bluuebunny/crossref_metadata_2025 huggingface dataset:
Unravelling the chaperonin‐assisted folding of the bacteriophage T4 major capsid protein
Authors: Calmat Stephane , van Heerikhuizen Harm , Schmidt Christoph F. , Peterman Erwin J.G. , van der Vies Saskia M.
Published: June 2021

Folding of bacteriophage T4 major capsid protein, gp23, is essential to the T4 morphogenesis, and is strictly dependent on Escherichia coli chaperonin GroEL in conjunction with the bacteriophage T4‐ encoded co‐chaperonin, gp31. Gp31 is similar in overall structure to the host homologue GroES, which is unable to assist the GroEL‐mediated folding of gp23. Refolding of gp23 in vitro also requires the GroEL‐gp31 chaperonin complex, which encapsulates the major capsid protein in the Anfinsen cage where it acquires its functional structure. We investigate the kinetics of the folding reaction and show that folding is essentially complete after one minute. We reveal novel dynamic information about the molecular mechanism by which the GroEL‐gp31 chaperonin complex facilitates folding. For the first time, the various stages of the folding reaction have been studied in detail. We find that the folding rate of gp23 inside GroEL‐gp31 complex is faster than that of well‐characterized substrates of GroEL‐GroES. Folding of gp23 requires one round of folding i.e. the reaction is very efficient. We conclude that the unique folding abilities of the GroEL‐gp31 chaperonin complex required during bacteriophage T4 infection are conferred by the physiological substrate gp23 in addition to structural properties of the bacteriophage‐encoded co‐chaperonin, gp31.
Dengue virus modulates mitochondrial dynamics during its replication
Authors: Sripada Samatha , Srivastava Nidhi , Dayaraj Cecilia
Published: June 2021

Research over the past few years showed that mitochondrial morphology is a key player in regulation of vital cell physiological processes right from respiration to regulation of apoptosis. Recently the role of mitochondria in virus morphogenesis is gaining importance. In this study, the interaction of dengue virus (DENV) with mitochondria during its replication was investigated, with special emphasis on the association of specific viral proteins with mitochondria. By live cell imaging, it was seen that within an hour of infection, mitochondria showed rapid fission and fusion, indicative of high energy state in the cells. This was followed by recruitment of mitochondria on microtubules to perinuclear region and colocalization with viral factories. As infection progressed, there was circularization and disintegration of mitochondria. In conclusion, our observations suggest a basic dichotomy in the functioning of mitochondria in replication of DENV. Initially, during replication, mitochondria provide energy to ongoing virus replication; while at later stages cells are characterized by mitochondrial dysfunction leading to apoptosis. Of special interest is the implication of viral core protein in induction of apoptosis.

The present study is supported by CSIR (F.NO. 9/698(9)/2004‐EMR‐I) and ICMR, India.
Characterization of Heptameric Anthrax Protective Antigen Containing Phe Clamp Mutations in a Single Subunit
Authors: Janowiak Blythe Elena , Collier R. John
Published: June 2021

Protective Antigen (PA) of anthrax toxin forms a heptameric pore in the endosomal membrane that is responsible for transporting the enzymatic components of the toxin into the cytosol. Previously we showed that effects of mutations in the PA pore could be closely studied using a protocol we devised for isolating heptameric prepores containing a single mutated subunit. The current study uses this new method to ask what precise role the Phe clamp residue (Phe427) plays during translocation. We hypothesized that replacing a single F427 with either a charged residue or a polar residue would result in a large dominant‐negative effect. Thus, we replaced F427 with one of each of the remaining 19 natural amino acids. We prepared monomeric PA containing the test mutation plus an innocuous Cys‐replacement mutation at a second residue in which we biotinylated and used to make and purify heteroheptamers. We found that the largest effect was found when F427 was replaced in one subunit with a charged residue, but we also found reduced cytotoxicity when small hydrophobic residues or polar residues replaced F427 in one subunit. These results suggest that many properties of the phenylalanine are important at this position: being uncharged, having high hydrophobicity and being bulky. This research was funded by NIH (NIAID) (1 F32 AI077280).
The Effects of PBDE‐99 on Lysosomal Membrane Stability in Coelomocytes of the Earthworm Lumbricus terrestris Using the Neutral Red Retention Test
Authors: Sorge Eileen Marie , Clark Matthew
Published: June 2021

Flame retardants, such as polybrominated diphenyl ethers (PBDEs) have recently been found to have toxic side effects due to their bioaccumulative nature. Their widespread use on materials such as electronics, furniture, and plastics has led to contamination of natural environments and exposure to various living organisms, including humans. Lysosomal membrane stability is used as a biomarker of pollutant stressors and is measured using the Neurtral Red Retention test. Neutral Red Retention time measures the length of time it takes red dye to escape from the lysosome into the cytosol, indicating cellular damage. Lumbricus terrestris earthworms were exposed to a medium concentration of PBDE‐99 (50 ng/g dry soil) and a high concentration (250 ng/g dry soil) for one week. After exposure, coelomocytes (gut cells) were extracted using sonication, techniques. Data indicated that treatment with PBDEs resulted in a significant increase in the percentage of cells exhibiting red cytosol per minute. The percent increase in red cells/min. increased from 0.21 to 0.53 from the control to the high group. Decreased lysosomal membrane stability in the high concentration group suggested that PBDEs impaired cellular activity in the earthworm model, potentially making the cells susceptible to degradation by their own hydrolytic enzymes.
The neurosecretory vesicle protein Phogrin is a phosphatidylinositol phosphatase whose activity regulates insulin secretion
Authors: Caromile Leslie Ann , Oganesian Anush , Seifert Ron A , Bowen‐Pope Daniel F
Published: June 2021

