starting from 100 mg/g. These reductions ranged from 4% to 84% and from 24% to 85% when cells were incubated with SCL and SCI, respectively. After incubation with SCL a decrease in the percentage of cells at G0/G1 phase and an increase in G2/M phase was observed, as well as an increase in the apoptosis rate. After 120 days of storage, bioactive compounds’ levels did not change, with the exception of FRAP values. Extracts showed anticancer effects in vitro and soybean cake is a potential source of stable bioactive compounds for in vivo studies.

Grant Funding Source: FAPERJ, CNPq, CAPES, UFRJ