bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of postprandial hypoglycemic effect of Ecklonia cava extract in healthy subjects using three different challenge models
Authors: Kim Yunsoo , Lee Hyo Jin , Ahn Young sook , Kwon Oran
Published: November 2023

Repetitive postprandial fluctuation in glucose concentration evokes significant increases in risks of cardiovascular disease, visceral adiposity, and insulin resistance. Ecklonia cava (seapolynol) is a brown alga that is abundant in Jeju Island, Korea. It contains a variety of phloroglucinol‐based polyphenols, as phlorotannins that have inhibitory activities against α—amylase and α‐glucosidase in vitro. According to a recent publication, Ecklonia cava extract (EC) exhibited a peak plasma concentration of bioactive compounds and their metabolites more than 2 hours after oral ingestion. Based on this background information, we intended to compare the postprandial hypoglycemic effects of EC in healthy subjects by performing three independent randomized controlled crossover trials. The first study was performed in a high‐carbohydrate meal challenge (four slices of white bread with strawberry jam). Blood responses were determined at 0, 15, 30, 60, 120 and 180 min. The second study was performed using a high fat and carbohydrate formula (60g fat and 75g maltose). The blood samples were obtained at the same time intervals with the first study. The last study was conducted by providing two consecutive maltose loadings (75 g for each) with a 2‐hour interval. Blood samples were obtained at 0, 15, 30, 60, 120, 135, 150, 180 and 240 min for determination of glucose and insulin levels. The EC administration showed a significant reduction in postprandial glucose and insulin levels during the first 30 min when compared with the placebo group; and in postprandial triglyceride level at 180 min. The most effective dose was determined at 360 mg. The high‐fat and carbohydrate formula showed a less sensitive phenotype compared with either a high‐carbohydrate meal challenge or a maltose loading.

Support or Funding Information

This work was supported by the Bio‐Synergy Research Project (NRF‐2012M3A9C4048761) from the Ministry of Science, ICT, and Future Planning, and the BK21PLUS (Brain Korea 21 plus) program (22a20130012143) from the Ministry of Education, Republic of Korea
Melatonin improves insulin resistance and hepatic steatosis through attenuation of ER stress
Authors: Heo Jeein
Published: November 2023

Non‐alcoholic fatty liver disease (NAFLD) is increasingly recognized as one of the most common liver diseases. However, there is no specific way to treat or prevent it. Melatonin is a key molecule to regulate body rhythms. It also acts as a potent anti‐oxidant, and the melatonin deficiency is associated with various diseases, including diabetes and obesity; however, the action mechanism is not well known. In addition, fetuin‐A is one of the hepatokines and is known to be associated with insulin resistance and type 2 diabetes. However, it is not known whether melatonin can improve hepatic insulin resistance by down‐regulation of fetuin‐A expression.

We used HepG2 cells as a model to study whether melatonin can reduce palmitic acid‐induced insulin resistance. In HepG2 cells treated with (300 uM) of palmitic acid, phosphorylated AKT expression was decreased and fetuin‐A expression was increased, but this effect was inhibited by the treatment of 10 uM of melatonin. Moreover, melatonin decreased palmitic acid‐induced ER stress markers, CHOP, Bip, ATF‐6, XBP‐1, ATF‐4, IRE‐1, and PERK. In addition, thapsigargin, an ER stress activator, increased the expression of fetuin‐A, and ER stress markers, and it was inhibited by melatonin treatment.

In addition, in the high fat diet (HFD) mice, oral treatment of 100 mg/kg/day melatonin for 10 weeks decreased body weight, epididymal fat mass, and hepatic steatosis. Insulin sensitivity and glucose tolerance was improved and plasma insulin was decreased by melatonin in HFD mice. Fetuin‐A expression in liver and serum of HFD mice was decreased by melatonin treatment. ER stress markers in liver of HFD mice was decreased by melatonin.

Therefore, we suggest that melatonin can improve hepatic insulin resistance and hepatic steatosis, and this may be due to the reduction of ER stress and resultant fetuin‐A expression.

Support or Funding Information

This work was supported by Research Grant (2015R1C1A2A010520) of National Research Foundation of Korea.
Cytokine Production by Insulin Resistant Adipocytes
Authors: Levenberg Kate Robbins , Laakko‐Train Tonya
Published: November 2023

The pathogenesis of type II diabetes is associated with obesity‐induced chronic inflammation. This association is thought to be mediated by pro‐inflammatory cytokines that inhibit insulins' signaling pathways. This study aimed to further the knowledge on the development of type II diabetes by identifying the cytokines that are expressed by adipocytes in an insulin resistant state. In order to do this, the 3T3‐L1 fibroblast cell line was first differentiated into adipocytes by exposure to dexamethasone (0.25 pM) and methylisobutylxanthine (0.5 mM) for two days, followed by exposure to insulin (2 pg/ml) for two days. Differentiation was evaluated by lipid droplet accumulation. The differentiated adipocytes were estimated to contain an average of 40 lipid droplets with a diameter of 7–15 μm each, as compared to an average of 4 droplets, with a diameter of 2–3 μm, in each fibroblast cell. Insulin resistance was induced by exposure to insulin for a minimum of 8 hours at a concentration of 10 nM. The secretion of 32 different cytokines by insulin resistant and non‐insulin resistant adipocytes was then evaluated by a membrane‐based antibody array. It was found that RANTES, TNF‐r1, TIMP‐1, and MCP1 were significantly upregulated in an insulin resistant state (p<.05). This work indicates that these specific cytokines may interfere with insulins' signaling pathway. Future research should study the interaction between these cytokines and the insulin signaling pathway, to further understand the link between inflammation and type II diabetes.
The role of the subunits of the Conserved Oligomeric Golgi Complex in filamentous growth of the fungus Aspergillus nidulans
Authors: Gremillion Sara
Published: November 2023