Phogrin (IA‐2β) is a 64KD protein localized to the insulin‐containing dense‐core vesicles (DCV) of pancreatic beta cells. Auto‐antibodies against Phogrin are common in pre‐diabetics and are used clinically to diagnose a pre‐diabetic state. Although Phogrin has sequence homology to tyrosine phosphatases, no enzymatic activity has been reported. We report here that Phogrin is able to dephosphorylate specific inositol phospholipids, including PI(3)P and PI(4,5)P2 but not PI(3,4,5)P3. The PIPase activity of Phogrin is low but measurable when a catalytic domain fusion protein is assayed in solution against soluble short‐chain PIPs. Interestingly, when expressed and assayed as a transmembrane protein, Phogrin's in vitro PIPase activity is (100 fold) higher and, when overexpressed in cells, it is able to reduce plasma membrane PI(4,5)P2 levels in a dose‐dependent manner. This activity depends on the catalytic site Cysteine, is inhibited by phosphorylation by PKA, and correlates with inhibition of glucose‐stimulated insulin secretion. Many secretory vesicle‐associated proteins have domains that bind PI(3)P and/or PI(4,5)P2. We propose that Phogrin functions as a phosphatidylinositol phosphatase, that this activity is regulated by phosphorylation by PKA, and plays a role in regulating DCV and plasma membrane PIP content to permit stimulus‐coupled exocytosis of insulin from DCVs.
Novel salivary and genetic biomarkers of risk for NEC or death in premature infants
Authors: Morrow Ardythe Luxion , Jiang Xi , Meinzen‐Derr Jareen , Huang Pengwei , Schibler Kurt , Newburg David , Warner Barbara
Published: June 2021

Objective

Necrotizing enterocolitis (NEC) or death often occur in very low birthweight (VLBW, <1500 g) infants. Intestinal maturation and normal bacterial colonization, which help prevent these outcomes, induce FUT2 ["secretor"] and dampen sialyltransferase gene expression. We tested whether a low secretor, high sialylglycan phenotype predicts risk of NEC or death.

Methods

Salivary secretor and sialylglycans and FUT2 genotype, indirect measures of intestinal phenotype, were analyzed in 385 VLBW infants. Saliva samples collected on day 9 were analyzed by enzyme immunoassay using monoclonal antibodies. Risk cut‐points were identified by Classification and Regression Tree analysis. NEC was defined as Bell's stage >2.

Results

NEC or death occurred in 39 infants. Of infants with low secretor (H‐2), high sialyl (sialyl Lewis a [sLea ], n=104), 23% developed NEC and 18% died; of those with the reciprocal phenotype (high H‐2/low sLea, n=73), no NEC or death occurred; risk was intermediate when H‐2 and sLea were both high or low (p<0.0001; predictive value=0.73). FUT2 genotype was strongly associated with death (13% of non‐secretors [AA], 5% of heterozygotes [AG], 1% of secretor dominant [GG], p=0.001) but not NEC.

Conclusions

Salivary glycans and FUT2 genotype are associated with risk of NEC and death in VLBW infants, and provide powerful biomarkers for clinical research and practice.
DEVELOPMENT OF MULTI‐TURNOVER METALLOPEPTIDES FOR EFFICIENT CLEAVAGE OF HIV‐1 mRNA REV RESPONSE ELEMENT
Authors: Joyner Jeffrey Clark , Cowan James A.
Published: June 2021

Compared to drugs whose mechanism is simple binding to a biological target, multiple‐turnover compounds have potential for much higher efficacy and the ability to harness natural settings. We recently demonstrated catalytic targeting and transformation of a truncated version of the structured Rev response element (RRE) of HIV‐1 mRNA by a class of Arg‐rich peptides which form extremely stable square planar complexes with Cu(II) and Ni(II) cations via an amino‐terminal Cu and Ni (ATCUN) binding motif and bind the RRE with nanomolar affinity via a Rev‐like recognition sequence. If the RRE is cleaved efficiently in an HIV infected helper T‐cell, HIV proliferation may be suppressed by prevention of translation of the viral Env coding region, required for HIV virion infectivity. Under oxidative conditions, catalytic modification of a truncated RRE target by Cu‐peptide complexes has been demonstrated, presumably via the production of reactive oxygen species, which act in a distance dependent manner with highest effect nearest to the Cu center of ROS generation. In this manner, the redox settings of the cell interior may be harnessed to cause catalytic degradation of a targeted mRNA. The anti‐cancer compound bleomycin is believed to degrade DNA in a similar fashion. The multiple‐turnover metal‐ATCUN‐Rev complexes and their derivatives have potential as therapeutic anti‐HIV compounds.
Assessing student understanding in the molecular life sciences using a concept inventory
Authors: Wright Tony , Hamilton Susan , Rafter Mary , Howitt Susan , Anderson Trevor , Costa Manuel
Published: June 2021

Concept inventories are rigorously validated web‐based instruments designed to evaluate the nature and quality of student understandings of key concepts. Such tests provide students and educators with vital feedback about learning and teaching. This poster describes the process used to develop and validate a concept inventory as a diagnostic tool for introductory university courses in the molecular life sciences and is being trialled in three countries. The project has involved the identification of big conceptual ideas unique to the molecular life sciences and captures, in a comprehensive and future‐oriented way, thinking by experts in the field. These were used to identify key concepts that underpin understanding of these ideas. The concepts include; dynamic equilibrium, macromolecular structure, energy transformations, information coding, catalysis and cellular compartmentalisation. Refined sets of questions have been developed for these concepts and tested for validity and reliability.

