bluuebunny/crossref_metadata_2025 huggingface dataset:
Avocado Consumption and Risk Factors of Heart Disease: a Systematic Review and Meta‐analysis
Authors: Mahmassani Hiya , Avendano Esther E , Raman Gowri , Johnson Elizabeth J
Published: June 2021

Background

Avocados contain a variety of nutrients and phytochemicals that have individually been related to cardiovascular benefit.

Objective

To summarize the data related to avocado consumption (whole fruit, oil) and risk for cardiovascular diseases (CVD) using systematic review and meta‐analysis.

Design

Medline, Cochrane Central, and CAB databases were searched through October 2016 for all language publications on avocado intake and heart health indices. We included studies of all designs that quantified the amount of avocado intake in adults. CVD risk factors of interest included serum lipids, blood pressure, body weight, blood glucose and insulin levels, inflammatory markers, and all clinical outcomes of CVD. Studies were eligible if participants were healthy or if cardiovascular risk factors such as obesity, dyslipidemia, diabetes, or hypertension were present at baseline. The minimum duration of follow‐up for blood lipid outcomes was set at 3 weeks.

Two investigators independently screened citations and extracted data. Random effect models meta‐analysis was used when at least 3 studies reported the same outcome.

Results

Of the 17 studies (449 subjects) published between 1960 and October 2016 and eligible for our review, 6 studies reported quantitative data for meta‐analyses of avocado intake versus no avocado intake, 3 studies compared avocado plus other monounsaturated fats intake with a control diet, and 8 studies reported qualitative data. The funding sources included industry (5 studies) and government/academia (5 studies) and were not reported in 7 studies. Avocado intake non‐significantly decreased serum total cholesterol (TC) (6 trials, net change −3.97 mg/dL, 95% CI −9.43, 1.50); LDL‐C (6 trials, net change −4.22 mg/dL, 95% CI −11.32, 2.89); and triglycerides (6 trials, net change −20.69 mg/dL, 95% CI −43.32, 1.94). Avocado intake significantly increased HDL‐C (6 trials, net change 3.17 mg/dL, 95% CI 0.21, 6.13); decreased TC:HDL (4 trials, net change −0.45 (95% CI −0.68, −0.22) and LDL:HDL (3 trials, net change −0.19 (−0.38, −0.01). Body weight was unchanged with avocado intake (5 trials, net change 0.19 mg/dL, 95% CI −0.20, 0.57). The differences in outcomes of blood glucose, HOMA‐IR, BMI, systolic and diastolic blood pressure were reported in <3 studies and were not significant between avocado and control groups. None of the eligible studies reported incident clinical outcomes of CVD.

Conclusion

Reviewed studies of avocado intake were unable to show a statistically significant decrease in serum TC, LDL‐C and triglycerides levels. However, avocado intake significantly increased serum HDL‐C levels and consequently decreased TC:HDL ratio and LDL:HDL ratio, indicating a potential association between avocado intake and reduced CVD risk.

Support or Funding Information

This study was supported by the Hass Avocado Board and USDA 1950‐51000‐087
Potential Effects of javamide‐I found in coffee on inflammatory chemokine MCP‐1 in rats fed a high fat diet.
Authors: Park Jae
Published: June 2021