Filamentous fungi grow as thin, highly‐branched cells with new, polarized growth occurring at the tips. Growth is maintained by the continuous export of plasma membrane and cell wall materials via vesicles from the Golgi apparatus. In order to maintain proper Golgi function, this organelle must continually recycle membrane components and enzymes lost by departing vesicles. The Conserved Oligomeric Golgi (COG) complex is a tethering complex involved in the docking of retrograde vesicles within the Golgi. Using the filamentous fungus Aspergillus nidulans, we are studying the eight subunits of the COG complex, COG1–8. Through a promoter replacement strategy, we have created new strains of A. nidulans that express COG proteins at lower‐than‐normal levels in order to determine which subunits are important for the function of this complex, and therefore, important for growth. When expression of the subunits COG1–4 is reduced, fungal cells are unable to maintain polarized growth. Alternatively, under‐expression of subunits COG5–8 does not appear to have a negative effect on filamentous growth.

Support or Funding Information

This research is supported by internal grants provided by Armstrong State University.
Cellular Proteases Prime Extracellular Vesicles for Intercellular Communication
Authors: Qing Enya , Hantak Michael , Gallagher Tom
Published: November 2023

Extracellular vesicles (EVs) can mediate intercellular communication by encasing cytoplasmic cargo and transferring this luminal material into nearby cells. This EV‐mediated transfer process is important for maintaining metazoan homeostasis (a positive effect) and is also implicated in metastatic tumor progression (a negative effect). To transfer cytoplasmic cargo, an EV must fuse its membrane with the plasma or endosomal cell membrane on a target cell. A poor understanding of this fusion process has hindered EV biology.

We and others are impressed by the similarities between EVs and enveloped viruses. During enveloped virus infection, luminal cargos are transferred through an analogous fusion process. Activating viral membrane fusion frequently requires host proteases. Therefore, we hypothesized that cellular proteases prime EV‐mediated cargo transfers. We quantified EV cargo transfers with a membrane fusion‐dependent reporter complementation assay, where signals are generated only after EV‐mediated delivery of reporter (luciferase) proteins. We found that transmembrane protease serine 2 (TMPRSS2), a protease known to activate several viral fusion‐catalyzing enyzmes, profoundly increased EV‐mediated cargo transfers. TMPRSS2 mediated its effects within the cells producing EVs, suggesting the existence of a protease‐primed, virus‐like fusogen that traffics on EVs to prompt EV‐target cell coalescence and cargo delivery. Our findings may offer increased understanding of EVs and their roles in regulating crucial biological processes, and may provide tools to manipulate EVs for therapeutic purposes.
Comparison of the Structure Between the Wild Type Lamin A/C Protein and the Cardiac Disease Causing Variant D192G
Authors: Wall Susan , Akman Alexandra , Chow Alyssa , Deng Mirella , Dumitrascu Flavia , Jiang Max , McCartney Maggie , Nguyen Tanya , Rulko Theo , Sullivan Nicholas , Wang Lauren , Xiao Mason , Nicolas Hannah
Published: November 2023

The lamin A/C (LMNA) gene codes for A type lamins which are key components of the nuclear lamina and are involved in maintaining the structure of the nucleus and its processes. Mutations in LMNA cause a group of diseases known as laminopathies. For this project, the lamin A/C variant affecting amino acid 192 is studied. The causal mutation results in an amino acid change from glycine [G] (smaller, non‐polar, uncharged) to aspartic acid [D] (larger, polar, charged). This D192G variant is linked to a severe form of a cardiac disease called Dilated Cardiomyopathy (DCM), where cardiomyocytes are presented with multiple nuclear abnormalities. In vitro experiments with the D192G variant showed abnormal nuclear localization of Protein Kinase C alpha (PKC‐α). PKC‐α is the major protein kinase C isoform found in the heart and skeletal muscles. This enzyme phosphorylates a host of proteins and uses A‐type lamins to reach its nuclear targets. In order to help elucidate the potential role of PKC‐α as the link between the disease phenotype and lamin mutation, a structural comparison between the wild type lamin A and the D192G variant was performed in silico. Moreover, the PKC‐α binding site on the wild type and mutant lamin A were compared in order to determine if a conformation change has occurred. Since there is currently no full‐length crystal structure for either the wild type or D192G lamin A/C variant, the I‐TASSER software suite was used to generate simulated protein structures. The top five structures for the WT and variant simulations were examined. The final models selected to most likely represent reality were chosen based on various factors such as computed C‐score, what is known in literature and human analysis of the simulations. The results show that the D192G variant predominantly takes a globular form which is very comparable to the structure of the wild type lamin A found in the nucleoplasm. This globular form also potentially explains the lamin aggregates found in cells expressing the D192G variant. However, this globular variant conformation significantly differs when compared to the known and prevalent wild type lamin A conformation which is the globular head‐alpha helical rod‐globular tail structure. In particular, the conformation of PKCα binding site on lamin A is also significantly altered. The Ashbury College MSOE Center for BioMolecular Modeling SMART Team used 3‐D modeling and printing technology to examine the structure‐function relationships of the wild type lamin A/C protein and the D192G variant.
MELAS and POLG exhibit high rates of hypertension with differential resistance to anti‐hypertensive drugs suggesting divergent pathogenic mechanisms
Authors: Pauls Andrew David , Nevay Dayna‐Lynn , Young Dana , Sandhu Vikrant , Yeung Darwin F , Tsang Michael MY , Tsang Teresa SM , Mattman Andre , Mezei Michelle M , Poburko Damon
Published: November 2023

Background

We previously reported elevated hypertension (HT) rates in adults with the m.3243A>G mitochondrial encephalomyopathy, lactic acidosis, and stroke‐like episodes (MELAS) mutation. The cause of HT in MELAS is not known but could be linked to dysautonomia and endothelial dysfunction. Hypertension is also reported in transgenic mice that have an error‐prone gamma DNA polymerase (POLG) that increases mtDNA mutation rate. In contrast to MELAS, HT in POLG−/− mice appears to be due to hypertrophy of the tunica media. It is not known if humans with pathogenic POLG mutations similarly exhibit HT, and whether the hemodynamic basis of HT in MELAS and POLG are consistent with common or distinct pathogenic mechanisms.