Students' correct responses to individual items range from 20 ‐ 80%, highlighting specific areas of conceptual weakness. The results to date indicate that although many students entering molecular life science courses can recognise the concepts from enabling sciences, their ability to apply the concepts in new contexts is highly variable.
Plasminogen: The Clot Buster
Authors: Grienke Daniel , Knapp Jessica , Sella Michelle , Hornick Elissa , Jeletz Kelsey , Loberg Kirsten , Panighetti Sara , Tiffany Karen , Bohnsack Richard , Olson Linda , Dahms Nancy
Published: June 2021

Coagulation and fibrinolysis comprise a complex system involving many proteins designed to form fibrin clots when needed to repair a blood vessel and dissolve these clots when no longer needed. One key fibrolytic protein, plasmin, circulates in the blood as plasminogen, an inactive protein made of several domains. Five domains are kringle domains that help plasmin bind to lysine residues in fibrin, while the catalytic domain exhibits protease activity that fragments the fibrin network. Plasminogen is incorporated within the fibrin network as a clot forms, but the catalytic domain is only able to break down fibrin when plasminogen has been activated and converted to plasmin. One substance known to convert plasminogen into plasmin, tissue plasminogen activator (tPA), is a serine protease that cleaves the peptide bond between Arg561 and Val562 in plasminogen. The endothelial cells of damaged blood vessels slowly release tPA that activates the plasminogen embedded within a fibrin clot. Because increased plasmin levels help dissolve clots, tPA is given clinically to treat conditions caused by blood clots, like heart attack and stroke. To further understand the structural implications of activation, a 3D physical model of plasminogen has been designed and built by the Cedarburg High School SMART (Students Modeling a Research Topic) Team using 3D printing technology. Supported by a grant from NIH‐NCRR‐SEPA.
Effect of Ketogenic Diet on Survival of Magnocellular Neurons Following Infundibular Nerve Crush Injury
Authors: Leiphon Laura , King Todd , Veech Richard , Watt John
Published: June 2021

Our experimental objectives were to first determine the effect of two ketogenic diet (KD) formulations on neuroendocrine function and determine the extent to which these ketogenic diets increase plasma ketone levels in rats. Second, we sought to determine the effect of KD and its specific metabolites on alleviation of traumatic brain injury (TBI) ‐induced diabetes insipidus (DI) and the survival of compromised supraoptic magnocellular neurons that are selectively vulnerable to TBI insult.

Methods utilized included immunocytochemical and metabolic analysis, stereometric assessment of neuronal survival, analysis of plasma ß‐hydroxybutyrate and acetoacetate levels, and infundibular nerve crush.

Our results demonstrate that after a 4 week administration of 18% protein KD, plasma ketone levels of β‐hydroxybutyrate and acetoacetate were significantly increased over control diet values in the absence of caloric restriction. Plasma ketone body levels were also significantly increased with 8% protein KD although there was marked caloric restriction associated with this formulation. Interestingly, the 18% protein KD had substantially higher ketone body levels than the 8% protein KD. However, despite increased ketone body levels, no improvement in survival of magnocellular supraoptic neurons was observed following infundibular nerve crush. Nor was there any dietary effect on alleviation of injury induced DI.
PAF receptor activation in hippocampal plasticity induced by SE
Authors: Hutcherson Lori Elizabeth , Musto Alberto Eugenio , Bazan Nicolas Guillermo
Published: June 2021

One of the defining characteristics of epilepsy is abberant mossy fiber sprouting (MFS) in the dentate gyrus. These axonal projections within the granule cell layer of the dentate gyrus create a hyperexcitable network and lead to spontaneous seizures. Growth‐associated protein 43 (GAP‐43), a presynaptic marker that is expressed during development and axonal growth, has been preveiously implicated in the abnormal MFS found in epileptic models. The purpose of our study is to investigate the role of platelet activating factor (PAF) receptor‐mediated signaling in abnormal MFS by evaulating GAP‐43 immunoreactivity. PAF is a mediator of inflammation, a potent gene expression inducer and a modulator of long term potentiation (LTP), synaptic plasticity, and memory formation. PAF abnormally accumulates in the brain during seizures, or injuries, contributing to neuroinflammation and neurotoxicity. Using the novel PAF receptor antagonist, LAU‐0901, in the rat pilocarpine model of epilepsy, we found a positive correlation between a decrease in spontenous seizure activity and an increase GAP‐43 immunoreacitivy in our treated group, suggesting that PAF receptor activation is involved in the abberant plasticity associated with epilepsy.

Source of support: This work was supported by NIH/NCRR grant P20 RR016816.
A Potential Protective Role of RGS2 in Oxidative‐Stress Mediated Anxious Behavior in Rats
Authors: Salim Samina , Taneja Manish , Chugh Gaurav , Saha Kaustuv , Sarraj Nada , Vollert Craig , Tejada‐Simon MariVi , Eikenburg Douglas C , Hovatta Iiris
Published: June 2021