Javamide‐I is a phenolic amide found in coffee with potential health effects. Monocyte chemoattractant protein‐1 (MCP‐1) is a well‐known chemokine and its reduction is considered beneficial in suppressing the pathophysiological progression of cardiovascular, kidney and other diseases, commonly found in the obese. Therefore, in this paper, we investigated javamide‐I as a potential p38 MAP kinase inhibitor, because the kinase is critically involved in the expression of MCP‐1. At the concentration of 20 μM, javamide‐I significantly inhibited p38 MAP kinase alpha, beta, gamma and delta by 15–50% (P < 0.05), particularly p38 MAP kinase alpha (IC50 = 19.2 μM) and beta (IC50 =19.5 μM). We also found that monocytic THP‐1 cells treated with javamide‐I (10, 20 and 40 μM) led to significant inhibition of MCP‐1 production by 17–49% (P < 0.05) in the cells, suggesting that javamide‐I may decrease MCP‐1 expression via inhibiting p38 MAP kinase. Due to the positive in vitro data, the potential effect of javamide‐I on MCP‐1 expression was also investigated in an obesity‐induced rat model. For this study, animals were divided into three groups (n=10): a normal control diet (C), a high fat diet (HF), or a high fat diet supplemented with javamide‐I (2 mg per day) (HFS). After 16 weeks, blood samples were collected from the animals in each group and the plasma levels of MCP‐1 protein were determined. As expected, the average MCP‐1 level of the HFS group was significantly lower than that of the HF group (P < 0.05), although the average MCP‐1 level of the HF group was found to be higher than that of the C group (P < 0.05). In summary, the data suggest that javamide‐I found in coffee may inhibit MCP‐1 expression, possibly via inhibiting p38 MAP kinase and its supplementation could suppress plasma MCP‐1 expression critically involved in several inflammatory diseases.

Support or Funding Information

USDA (1235‐51000‐054‐00D)
Synthetic Cannabinoids: Rapidly Emerging Drugs of Abuse
Authors: Radominska‐Pandya Anna , Moran Jeff , Fantegrossi William , Prather Paul
Published: June 2021

K2, also called “Spice” or “synthetic marijuana,” is a rapidly emerging drug of abuse that possesses psychoactive properties similar to those of Δ9‐tetrahydrocannabinol (Δ9‐THC). K2 use has exploded in many sections of the population including teenagers and first time drug users. Use of K2 can result in extreme agitation, hallucinations, supraventricular tachycardia, syncope, and seizures. The presence of more than 20 different K2‐aminoalkyl indoles (AAIs) have been reported in various K2s, but the two most commonly observed are JWH‐018 and JWH‐073; however, new generations of structurally related compounds are constantly being produced. Our studies demonstrate for the first time that the native K2s undergoes extensive metabolism by cytochrome P450s and UDP glucuronosyltransferases. Due to the activity of these enzymes, a variety of hydroxylated metabolites, have been biosythesized and excreted in human urine primarly as glucuronidated conjugates. These metabolites were identified and characterized using LC‐MS/MS and HPLC‐UV/Vis, and steady state kinetic analyses were also investigated. Consequently, we hypothesize that the severe effects observed for some K2 users could be related to a defect in their metabolism. We have also shown that these K2 products cause psychoactive effects similar to those of Δ9‐THC by activating cannabinoid receptors (CBRs) in the central nervous system. Moreover, CBRs were able to bind several hydroxylated and glucuronidated K2‐AAI metabolites with an affinity similar to that of the parent compound. The new generation of K2s like AB‐Pinaca exhibit higher affinity, potency, and efficacy at CBRs relative to THC. Furthermore, phase one metabolites of AB‐Pinaca retain affinity and full agonist activity at CBRs. Chronic treatment of cells with AB‐Pinaca produces greater desensitization of CBRs (e.g., tolerance) than similar exposure to THC. The higher affinity and activity of AB‐Pinaca at CBRs relative to THC might be expected to produce enhanced psychotropic and adverse effects, contributing to the reported abuse and marked toxicity of this novel SCB. Finally, our in vivo data demonstrates that K2 metabolites retain biological activity in mice. Therefore, we postulate that synthetic cannabinoids and their derivatives could act as ligands for CBRs. The fact that some hydroxylated derivatives and their glucuronides can retain their biological activity makes the study of these compounds essential for understanding their severe toxicity and pharmacokinetics/dynamics.