Purpose

We aimed to examine whether POLG patients have an increased risk of HT, and whether the hemodynamics and therapeutic sensitivity of such HT resembles that of HT in MELAS patients. Specifically, we hypothesized that POLG patients would be prone to isolated systolic HT, while MELAS patients should exhibit remodeling of resistance vessels.

Methods

We conducted a chart review at the Adult Metabolic Disease Clinic, Vancouver General Hospital of 35 MELAS and 26 POLG patients in accordance with institutional human research ethics guidelines. Demographic factors, including age, weight, BMI, sex, smoking status and diabetes status were collected. Patients were scored as hypertensive if they were on anti‐hypertensive drugs or had consistently elevated blood pressure measurements in accordance with the Canadian Hypertension Education Program Guidelines.

Results

MELAS and POLG patients showed increased prevalence of HT (20–39 yr age group: 30% MELAS, 14% POLG; 40–59 yr: 71%, 57%; 60–79 yr: 83%, 81%) compared to the Canadian population. Hypertension was not associated with sex, smoking or BMI, and exceeded expected levels given 50% of MELAS patients were diabetic. In the 40–59 and 60–79 yr age groups, antihypertensive drugs controlled systolic blood pressure in MELAS but not in POLG patients (40–59 yr: POLG 145 ± 21 mmHg n = 8 vs MELAS 122 ± 23 n = 12; 60–79 yr: POLG 157 ± 6 n=4 vs MELAS 134 ± 21 n=5), while diastolic blood pressure did not differ between MELAS and POLG. Preliminary analyses suggest that MELAS, but not POLG, patients exhibit elevated peripheral resistance despite therapeutic correction of blood pressure.

Conclusions

MELAS and POLG increase the risk of HT, which can increase the propensity for cardiac hypertrophy and cerebrovascular problems. Signs of vascular remodeling as a contributor to HT suggest that early therapeutic treatment at the onset of pre‐hypertension may be warranted to prevent and control HT in these diseases, especially POLG. These findings warrant detailed hemodynamic assessments in larger populations.
Improving the Recruitment, Retention, and Success of Students in STEM Disciplines
Authors: Baker Dianne M , Slunt Kelli M
Published: November 2023

The University of Mary Washington, a public and primarily undergraduate liberal arts institution, is completing a five‐year STEM Talent Expansion through Research, Engagement, Preparation, and Scholarships (STEREPS) project. This NSF‐DUE funded project was designed to test and assess ways to enhance interest in STEM careers and student performance STEM education, with a focus on underrepresented student populations. To increase student interest in STEM, and to recruit underrepresented students to major in STEM fields at UMW, we implemented a summer STEM research mentoring program for rising high school seniors, and offered small scholarships to entering and continuing STEM majors. To improve student success in STEM courses, we implemented an enhanced summer bridge program, including a new interdisciplinary course (IDIS 101) aimed to develop skills necessary for STEM courses, and a peer‐assisted student study (PASS) program during the academic year to increase success in traditionally challenging gateway courses. Analysis to date indicates that the summer research program was effective in recruiting students to study STEM fields at UMW, and that PASS participation was associated with higher grades in the introductory chemistry courses. The assessment of the efficacy of the summer bridge program is underway. Our lessons and conclusions about best practices will be discussed.

Support or Funding Information

Support for this work was provided by the National Science Foundation's Science, Technology, Engineering, and Mathematics Talent Expansion Program (STEP) under Award No. 1068150.
Simulating a Graduate Student Experience in an Undergraduate Course
Authors: Bednarski April Elaine
Published: November 2023

In designing this new research‐based laboratory course for undergraduates, an important goal was to include as many elements of a beginning graduate student experience as possible. The research topic for this course is enzyme analysis. Students begin with structural data for an enzyme then propose a single amino acid mutation. As the semester unfolds, students design, synthesize, and analyze the mutant enzyme. During this time, students participate in group meetings, journal clubs, journal‐based lab reports, and a final poster session. Steps to create an interactive learning environment and facilitate bonds between students were taken to mimic the graduate student experience as well as foster collaboration between the students. Student survey data from three years has been compiled and analyzed. In addition, the course was assessed using the CURE survey and compared with SURE survey results. Initial results show students rank the experience similar to a traditional summer undergraduate research experience.
Dll4: “Notch” Your Normal Pathway
Authors: Barmantje Emily , Nelson Erica , Agrawal Aayush , Chanjeevaram Smrithi , Jhaveri Aditya , Lazar Michelle , Li Katrina , McManus Darby , Santebennur Nikhita , Schlidt Mia , Medhora Meetha
Published: November 2023