A single nucleotide polymorphism in Regulator of G‐Protein Signaling (RGS)‐2 gene in human and its knockout in mice have been linked to panic disorder and anxiety, respectively. Interestingly, oxidative stress is proposed to be involved in anxiety. The present study is the first to provide a link between RGS2, oxidative stress and anxiety. Our neuronal cell culture studies demonstrate that 1h H2O2 treatment increases while 4h H2O2 treatment decreases RGS2 and antioxidant proteins. RGS2 antisense treatment prevented the increase in antioxidant protein expression suggesting the involvement of RGS2 in this process. Oxidant producing pharmacological agent BSO treatment for 4 and 7 d in rats, increased the levels of oxidative stress markers, protein nitrotyrosine and MDA, in the brain regions implicated in the anxiety response. This was accompanied by a decrease in RGS2 and antioxidant proteins especially, glutathione reducatse 1 and Glyoxalase 1, the proteins previously implicated in the development of anxiety. Two day BSO treatment increased RGS2 and antioxidant protein expression. Furthermore, behavior tests revealed that 4 and 7 d BSO treated rats were more anxious than control or antioxidant, tempol treated rats while 2d treatment did not result in increased anxiety. Our studies suggest an involvement of oxidative stress in anxiety and a novel role of RGS2 in antioxidant homeostasis. [Funding: GEAR, UH].
Effect of Standardized Lycium barbarum (Goji) Juice, GoChi® Intake on Resting Metabolic Rate and Waist Circumference: Randomized, Placebo‐controlled, Double‐blind Clinical Studies
Authors: Amagase Harunobu , Nance Dwight M
Published: June 2021

We examined the effects of consuming Lycium barbarum fruit juice (GoChi®, 120 ml/day) on morphometric parameters and resting metabolic rate (RMR) in healthy human adults in 2 separate randomized, double‐blind, placebo‐controlled clinical studies. GoChi juice is standardized for its main active constituents, Lycium barbarum polysaccharides, based on the amount of fresh berries traditionally used.

Study‐1

Under free‐living conditions, subjects (average age=33.6, body mass index (BMI) =29.3kg/m2) took either GoChi or placebo samples in a 30‐day intervention trial. Waist circumference in the placebo group (N=19) increased significantly by 2.2 cm on Day 30 at post‐intervention compared to pre‐intervention. GoChi group (N=20) maintained pre‐intervention measurements. Changes in waist circumference per caloric intake in GoChi group tended to be lower than placebo group. Body weight and other parameters were not significantly different between groups or pre‐ and post‐intervention.

Study‐2

The effect of a single bout of consuming either GoChi or placebo control samples on RMR was investigated by hand‐held indirect calorimeter (average age=34.5, BMI=28.2 kg/m2). GoChi intake (120 ml) (n=6) significantly increased RMR 53% more than placebo (n=10) at 2 hours after consumption.

These results suggest that GoChi may have metabolic effects which are reflected in waist circumference.
Genetic Disruption of Secreted Renin with Preservation of Intracellular Renin Causes Cardiovascular Dysregulation and Interfered Metabolism
Authors: Xu Di , Borges Giulianna , Yang Baoli , Sigmund Curt D.
Published: June 2021

Secreted renin (sRen) is translated into preprorenin, whereas intracellular renin (icRen) initiates transcription from an alternative promoter, uses a new 1st exon (exon‐1b) and translates from an alternative highly conserved ATG. Since icRen lacks the signal peptide it remains intracellular. We performed gene targeting to ablate sRen while preserving icRen. +/‐ mice carrying the null allele were intercrossed but only two ‐/‐ mice survived (+/+ 34; +/‐ 69; ‐/‐ 2). Daily saline injections of newborns allowed us to rescue additional ‐/‐ survivors that exhibited renal atrophy, lower blood pressure and heart rate, a blunted baroreflex, less brown adipose, and lower body weight. Timed breedings between +/‐ mice were performed to obtain ‐/‐ tissues at 18.5gd. +/+, +/‐ and ‐/‐ fetuses were obtained in expected numbers (7:20:9). Expression of sRen and icRen mRNAs in brain and kidney were detected in +/+ and +/‐ fetuses. ‐/‐ fetuses had no sRen mRNA in kidney and brain, but retained expression of icRen mRNA. We conclude: 1) deleting sRen does not affect icRen mRNA, 2) preservation of icRen is insufficient to rescue lethality caused by loss of sRen, and 3) adult sRen ‐/‐ exhibit cardiovascular and metabolic dysregulation. We are breeding the floxed allele to Nestin/GFAP‐CRE mice to generate brain‐specific KOs of sRen while preserving icREN to examine the physiological relevance of intracellular renin in the brain.
Generation and characterization of a novel, cardiac‐specific, inducible RGS2 transgenic mouse model to modulate Gq‐mediated cardiac hypertrophy
Authors: Park Cindy , Zhu Ming , Chaves Leonard , Zhang Peng , King Michelle , Mende Ulrike
Published: June 2021

Receptor signaling via heterotrimeric Gq proteins leads to hypertrophy, which is a risk factor for heart failure. Among Regulators of G‐protein Signaling (RGS), only RGS2 selectively inhibits Gq signaling and hypertrophy in cardiomyocytes in vitro. Our objective was to generate a mouse model that is suitable to test the hypothesis that RGS2 can modulate hypertrophy development and reversal in vivo. Using a doxycycline (Dox)‐responsive α myosin heavy chain promoter, we generated transgenic mice with cardiac‐specific, conditional RGS2 expression (TG) and established a Dox regimen that prevents RGS2 expression during development and ensures re‐expression by adulthood. Transgenic RGS2 was restricted to the myocardium, localized to both the particulate and cytosolic fraction, and dose‐dependently regulated by Dox in two independent lines. RGS2 expression did not alter heart weight, morphology and function under baseline conditions. Importantly, Gq‐mediated phospholipase Cβ activation was diminished in RGS2‐expressing ventricles. Our data indicate proper expression, regulation and function of transgenic RGS2 with no compensatory changes in other RGS proteins. Thus, this novel transgenic mouse model can now be used to determine the effect of RGS2 on Gq‐mediated pressure overload hypertrophy in vivo, using echocardiography and hemodynamic measurements.