Support or Funding Information

[DoD‐W81XWH1110795, NIH/NIDA DA039143 to ARP, PLP, WF and JM]
Tissue Inflammatory Responses in an Anesthetized Rat Hemorrhage Model Associated with Endothelial Dysfunction
Authors: Dubick Michael A , Torres Luciana N , Barr Johnny L , Torres Filho Ivo P
Published: June 2021

We have demonstrated endothelial dysfunction such as reduction in glycocalyx thickness and increased permeability in venules in cremaster muscle following hemorrhage and fluid resuscitation with a crystalloid or a colloid in a rat model. The present study further characterizes the model by measuring indices of inflammation and oxidative and nitrosative stress in tissues from these animals. Anesthetized Sprague‐Dawley rats (n=4–5/group) were bled 40% of their blood volume, subjected to a 30 min shock period and resuscitated with limited volumes of normal saline (NS) or 5% albumin (A), or no treatment (Hem). At 1hr after resuscitation, rats were euthanized and liver, kidney, lung and skeletal muscle were collected and assayed for cytokines and select indices of nitrosative and oxidative stress. In liver and kidney, IL‐1β, IL‐6 and TNF‐α levels were 2–3‐fold higher in the NS than the A group and about 2‐fold higher than the Hem group. IL‐10 levels were also significantly higher in the NS group than the other groups. Similar responses were observed in lung from these groups, while in muscle, only IL‐1β and IL‐6 levels were significantly higher. In contrast, no significant differences were observed in all tissues in nitric oxide, total antioxidant or reduced glutathione concentrations nor xanthine oxidase activity among the groups. In this model of hemorrhagic shock, NS induced the highest inflammatory response, while 5% albumin was associated with the least response. Interestingly, these current observations were related to the degree of endothelial dysfunction observed previously. Taken together these data suggest that the endothelial dysfunction observed in hemorrhagic shock and fluid resuscitation is associated with an inflammatory response without a major nitrosative or oxidative stress component.

Support or Funding Information

Funded by the US Army Medical Research Materiel Command
GABA and Glycine are Co‐released into Dorsal Motor Nucleus of the Vagus Neurons following a Perinatal High Fat Diet
Authors: Clyburn Courtney Elizabeth , Browning Kirsteen N.
Published: June 2021

Efferent motoneurons of the adult dorsal motor nucleus of the vagus (DMV) have previously been shown to only receive GABAergic inhibitory synaptic inputs. In contrast, it has been demonstrated that, at early postnatal time points, motoneurons of the DMV receive both GABAergic and glycinergic inhibitory synaptic inputs. The developmental decline in glycinergic input has been determined to occur in the second postnatal week. Our previous work has suggested that rats that are fed a perinatal high fat diet (pHFD) do not undergo this developmental shift and maintain a glycinergic input into adulthood. The purpose of this study was to test the hypothesis that glycine and GABA are co‐released from the same synaptic terminal in pHFD rats.

Whole cell patch clamp recordings were made from DMV neurons in pHFD or control fed Sprague‐Dawley rats 4–6 weeks of age. These rats were fed either a control chow or HFD (14 or 60%kcal from fat, respectively) from embryonic day 13. The effects of the benzodiazepines (BZD) clonazepam (100nM), an allosteric modulator of the a2/3 subunit containing GABAA receptor, and zolpidem (1μM) an allosteric modulator of the a1 subunit containing GABAA receptor, on miniature postsynaptic inhibitory currents (mIPSCs) were examined. Application of BZDs is known to modulate the open probability of ligand‐gated GABAA, but not glycine, ion channels, thus increasing the decay of the slower GABAergic, but not the faster glycinergic mIPSCs. This allows for the differentiation of mixed concurrent glycinergic and GABAergic currents through the identification of two distinct rates of decay.

Application of clonazepam significantly increased the proportion of two‐phased mIPSCs in the DMV of pHFD rats (76/200 vs 14/213 n=6 neurons, p<0.05), but not in controls (10/156 vs 11/164, n=6 neurons, p>0.05). Similarly, application of zolpidem significantly increased the proportion of two‐phased mIPSCs in the DMV of pHFD rats (60/179 vs and 14/153, n=6, p<0.05) but not in controls (27/150 vs and 8/138, n=5, p>0.05).