The rising threat of nuclear terrorism necessitates proactive research to mitigate injury by radiation. Since one of the most damaging effects of radiation is the reduction of small blood vessels in healthy tissue, radiation‐induced organ damage may be lessened by increasing blood vessel density. The Notch‐Dll4 pathway plays an integral role in the development of blood vessels (angiogenesis). More recently, it has been reported to modulate mammalian vessel regression by initiating vessel occlusion that ultimately leads to vessel death. Expressed on endothelial cells lining blood vessels and on certain bone‐marrow derived cells, Delta‐like ligand 4 (Dll4) is a Notch‐receptor ligand. Ser28, Leu107 and Pro206 are all critical amino acids in ligand interaction between Notch and Dll4. Dll4 regulates vascular development by modulating tip and stalk cells in maturing vessels. Moreover, genetic and pharmacological inhibition of Dll4/Notch prevents capillary regression in a model of retinopathy. These results suggest that Dll4 inhibition may reduce the development of many diseases stemming from vascular insufficiency including radiation injuries. The Brookfield East High School SMART (Students Modeling A Research Topic) Team has designed a model of Dll4 using 3D printing technology to investigate structure‐function relationships. Radiation is used clinically to treat cancer but healthy tissue surrounding a tumor invariably receives damaging doses. By understanding the role of Dll4 in vascular regression, excessive damage to normal tissues can be avoided during radiotherapy or from a nuclear accident or attack.
“Buzz Off” Zika
Authors: Grant Fran , Arendt N. , Ducheny K. , Norton M. , Ovsepyan T. , Ruege M. , Wildermuth H. , Silvaggi Nicholas
Published: November 2023

Zika virus, or ZIKV, a mosquito‐borne pathogen, is a current public health concern due to its link to microcephaly and Guillain‐Barré syndrome. A flavivirus, ZIKV contains an RNA genome translated using the host cell's protein synthesis machinery. The 5′ end of the viral RNA possesses a “cap” of methylated bases that protect it from being destroyed by the host's immune system. This cap is produced by a virally‐encoded methyltransferase enzyme, the NS5 methyltransferase (MTase). Since the 5′‐RNA cap is required for viral reproduction, inactivating the NS5 MTase with a small molecule inhibitor should be a viable treatment for ZIKV infections. The NS5 MTase transfers CH3 groups from the ubiquitous methyl donor molecule S‐adenosylmethionine (SAM) to specific positions on bases at the 5′ end of the viral RNA. Based on the binding mode of SAM observed in a crystal structure (5KQR), an analog of SAM was developed that could inhibit the enzymatic activity of the MTase. SAM binds in a pocket composed of Ser56, Arg84, Trp87, Thr104, Lys105, His110, Glu111, Asp131, Val132, Phe133, Asp146, and Ile147. The Grafton SMART (Students Modeling A Research Topic) Team has modeled the NS5 MTase with either SAM or the hypothetical inhibitor MO2 bound using 3D printing technology. Models with such compounds bound to the enzyme provide information that might help increase the potency of potential inhibitors and improve their selectivity for the ZIKV MTase.

Support or Funding Information

This work was supported by the National Institutes of of Health Science Education Partnership Award (NIH‐SEPA 1R25OD010505‐01) and the National Institutes of Health Clinical and Translational Science Award (NIH‐CTSA UL1RR031973).
RPE65 – Essential Visual Cycle Protein
Authors: Kaiser Carl , Ahn P. , Arnhold K. , Bartos S. , Burkle B. , Chang M. , Coury M. , Cowan N. , Dittrich N. , Foster R. , French J. , Gamblin T. , Gorski J. , Hameed M. , Homberg L. , Johnson R. , Khan S. , Lewis E. , Otten J. , Rivera M. , Sabatino A. , Sargent T. , Schimmels J. , Smith A. , Stegeman R. , Strom D. , Strom J. , Tsuji B. , Visaya C. , Yamat J. , Yorke N. , Klestinski Keith , Huckenpahler Alison
Published: November 2023

Leber congenital amaurosis (LCA) is a genetic disease associated with blindness that is caused by mutations in retinal pigment epithelium (RPE65), an enzyme involved in the visual cycle. When light hits photosensitive pigments in the retina, it conformationally changes 11‐cis‐retinol (a form of vitamin A) to 11‐trans‐retinol. This conversion allows for a series of chemical reactions and the formation of electrical signals that communicate with the brain. RPE65 is essential to the cycle because it changes 11‐trans‐retinol back into 11‐cis‐retinol‐‐recycling the enzyme and allowing for sustained vision[1] . The Marquette High SMART (Students Modeling A Research Topic) Team used 3D printing technology to model how mutations in LCA affect RPE65's structure and function. RPE65 has 533 amino acids folded into sheets that arrange themselves into seven propeller blades, which collectively create a mostly hydrophilic tertiary structure. One face of RPE65 is hydrophobic and is associated with the microsomal membrane. A hydrophobic, tunnel‐like structure leads to an iron cofactor that facilitates its enzymatic efficiency. The iron cofactor is held in the active site by the histidine residues H180, H241, H417, and H527 in a coordinated motif. Mutations near the active site cause lowered enzymatic activity and LCA. Other residues that are commonly mutated in LCA include Gly40, Arg91, Glu102, Arg124, His182, and Val473. Ocular imaging data has shown phenotypic changes in the photoreceptors in patients with RPE65 mutations. With novel gene delivery methods and other treatment strategies, these technologies will be useful to understanding how to treat LCA and other retinal diseases. [2]

Support or Funding Information

This work was supported by the National Institutes of of Health Science Education Partnership Award (NIH‐SEPA 1R25OD010505‐01) and the National Institutes of Health Clinical and Translational Science Award (NIH‐CTSA UL1RR031973).
Inhibition of δ‐Opioid Receptors
Authors: Krukar Paula , Burmeister Katrina , Davis Evan , Dragseth Luke , Gent Austin , Janssen Andrew , Koch Stephen , Shaheer Mohammad , Panta Shadika , Brown Katie , Cunningham Christopher W.
Published: November 2023