Supported by NIH grant HL80127.
Dietary supplement use among college students
Authors: Carroll Hannah Beth , Spence Marsha Lynn , Paulus Trena M.
Published: June 2021

Dietary supplement use is common among adults. However, little is known about the supplement intake among college students. The purpose of this project was to describe dietary supplement use among college students enrolled in an introductory nutrition course. Of 355 students enrolled in the course at a large southeastern university, 310 students consented to participate in the survey. 307 surveys were complete and used for analysis. Questions pertaining to supplement use, college major, gender, age, BMI, and exercise habits were asked. Preliminary data showed that the sample consisted of 74% females and 26% males. Analysis of exercise habits showed that 59% reported participation in exercise regularly or frequently, 32% reported participation in exercise occasionally, and 8% reported that they rarely/never exercised. About a third of the students (32%) reported taking a multi‐vitamin/multi‐mineral supplement daily. About 30% reported taking some type of herbal supplement. Garlic, cranberry, ginseng, and guarana were most widely used. With nearly 1/3 of college students in this study reporting that they consume dietary supplements on a regular basis, targeted nutrition education regarding the safety and efficacy of supplement use may be an important addition to college‐level introductory nutrition course curricula.
Relationship of uric acid to body composition, heart disease risk factors and energy expenditure
Authors: Kerksick Chad , Beavers K , Chandran R , Jitomir J , Serra M , Shelmadine B , Wilborn C , Wismann J , Nassar E , Dove J , Galbreath M , La Bounty P , Campbell B , Ferreira M , Cooke M , Iosia M , Rasmussen C , Wilson R , Kreider R
Published: June 2021

848 sedentary, obese women (47±11 yrs, 92±18 kg, 163±7 cm, 45±4% body fat) were medically screened prior to participation in an exercise and weight loss program. Medical data and fasting blood samples were collected and a correlation analysis was used to examine relationships between baseline uric acid, body composition and associated risk factors for heart disease. Baseline uric acid levels were significantly correlated to several measures of body composition: body mass (r=0.154), waist circumference (r=0.142), hip circumference (r=0.145), DEXA fat mass (r=0.155), DEXA fat‐free mass (r=0.122) and DEXA % body fat (r=0.106). Higher uric acid levels were also positively correlated with higher triglyceride levels (r=0.091), total cholesterol: HDL ratio (r=0.097) and lower HDL cholesterol (r= ‐0.097). Uric acid levels were positively correlated with baseline measures of resting energy expenditure (r=0.130). Uric acid levels were not significantly correlated with age (r= ‐0.012), resting heart rate (r= ‐0.021), systolic blood pressure (r=0.032), diastolic blood pressure (r=0.055), total cholesterol (r=0.023), LDL cholesterol (r=0.054) and blood glucose levels (r= ‐0.041). Although correlations are modest, data from this sample (n=848) suggest that maintenance of uric acid may be an important consideration in managing obesity and metabolic syndrome.
Food security status, gardening behaviors, and produce intake of low income women with children in Appalachian Ohio
Authors: Jackson Laura , Holben David H , Zurmehly Ashley , Holcomb John P
Published: June 2021

Gardening has been associated with food security among households (HH) with preschool children. This cross‐sectional survey assessed the food security status (FSS), gardening behaviors, and produce intake/behaviors of adult females with school age children in Appalachian Ohio. Participants (n=99) were 34.7±7.2 years and living in HH with 3.8±1.2 members with the majority being white (83.7%) and married (45.9%). HH were classified as fully food secure (n=36, 36.4%), marginally food secure (n=14, 14.1%), low food security (n=24, 24.2%), or very low food security (n=25, 25.3%). 37 (38.1%) participants gardened vegetables, while 23 (24.0%) gardened fruit. FSS did not differ between those who gardened vegetables versus those who did not (p=.482) or between those who gardened fruit versus those who did not (p=.812). FSS was not associated with readiness to garden vegetables (p=.797) or fruit (p=.365). Food insecurity was associated with decreased readiness to eat vegetables (tau b=‐.172, p=.044) and decreased daily intake of vegetables (tau b=‐.224, p=.010). These data support that food insecurity is associated with both decreased vegetable intake and readiness to eat vegetables but not with gardening behaviors. This study supports the need for nutrition and dietetics professionals to promote produce intake and gardening through education programs targeted at improving FSS. Funding: OH Diet Assoc; Am Cancer Soc
The addition of polydextrose and galactooligossacharide to formula does not affect barrier function or bacterial translocation in neonatal piglets
Authors: Monaco Marcia H , Kashtanov Dmitri O. , Rai Deshanie , Walker D. Carey , Jouni Zeina E. , Miller Michael J. , Donovan Sharon M.
Published: June 2021

The microbiota of breastfed and formula‐fed infants differ, largely due to the prebiotic activity of oligosaccharides in human milk. While there is interest in supplementing infant formulas with prebiotics, some studies suggest that prebiotics may increase bacterial translocation (BT). Thus, the objective of this study was to determine the impact of polydextrose (PDX) and galactooligossacharide (GOS), on intestinal BT in neonatal piglets. Colostrum‐deprived piglets (n=36) were randomized to receive total enteral nutrition as formula (TEN); TEN+GOS (4 g/L), TEN+PDX (4 g/L) or TEN+GOS+PDX (2 g/L each) for 7 days. An additional group of piglets (n=8) was placed on total parenteral nutrition (TPN) as a positive control. Blood, spleen, liver and mesenteric lymph nodes were cultured for aerobic (MacConkey and Trypic Soy + 5% sheep blood) and anaerobic (Wilkins‐Chalgren, Bacteroides Bile Esculin and Lactobacillus Agars) bacteria. Bacterial culturing demonstrated significant BT of Enterobacteriaceae, fastidious anaerobes and fastidious aerobes in TPN piglets. In contrast, BT was insignificant in TEN regardless of the presence of prebiotics. Ileal and colonic villus height and width were not affected by prebiotic supplementation (p<0.05), but were affected by TPN. Our study shows that the addition of PDX and GOS to infant formula does not adversely impact BT. (Supported by Mead Johnson Nutritionals)
Stress and religiosity: Modulators for cardiovascular disease risk
Authors: Berk Lee , Bornheimer Joseph , Nelson Bruce , Shaw Sally Fontamillas , Prowse Michelle , King Dana E
Published: June 2021