Opioid addiction and abuse affects between 26 and 36 million people worldwide (Volkow, 2014), prompting researchers to try to design a drug that both has painkilling effects but does not create a dependency. Most opioid analgesics work through activation of mu opioid receptors (MOR). They naturally bind to opioid peptides, such as endomorphins 1 and 2, in order to regulate pain. Another important target is the delta opioid receptor (DOR). Inhibition of DOR has been shown to slow the development of addiction to MOR agonists in animal models of pain. DOR are proteins that transpermeate the cell membrane with seven alpha helices. Schiller and colleagues first reported an endomorphin analogue, DIPP‐NH2, that activates MOR and also inhibits DOR. DIPP‐NH2 is structurally similar to the natural endomorphins, so it can bind DOR through similar mechanisms. The DIPP‐NH2 interacts with the Met132, Tyr129, Val217, Ile277, and Trp284 on DOR. Moreover, one of the two methyl groups on DIPP‐NH2 engages Val281 and Ile277. Finally, a salt bridge forms between the DIPP‐NH2 N‐terminus and Asp128, and this salt bridge is crucial for opioid receptor ligand recognition. The Whitefish Bay High School SMART team (Students Modeling a Research Topic) is modeling DOR using 3D printing technology. DIPP‐NH2 may be the basis of a non‐addictive painkiller, which would be a significant advancement toward alleviating the major societal and financial problem of opioid addiction.

Support or Funding Information

This work was supported by the National Institutes of of Health Science Education Partnership Award (NIH‐SEPA 1R25OD010505‐01) and the National Institutes of Health Clinical and Translational Science Award (NIH‐CTSA UL1RR031973).
NMDAR Helps You Think, So Please Reconsider That Extra Drink
Authors: Sampe David , Block Gavin , Davis TJ , Dong Nancy , Freuler Jonah , Mietkowski Mitchell , Rowney Toni Jo , Stoehr Noel , Peoples Robert W.
Published: November 2023

According to the U.S. Centers for Disease Control and Prevention, each year 88,000 people die from alcohol‐related causes in the United States. Ethanol misuse, a deadly and expensive societal problem, cost the United States $249 billion in 2010 (1). Ethanol targets many proteins in the brain. Specifically, it acts as an inhibitor of the N‐Methyl‐D‐aspartate receptor (NMDAR), which mediates much of the excitatory synaptic transmission in the brain. Normally, the NMDAR is activated when glutamic acid binds to it allowing positive ions to flow through the cell membrane; this facilitates learning and memory in the brain. However, ethanol inhibits the activity of the NMDAR, interfering with synaptic transmission. A type of glutamate ion channel receptor found in neurons, NMDAR, has four domains: the amino terminal domain, the ligand binding domain, the membrane‐associated (M) domains, and the carboxy terminal domain. NMDAR is a heterotetrameric protein with two GluN1 and two GluN2 subunits. Ethanol interacts with NMDAR in the M domains with specific amino acids including Gly638 in the GluN1 subunit as well as Phe637 and Gly826 in the GluN2B subunit (2,3). Researchers are determining the location of the strongest NMDAR‐ethanol interactions using amino acid substitutions. The Brown Deer High School SMART (Students Modeling A Research Topic) Team has designed a model of NMDAR using 3D printing technology to visualize the NMDAR‐ethanol interactions. If researchers can develop molecules to help minimize the effect of ethanol on the brain, it could reduce the cost of alcoholism to society.

Support or Funding Information

This work was supported by the National Institutes of of Health Science Education Partnership Award (NIH‐SEPA 1R25OD010505‐01) and the National Institutes of Health Clinical and Translational Science Award (NIH‐CTSA UL1RR031973).
Coughing up a Cure for Whooping Cough with Pertussis Toxin
Authors: Tiffany Karen , Arnholt A. , Arnholt S. , Arzumanyan D. , Arzumanyan L. , Butt A. , Houghtaling A. , Ketelhohn L. , Levy J. , Miller N. , Minerva N. , Remington E. , Tiffany B. , Wankowski J. , Barbieri Joseph , Zuverink Madison
Published: November 2023

The CDC reported 32,971 cases of pertussis, or whooping cough, in 2014. This respiratory infection mainly affects unvaccinated infants and toddlers, and symptoms include paroxysmal coughing with whooping, vomiting, and pulmonary complications which can lead to death. Pertussis toxin (PT), produced by the pathogenic bacteria Bordetella pertussis, causes whooping cough by ADP‐ribosylating the Gi protein that inactivates adenylyl cyclase. The inactive ADP‐ribosylated Gi protein is unable to interact with G protein‐coupled receptors, leading to an increased intracellular concentration of cAMP. Increased concentrations of cAMP can interfere with normal cell signaling, disrupting specific cellular functions. PT is a critical virulence factor for B. pertussis that also has potential to create human immunity against pertussis. The Cedarburg SMART Team (Students Modeling A Research Topic) modeled PT using 3‐D printing technology to study structure‐function relationships. PT contains a total of six subunits, four individual subunits (S1, S2, S3, and S5) and a pair of identical subunits (S4). The A domain consists of the S1 subunit, the catalytic portion of the toxin. Mutations in Glu129 and His35 can reduce ADP‐ribosyltransferase activity. Mutations in Arg9, Asp11, and Arg13 also reduce catalytic activity. Trp26 interacts with NAD+, an important cofactor. The B domain, responsible for binding to cell receptors, contains the remaining subunits. Effective vaccines contain chemically‐inactivated PT. However, whole cell pertussis vaccine may cause severe side effects, while acellular pertussis vaccine is not as potent. Non‐toxic derivatives of pertussis toxin may be engineered, improving the potency of the acellular vaccine without harmful side effects.

Support or Funding Information

This work was supported by the National Institutes of of Health Science Education Partnership Award (NIH‐SEPA 1R25OD010505‐01) and the National Institutes of Health Clinical and Translational Science Award (NIH‐CTSA UL1RR031973).
The Relationship between Size, Structure, and Function in Erythrocruorin
Authors: Cabey John , Arroyo Ashley , Sai Aashish Babu , Corpuz Seth , Hoggard Ethan , Huang Kaiyu , Meng Qingqi , Pidara Aneesha , Rehman Iqra , Sabah Clinton
Published: November 2023

Erythrocruorin is a large oxygen‐carrying protein, whose molecular mass is greater than 3.5 million Daltons. This protein is made by earthworms and there are several possible functional reasons why the earthworm erythrocruorin needs to be so large. First, it floats freely in the liquid that flows through their vascular system, and the large size may help qcreduce leakage of hemoglobin. Also, these complex erythrocruorin allow lots of opportunities for interaction between subunits. Finally, huge complexes like this are a way to pack a lot of functional sites into one particle, which can help keep the viscosity of the solution manageable while having a high concentration of sites. The importance of this protein is that since it can be used as a blood substitute in humans‐providing a medical advantage. Erythrocruorin is an effective blood substitute is because it resists oxidation and is very stable.
Evaluation of a Reduced Sampling Schedule for Assessment of Pancreatic Beta‐Cell Function with the Intravenous Glucose Tolerance Test During a Dietary Intervention
Authors: Palacios Orsolya M , Nieman Kristin M , Cook Chad M , Maki Kevin C
Published: November 2023

Pancreatic beta‐cell function is a key determinant of glucose tolerance. The intravenous glucose tolerance test (IVGTT) can be used to assess both the insulin sensitivity index (Si) and the acute insulin response to glucose (AIRg). The disposition index (DI = Si × AIRg) reflects the adequacy of pancreatic beta‐cell response for the level of insulin sensitivity. The aim of this analysis was to compare the AIRg and DI values calculated using a reduced early test sampling schedule (t = 0, 5, 10 min), to those from the full sampling schedule (t = 0, 3, 5, 7, and 10 min). Participants included men (n = 8) and women (n = 16) with mean (SD) age 48.9 (9.9) y and body mass index 30.8 (4.9) kg/m2. Subjects consumed 0, 15, and 30 g/d high‐amylose resistant starch (RS) for 4 weeks each, in a double‐blind, randomized, crossover trial, and underwent a 180‐min IVGTT at the end of each treatment period. The Minmod Millenium program was used to derive Si. AIRg was calculated as the mean increment in plasma insulin during the first 10 min of the test for the full and reduced sampling schedules. The median (interquartile range) full schedule AIRg values during the control, 15, and 30 g/d RS conditions were 43 (29, 103), 47 (23, 67) and 44 (30, 94) mU/L, respectively. Mean (95% confidence interval) percentages of the full schedule values for the reduced schedule values were 82 (79–86), 87 (76–98) and 83 (80–85) for the control, 15 and 30 g/d RS conditions, respectively, all p < 0.05 vs. the full schedule. Correlations between the AIRg and DI values from the two schedules during each RS condition were high (Pearson r values all ≥0.98, p < 0.001). Thus, although AIRg and DI values cannot be directly compared between sampling schedules, the reduced sampling schedule accurately reflects relative AIRg and DI values, and may thus be a time and cost‐effective option when conducting the IVGTT in clinical and epidemiological studies.

Support or Funding Information

Ingredion Inc., Westchester, IL
Consumption of Bifidobacterium animalis subsp. lactis BB‐12 reduces TNF‐α secretion from LPS‐stimulated PBMCs, but does not alter serum TNF‐α levels
Authors: VanEvery Hannah Leigh , Meng Huicui , Flemming Jennifer A , Lee Yujin , Ba Zhaoyong , Furumoto Emily J , Roberts Robert F , Kris‐Etherton Penny M , Rogers Connie J
Published: November 2023

Increasing evidence suggests that probiotic bacteria can modulate inflammatory responses. However the immune response varies by species and strain of organism, as well as delivery matrix of the probiotic. Furthermore the tissue compartment and assay method by which inflammatory mediators are quantified in clinical trials is a debated issue. The goal of the current study was to evaluate the effect of one probiotic species, Bifidobacterium animalis subsp. lactis BB‐12 at a dose of log 10 ± 0.5 CFUs/day on both plasma‐ and in vitro cultured peripheral blood mononuclear cells (PBMC)‐derived inflammatory mediators in a partially blinded, 4‐period crossover, free‐living study. Healthy adults (n=30) aged 18–40 years were recruited, and received 4 treatments in a random order: 1) yogurt smoothie alone; smoothie with organism added 2) before or 3) after fermentation, or 4) organism given in capsule form. At baseline and after each treatment, plasma was collected and PBMCs were isolated and stimulated in vitro with LPS as an inflammatory stimulus. Inflammatory cytokines were quantified in each compartment at baseline and after each treatment. Participants who consumed yogurt smoothies with BB‐12 added post‐ fermentation had a significant reduction in TNF‐α secretion from LPS‐stimulated PBMCs compared to baseline (p=0.039). However, there was no treatment effect observed in plasma TNF‐α levels. Additionally, plasma TNF‐α levels were not correlated with TNF‐α produced by cultured PBMCs in the presence of stimulation. These results demonstrate that quantification of inflammatory mediators in the plasma of healthy adults may not be adequate to detect the effect of probiotic consumption on inflammatory responses. The assessment of a nutritional intervention on in vitro cultured PBMCs in response to LPS stimulation may be a more sensitive assay method to detect probiotic‐induced changes in inflammatory responses. These findings demonstrate that selection of the tissue compartment and the assay method to evaluate inflammatory response is a critical variable in clinical trial design.