Background

Clinical evaluation for cardiovascular disease (CVD) typically includes assessment for eight primary symptoms (e.g. chest pain) and hs‐C‐reactive protein (CRP) testing. This study assessed the relationship between stress, religiosity, and factors indicating high CVD risk: at least two reported symptoms and CRP levels >3 mg/L.

Methods

Participants completed a risk survey that integrated traditional and lifestyle psychoneuroimmunology factors, including perceived stress and religiosity defined as regular church, synagogue or temple attendance. A cohort of 1,023 adults, ages 18 to 89, completed an eight‐primary‐symptoms survey. Serum CRP levels were also analyzed.

Results

Those who commonly felt overstressed were twice as likely to report at least two symptoms indicating CVD risk (p<0.01) and 1.4 times more likely to have CRP levels >3 mg/L (p<0.01). Conversely, those who regularly attended church, synagogue or temple were 49% less likely to report at least two symptoms (p<0.01) and 26% less likely to have CRP levels >3 mg/L (p<0.05). When stress and religiosity were combined, the effect on the number of reported symptoms and CRP remained significant (p<0.05).

Conclusion

There may be clinical relevance suggesting that religiosity reduces the negative effect that stress has on individual health in their reported number of CVD symptoms and on elevated CRP levels. UniHealth Foundation grant.
Evaluating the effectiveness of the Personal Nutrition Planner: an online intervention aimed at women ages 21‐65
Authors: Wallace Sharonda P. , Robertson Trina P. , Mouttapa Michele , Weiss Jie
Published: June 2021

The Personal Nutrition Planner (PNP) is an interactive computer based nutrition intervention that provides nutrition recommendations based on age, gender, activity level, ethnicity, risk factors for current disease and current medical conditions. The purpose of this research project was to determine PNP's effectiveness in helping women make nutritious dietary choices.

Participants

female staff members at 2 California State Universities recruited through email advertisement. Study design: pre‐test assessment, the PNP, 4‐weekly emails, one week post‐test, and a post test follow up after 2‐months. A total of 307 women were randomly assigned to control and treatment groups. Areas assessed: psychosocial variables related to diet (stages of change, knowledge items, self efficacy for eating a healthy diet, propensity towards eating a healthy diet); and dietary intake. Posttest minus pretest scores were used to determine changed over time. Many of the scores trended towards significance; yet, no statistically significance outcomes were identified. However, Repeated Measures ANOVA analyses revealed that relative to the control group, the intervention group decreased their intake of sugared beverages and extra foods, increased intake of dairy servings, and lost significantly more weight across the study period. The PNP may have increased obese women's motivation to seek additional assistance regarding food choices relative to other participants in the study. The on‐line Personal Nutrition Planner appears to be a valuable tool for helping women make better dietary choices overall.

This research was supported by the Dairy Council of California
Procyanidin‐ and polysaccharide‐induced γδ T cell priming
Authors: Holderness Jeff S , Hedges Jodi F , Daughenbaugh Katie , Kimmel Emily , Freedman Brett , Robison Amy , Lubick Kirk , Jutila Mark A
Published: June 2021

We recently characterized the capacity of procyanidins and polysaccharides found in common dietary supplements to stimulate innate immunity. Our studies focused on γδ T cells, innate lymphocytes found in mucosal epithelium and well positioned to encounter and respond to agonists found in dietary supplements. Higher order oligomeric procyanidins, such as those found in unripe apple peel (APP) and polysaccharides from plant bark (Yamoa(tm)) primed γδ T cells. Priming of γδ T cells results in rapid increases in innate cytokine expression and enhanced responses to secondary stimuli. Intraperitoneal injection of either procyanidin or polysaccharide agonists induced rapid peritonitis in mice. Feeding Yamoa(tm) extract to calves altered peripheral blood leukocyte responses. Although both procyanidins and polysaccharides primed γδ T cells, they acted via different mechanisms. Whereas procyanidins directly primed γδ T cells, polysaccharides required accessory cells such as monocytes for optimal priming. Procyanidins induced GM‐CSF and IL‐8 production and mRNA stabilization. Our ongoing studies suggest that the responses of γδ T cells may be responsible, to some degree, for the innate immune benefit provided by these dietary supplements. These studies were supported by a PO1 grant from NIH‐NCCAM and an NIH‐NIAID Contract.
Improving analytical reasoning skills in the gross anatomy classroom
Authors: Morton David A , Bradshaw William S , Alder Stephen C , Foreman K Bo , Bell John D
Published: June 2021

Experienced physicians indicate the need to improve analytical reasoning skills as opposed to memorizing volumes of factual minutia. To address this need at the University of Utah School of Medicine we sought to improve the gross anatomy course by implementing active learning exercises in the classroom and increasing the rigor of our exams. Students were instructed to learn the basic facts outlined for each lecture prior to class by studying the provided objectives. Class sessions included a variety of short lectures with additional group application exercises focusing on clinical problem solving scenarios related to the lecture material. The didactic principle was to engage students in active learning exercises such as Socratic dialogue or small group discussions. Homework and midterm exams challenged students to analyze anatomic information in a clinical setting. Results demonstrated that students enjoyed active learning as opposed to passive learning. In addition, students responded well to clinical problem solving as opposed to regurgitation of information. We hope to statistically demonstrate that active learning exercises in the classroom and increased rigor of assessments improve students' analytical reasoning skills.