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Dairy Research Institute
Estimation of Growth Model Parameters for Mini Pig Breeds
Authors: Jo Hyunwoong , Lee Sang Chul , Kim Dae Cheol , Kim Beob Gyun
Published: November 2023

Pigs are used as models for human because of the similarities in digestive systems. Mini pigs are often used in nutritional studies as well as organ transplantation studies. The relatively small pigs are easier to handle than large pigs. An accurate estimation of body weight of mini pigs is important in animal experimentation. The objective of this work was develop growth models for two mini pig breeds: a M‐Pig® breed originating from Korean Jeju Island native pigs (Cronex Co., Ltd., Hwaseong, Republic of Korea) and Micro‐pig® (Medi Kinetics Co., Ltd., Pyeongteak, Republic of Korea). The M‐Pig® is developed by selecting Jeju Island native pigs with slow growth rate for the purpose of human organ transplantation research. A total 680 body weight data ranging from 0.8 to 63.2 kg for 40 animals were collected from birth to 52 weeks of age. The Micro‐pig® is a crossbreed developed by breeding white gilts selected from cross breeds (Korean native pig, Yucatan mini pig, Vietnamese pot‐bellied pig, and Pygmy pig) with Meishan boars. A total of 36 body weight and age data (mean values of 128 animals) ranging from 0.35 to 33.5 kg and 0 to 96 weeks, respectively, were obtained from 20 published experiments and Medi Kinetics (Pyeongtaek, Republic of Korea). Two different nonlinear growth models with 3 parameters in the equation were employed. Estimated body weight growth models of M‐Pig® as a function of age (week) were: body weight, kg = 64.57 × e−3.21×e^‐0.06t with R2 = 0.976 in Gompertz model and body weight, kg = 57.94 × (1 + 12.33 × e−0.11t)−1 with R2 = 0.969 in Logistic model. Estimated body weight growth models of Micro‐pig as a function of age (week) were: body weight, kg = 31.47 × e−2.10×e^‐0.07t with R2 = 0.793 in Compertz model and body weight, kg = 30.45 × (1 + 4.76 × e−0.10t)−1 with R2 = 0.775 in Logistic model. The age at point of inflection in Gompertz model showed that the maximum growth rate of M‐Pig® and Micro‐pig® was at 18.9 and 11.2 weeks of age, respectively. The body weight at inflection in Gompertz model of M‐Pig® and Micro‐pig® was 23.8 and 11.6 kg, respectively. Taken together, Gompertz models resulted in greater fitness to the growth curve of mini pigs compared with Logistic models. Suggested growth curves for mini pigs may be used for the estimation of body weight gain. Further research is warranted to refine the growth models for mini pigs with a greater amount of data.

Support or Funding Information

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (No. 2016R1A2B2015665).
The Association of Cord Blood 25(OH)D Levels with Neonatal Body Size and Composition: The Healthy Start Study
Authors: Koeppen Hallie J , Kalata Kathryn , Stamatoiu Alexandra V , Ringham Brandy M , Glueck Deborah H , Norris Jill M , Dabelea Dana , Sauder Katherine A
Published: November 2023

Micronutrient deficiencies during pregnancy are associated with an increased risk of chronic disease in offspring. There is some evidence that vitamin D deficiency in pregnancy is associated with increased offspring adiposity, but evidence from human studies is inconclusive. We examined associations of vitamin D (25‐hydroxyvitamin D [25(OH)D]) in cord blood with neonatal size and body composition in 600 mother‐infant dyads from the Healthy Start study, an ongoing, pre‐birth prospective cohort study in Denver, Colorado. Total 25(OH)D (nmol/L) was measured in cord blood serum collected at delivery (39.6±1.1 weeks gestation). Weight‐for‐gestational‐age z‐scores (WGAZ) were calculated from birth weight (g) and birth length (cm) measured at delivery. Neonatal fat mass (g) and fat‐free mass (g) were measured (median 1 day post‐birth) via air displacement plethysmography (PEA POD). We used multivariable linear regression to examine the association of cord blood 25(OH)D with neonatal outcomes, adjusting for maternal age, education, income, race/ethnicity, pre‐pregnant BMI, prenatal smoking, prenatal physical activity, gestational diabetes, gestational weight gain, offspring gestational age at birth, season of birth, and infant sex. We also evaluated effect modification by maternal race/ethnicity and pre‐pregnant BMI. We observed interactions of total 25(OH)D with race/ethnicity for the outcome of offspring birth weight and with maternal pre‐pregnant BMI for the outcomes of offspring birth weight and fat mass (p<0.05 for all interaction terms). The association between birth weight and 25(OH)D was modified by race (p=0.04). The largest effect was observed in Hispanic participants (β= −103.8 grams per 1 nmol/L increase in 25(OH)D, p=0.003) with smaller effects observed among non‐Hispanic white (β= −55.8g, p=0.03), non‐Hispanic black (β= −44.4g, p=0.21), and other (β= −44.4g, p= 0.28) offspring. Associations of total 25(OH)D with birth weight (p=0.02) and fat mass (p=0.04) also differed by maternal pre‐pregnant BMI. Each kg/m2 increase in pre‐pregnant BMI increased the magnitude of the association between 25(OH)D and birth weight by 0.24g per nmol/L 25(OH)D (p=0.02) and between 25(OH)D and fat mass by 0.08g per nmol/L 25(OH)D (p=0.04). Total 25(OH)D was not associated with offspring birth length (p=0.59), fat‐free mass (p=0.61), or WGAZ (p=0.79). In conclusion, we found that total cord blood 25(OH)D is associated with neonatal size and body composition, although the strength and direction of these associations differ according to maternal race/ethnicity and pre‐pregnant BMI. Further research is needed to understand the implications of these associations as infants grow.

Support or Funding Information

Supported by the National Institutes of Health and the American Heart Association
Vitamin D, Calcium Intakes, Physical Activity and Association with Bone Mineral Content Among Adolescents in Malaysia
Authors: Majid Hazreen Abdul Abdul , Aziz Suriawati Abu , Sadat Nabilla Al , Su Tin Tin , Mohd Nahar Azmi , Jalaludin Muhammad Yazid
Published: November 2023

Background

Dietary calcium, vitamin D and physical activity play significant roles in bone development especially throughout children and adolescent growth period. Calcium needs during adolescence increases and the intake of vitamin D may have influence on the BMC.

Learning Outcome

Dietary vitamin D and calcium intake, physical activity (score) and bone mineral content (BMC) among Malaysian adolescents.

Objective

The objective of this study was to determine the dietary intake of vitamin D, calcium and physical activity level with BMC among 13‐year old school‐going Malaysian adolescents.

Method

A cross‐sectional from the Malaysian Health and Adolescents Longitudinal Research Team Cohort Study (MyHeARTs) were conducted among 13‐year old participants of schoolchildren attending Secondary One from central region that includes Kuala Lumpur, Selangor and Perak. The study used a stratified sampling design to select the study participants. Seven‐day diet history were done by trained dietitians and the data were analysed using Nutritionist Pro, using local food database and Singapore Energy and Composition of Food database. BMC of the calcaneus was measured using portable broadband ultrasound bone densitometer (Hologic Sahara, Hologic Inc., USA) as part of the evaluation.

Results

A total of 289 adolescents (65.7% females) were recruited. All subjects had lower intake of vitamin D in comparison with the Recommended Nutrient Intake of Malaysia, 5 μg/day with the mean of 2.51 ± 0.12 μg/day (2.55 ± 0.23 μg/day in male and 2.50 ± 0.14 μg/day in female). Urban adolescents took more food that contained vitamin D than rural counterpart 2.52 ± 0.14 vs 2.46 ± 0.13 μg/day. The average dietary intakes of calcium were 377 ± 12 mg/day (380 ± 14 in urban vs 358 ± 14 mg/day in rural counterpart), respectively, with the majority of subjects failing to meet the Recommended Nutrient Intake (RNI) of Malaysia for dietary calcium of 1000 mg/day. A significant positive relation to BMC was found for dietary vitamin D and calcium intake. All the subjects had a normal Z‐score for the BMC (−2.00 or higher) with a mean of 0.55 ± 0.01 (0.55 ± 0.01 in urban vs 0.54 ± 0.01 mg/day in urban counterpart). Average physical activity score for the participants was categorised as inactive (2.01 ± 0.03).

Conclusions

This study highlights that Malaysian adolescents had inadequate dietary vitamin D and calcium intake and there was no difference between those living in urban or rural. A combination of adequate vitamin D and calcium intake showed positive BMC. It is crucial to promote active lifestyle messages to this age group.

Support or Funding Information

Funding Disclosure: Supported by grants from University of Malaya Research Programme (RP022A,C‐14HTM) and UMSC fund cycle 1‐2016.
Dietary Pattern's Association with Incident Parkinsonism and Progression of Parkinsonian Signs with Aging
Authors: Agarwal Puja , Wang Yamin , Buchman Aron S , Morris Martha C
Published: November 2023

Half of the elderly population by 85 years of age develop mild parkinsonian signs that are progressive with time and are associated with disability and mortality. There are limited studies on how diet may relate to incident Parkinsonism and the progression of this common condition in the elderly. This analysis aims to identify the association of MIND, Mediterranean and DASH dietary patterns with incident Parkinsonism and the progression of parkinsonian signs in older adults.

Methods

A total of 808 participants from the Rush Memory and Aging Project (an ongoing longitudinal cohort study) were assessed annually over 12 years of follow‐up. Parkinsonism was defined as having 2 or more Parkinsonian signs. A trained nurse clinician assessed Parkinsonian signs scores using the modified version of United Parkinson's Disease Rating Scale, including 26‐items that assess four separate signs: Bradykinesia, Gait, Rigidity, and Tremor. MIND, Mediterranean and DASH diet scores were computed using a food frequency questionnaire administered at baseline. Participants with Parkinson's disease at baseline, missing or invalid food frequency questionnaire and less than two Parkinsonian signs assessments were excluded. Incident Parkinsonism was assessed by Cox‐Proportional Hazard ratio. A linear mixed model with random effects was used for Parkinsonian signs scores (square root transformed). All models were adjusted for age, sex, smoking, total energy intake; mixed models were also adjusted for time and its interaction term with covariates.

Results

All three dietary patterns had inverse relation with the risk of Parkinsonism (MIND score (HR=0.87, 95% CI 0.80–0.94), Mediterranean score (HR=0.97, 95% CI 0.94–0.99), and DASH score (HR=0.91, 95% CI 0.83–0.99)). The progression of Parkinsonian signs score was negatively associated with MIND score (β= −0.009; SE=0.0036; p=0.01), Mediterranean score (β= −0.003; SE=0.0013; p=0.02) and DASH score (β= −0.010; SE=0.0046; p=0.03). With further adjustment for depression and BMI, only the MIND diet remained statistically significant for both incident Parkinsonism and progression of Parkinsonian signs. In the final model standardized β coefficients of the estimated diet effects on Parkinsonian signs scores were −2.22 for MIND, −1.75 for Mediterranean and −1.65 for DASH diet.

Discussion

MIND, Mediterranean and DASH dietary patterns are associated with decreased risk of Parkinsonism and may slow the progression of Parkinsonian signs with age.

Support or Funding Information

R01AG054057 and Consolidated Anti‐aging Foundation