[Supported by the Fund for the Improvement of Postsecondary Education (FIPSE) and the Dept. of Neurobiology and Anatomy, University of Utah]
Dietary resveratrol induces protective changes in protein levels of transthyretin and drebrin but does not affect plaque burden in a transgenic mouse model of Alzheimer's Disease
Authors: Varamini Behzad , Sikalidis Angelos , Brenna J. Thomas , Bradford Kathryn
Published: June 2021

Objective

To determine protective effects of resveratrol, a polyphenol with anti‐amyloidigenic effects in vivo, in a mouse model of Alzheimer's Disease (AD).

Background

AD is the most common form of dementia, characterized by progressive memory losses with neuritic plaques and hyperphosphorylated tau. Dietary resveratrol decreases amyloid production in vitro but has not been tested in a transgenic mouse model of AD.

Procedure

Fourteen month old wild type and AD transgenic mice were fed either control diet or resveratrol diet in a 2x2 design for 15 weeks to sacrifice. Hippocampal plaque was quantified by immunohistochemistry using MetaMorph 7.1 software. Protein levels were measured using western blot.

Results

Resveratrol diet did not decrease plaque burden in AD mice. Resveratrol‐fed mice demonstrated 2‐fold increases in drebrin, losses of which are associated with clinical AD, and 3.3‐fold increases in transthyretin, a beta‐amyloid scavenger.

Conclusion

Dietary resveratrol did not inhibit plaque formation in this transgenic model of AD, but increased the levels of neuroprotective proteins drebrin, transthyretin, and glycogen synthase kinase 3‐beta‐phospho. Funding: NIH DK07158.
Sodium ferulate modified gene expression profile of oxidized low‐density lipoproteins‐treated endothelial cells
Authors: Zhang Dongxian , Li Yang , Bi Zhuoyue , Wang Chunhong , Ouyang Jingping , Wang Baohua , Bi Yongyi
Published: June 2021

Oxidized low density lipoprotein (ox‐LDL) is shown to activate endothelial cells to induce inflammatory gene expressions. Sodium ferulate, an effective component extracted from Chinese medicines, is known to have a variety of beneficial pharmacological activities. In this study, we investigate the effects of sodium ferulate on gene expression profile of ox‐LDL‐treated endothelial cells. Cultured human umbilical vein endothelial cells were serum starved for 24 hours, then treated with ox‐LDL (50µg/ml) in the absence or presence of sodium ferulate (5ìM). Sodium ferulate's effects o on gene expression profile and ox‐LDL induced cell death were assessed by using human oligonucleotide microarray gene expression chips and cell viability assay, respectively. Sodium ferulate significantly protected cells against the death induced by ox‐LDL. Meanwhile, a total of 32 genes, including members from families of chemokine, inflammatory factor, and growth factor, up‐regulated by ox‐LDL were significantly down‐regulated to the control level by sodium ferulate administration. These data provided an overview of the gene expression profile of endothelial cells in response to ox‐LDL and sodium ferulate, and demonstrated that sodium ferulate could regulate the expressions of inflammation‐related genes in endothelial cells to ameliorate endothelium function related with atherosclerosis that merit further study.
Acute hypoglycemia causes extended anhedonia in mice
Authors: Park Min Jung , O'Connor Jason C. , Lavin Desiree N. , Dantzer Robert , Kelly Keith W. , Freund Gregory G.
Published: June 2021

In humans, hypoglycemia is associated with a variety of mood changes including a reduction in hedonic tone and energetic arousal and an increase in tense arousal. While these clinical symptoms are well characterized, their underlying mechanism is not. C57BL/6J mice were administered insulin that generated a blood glucose nadir of 56 ± 6 mg/dl 0.75 h after injection. At 4 h post insulin administration blood glucose had returned to normal (151 ± 19 mg/dl). Saccharin preference testing 24 h post hypoglycemia showed that insulin receiving mice had saccharin aversion (30 % vs 81 % of total fluid consumption) that took 72 h to resolve. In addition, insulin administered mice (24 h post hypoglycemia) had decreased mobility in the forced swim test and the tail suspension test that took 48 h to rectify. Importantly, activity and burrowing were not impacted 24 h post hypoglycemia. Taken together these data indicate that anhedonia and depressive‐like behavior are induced by hypoglycemia and resolve much more slowly than hypoglycemia, itself, or behaviors associated with locomotion and tunnel maintenance.

This research was supported by grants from National Institutes of Health (DK64862 and NS58525 to G.G.F.) and University of Illinois Agricultural Experiment Station (to G.G.F.).
Oxidative stress in the sympathetic premotor neurons contributes to sympathetic activation in renovascular
Authors: Oliveira‐Sales Elizabeth Barbosa , Nishi Erika Emy , Carillo Bruno Arruda , Boim Mirian Aparecida , Dolnikoff Miriam Sterman , Bergamaschi Cássia Toledo , Campos Ruy R
Published: June 2021

Based on previous data, we hypothesized that an increase of Ang II ‐ via the AT‐1 receptor ‐ in the rostral ventrolateral medulla (RVLM) and the paraventricular nucleus of the hypothalamus (PVN) could activate NAD(P)H oxidase that will produce superoxides resulting in increased sympathetic activity and hypertension. The mRNA expression of AT‐1 receptors, NAD(P)H oxidase subunits (p47phox and gp91phox) and CuZnSOD were analyzed in the RVLM and PVN of male Wistar rats (Goldblatt hypertension model ‐ 2K‐1C). Additionally, we administered Tempol‐1 and 5 nmol into the RVLM, PVN or systemically. The mean arterial pressure (MAP) and renal sympathetic nerve activity (RSNA) were analyzed. The AT‐1 mRNA expression and NAD(P)H oxidase subunits was greater in the RVLM and PVN in 2K‐1C compared to the control group. Furthermore, the CuZnSOD expression was similar in both groups. Tempol‐1nmol into the RVLM reduced MAP (15 ± 1%) and RSNA (11± 2%) only in 2K‐1C rats. Tempol (5 nmol) in the same region decreased the MAP (12± 4%) and RSNA (20 ±7%) respectively, only in 2K‐1C. In the PVN, Tempol‐5nmol resulted in a significant fall in the MAP (24 ± 1%) and in the RSNA (7.9 ± 2%) only in the 2K‐1C. Acute IV infusion of Tempol decreased MAP and RSNA in the 2K‐1C but not in the control rats. The data suggest that the hypertension and sympathoexcitation in 2K‐1C rats were associated with an increase in oxidative stress within the RVLM, the PVN and systemically. Supported by FAPESP.
Attenuated erythrocytic ATP release is caused by extracellular hemoglobin.
Authors: Cole Russell H , Malavalli Ashok , Vandegriff Kim D
Published: June 2021

Local regulation of microvascular flow is a complex process evolved to match oxygen supply to the metabolic demands of the tissue. The RBC has been proposed as an O2 sensor through a direct link between the desaturation of intracellular hemoglobin (Hb) and ATP release, leading to vasodilation1. This theory suggests a link between alterations of O2 transport parameters in the plasma space and vasoactivity. Based on theoretical calculations of O2 extraction from RBCs in the presence of Hb‐based oxygen carriers (HBOCs), we hypothesized that that the addition of cell‐free Hb to the extracellular space provides a supplementary O2 source that reduces RBC desaturation and ATP release. In this study, the total O2 concentration of RBC/HBOC suspensions was lowered by additions of deoxygenated saline, and then assayed for ATP production via the luciferin‐luciferase bioluminescence assay. When an acellular Hb cross‐linked between α subunits (αα Hb, p50 = 33 mmHg) was added to the red cell suspension, ATP production was significantly less in the absence of HBOC. These results provide a possible mechanism for HBOC‐induced vasoactivity through delayed release of red cell ATP.

1 Ellsworth, ML (2000) Acta Physiol Scand 168: 551‐559.
TRAM‐34 stimulates the proliferation of breast cancer cells via activation of estrogen receptors
Authors: Roy Jeremy W , Cowley Elizabeth A , Blay Jonathan , Linsdell Paul
Published: June 2021

We have investigated the effects of specific K+ channel inhibitors on basal and estrogen (E2)‐stimulated proliferation of breast cancer cells. Using the mammary adenocarcinoma cell line MCF‐7 we assayed cell proliferation by [3H]‐thymidine incorporation in the absence or presence of various K+ channel inhibitors with or without E2. Inhibitors of hEAG and hIK K+ channels inhibited basal, but not E2‐stimulated proliferation of MCF‐7 cells. TRAM‐34, a specific inhibitor of hIK channels increased cell proliferation at intermediate concentrations (3‐10 µM), whereas at higher concentrations (20‐30 µM) TRAM‐34 decreased cell proliferation. The mitogenic effect of TRAM‐34 was blocked by the estrogen receptor antagonist ICI182,780. TRAM‐34 also (i) increased progesterone receptor mRNA expression, (ii) decreased estrogen receptor‐α mRNA expression, and (iii) reduced the binding of radiolabelled E2 to MCF‐7 estrogen receptor protein, in each case mimicking the effects of E2. Our results demonstrate that K+ channels hEAG and hIK play a role in basal, but not E2‐stimulated MCF‐7 cell proliferation. TRAM‐34, as well as inhibiting hIK, interacts directly with the estrogen receptor and mimics the effects of E2 on MCF‐7 cell proliferation and gene expression. Our finding that TRAM‐34 may be able to activate the estrogen receptor suggests a novel action of this supposedly specific K+ channel inhibitor. (NSHRF, CBCF)
A Comparative Biomechanical Study of Load Bearing Properties between the Humerus and the Femur
Authors: Khan Hassan S , Hayes Westley , Saha Subrata , Márquez Samuel
Published: June 2021

Living bone is subjected to a series of complex load as external and internal forces act on our skeletal system during locomotion. Differences in mechanical properties are dependent upon whether the bone is weight bearing or not. For example, the human femur exhibits greater mass and resistance to applied force than the humerus presumably due to its role as a weight bearing bone. This quantitative comparative study focuses on load to failure values between the humerus and femur bone tissue. Upper and lower limb segments from cadaveric material were disarticulated and cleaned in order to harvest the bones of interest for this study. Proximal and distal one inch segments were machined and X‐rays were performed to examine for any pathologies. The samples were mechanically tested using the Instron 8874 (Norwood, MA) Servohydraulic Test System to measure the loads to assess compression failure of these segments. Results showed a marked variation in the maximum force between the humerus and the femur. The proximal and distal humeral segments withstood an average compressive load of 18,935 N and 8,440 N respectively. The femoral segments withstood the maximum applied force of 25,000 N. These results confirm weight bearing bones have higher resistance to compressive forces. The results of this study will illuminate the role of load bearing on the mechanical properties of bone tissue.

Grant Funding Source: Dept. of Cell Biology