This study confirms our previous studies suggesting that exposure to a pHFD prevents the developmental decline in glycinergic currents onto the DMV neurons and further suggests that glycine and GABA are co‐released from the same synaptic terminal. Furthermore, significant differences were not apparent between the effects of zolpidem and clonazepam, suggesting that a1, 2 and 3 receptor subunits form functional GABAA receptors in pHFD DMV neurons. These developmental alternations in inhibitory signaling in brainstem motoneurons may result in the dysregulation of gastrointestinal reflexes and gastric motility observed in these rats.

Support or Funding Information

Funded by NIH 078364 and NSF IOS1148978
Phrenic Motor Neuron Survival Caudal to C2 Hemisection
Authors: Allen Latoya Lynn , Santiago Juliet V , Asa Zachary A , Holland Ashley E , Ciesla Marissa , Satriotomo Irawan S , Mitchell Gordon S , Gonzalez‐Rothi Elisa Janine
Published: June 2021

Cervical spinal injury (cSCI) severs descending brainstem projections to spinal respiratory motor neurons, paralyzing respiratory muscles caudal to injury. Spinal hemisection at C2 (C2Hx) has frequently been used as an experimental model of cSCI to study both spontaneous and induced functional recovery of both respiratory and non‐respiratory somatic motor function after cSCI. One assumption implicit in these studies is that C2Hx dennervates motor neurons below the injury, but does not cause significant motor neuron pathology or cell death. However, one recent study reported substantial, bilateral motor neuron death caudal to C2Hx (Satkunendrarajah et al, 2016); in this study, motor neurons were identified by ChAT immunoreactivity in the spinal ventral horn. We have also observed that retrograde labeling with Cholera toxin B fragment (CtB) is diminished in phrenic motor neurons following C2Hx (unpublished data). However, these observations are potentially misleading since: 1) C2Hx may diminish ChAT immunoreactivity without motor neuron cell death; and 2) retrograde transport of CtB may be diminished when administered following an injury, due to the relative inactivity of motor neurons below the injury, giving the false impression that phrenic motor neurons had died. Because phrenic motor neuron cell death following cSCI would have profound implications for studies of spontaneous and/or induced phrenic motor plasticity after C2Hx, we tested the hypothesis that C2Hx has minimal impact on phrenic motor neuron cell counts if phrenic motor neurons are retrogradely labeled with CtB PRIOR to spinal injury. Phrenic motor neurons were retrogradely labeled in intact adult Sprague Dawley rats via intrapleural injections of CtB. Rats were randomly assigned to one of four groups: 1) 2 week C2Hx, 2) 8 week C2Hx, or 3) age matched, spinal intact controls. One week following the CtB injection, rats in the C2Hx groups underwent a left lateral C2Hx injury under isoflurane anesthesia. Rats were then perfused at 2 or 8 weeks post‐injury, and longitudinal sections (40 μm) from C3 to C8 were processed to reveal CtB, ChAT and NeuN. CtB labeled phrenic motor neurons were quantified using a custom MATLAB code. Preliminary analyses suggest minimal loss of phrenic motor neurons at either time post‐C2Hx relative to spinal intact rats. Assessment of ChAT and NeuN immunoreactivity in phrenic motor neurons after C2Hx is ongoing. Although we found little evidence for phrenic motor neuron cell death caudal to C2Hx, we cannot rule out greater cell loss in other motor neuron pools. These findings have important implications for studies concerning motor plasticity following cervical spinal injury.

Support or Funding Information

Supported by: T32HD043730, K12HD055929 (EGR), RO1HL69084 (GSM), and grants from the DoD (GSM), and the McKnight Brain Institute (GSM, MCC)
Characterization of the Receptors Associated with the Differing Responses to the Neuropeptide, AST‐C, by the Cardiac Ganglion of the American Lobster, Homarus americanus
Authors: Walsh Patrick , Pong Sovannarath , Armstrong Matthew , Christie Andrew E , Dickinson Patsy S
Published: June 2021

Central pattern generators (CPGs) are neural networks that produce consistent, rhythmic patterned outputs to reliably activate specific muscles, resulting in repeated and rhythmic movements. The cardiac ganglion (CG) is a CPG in the American lobster, and is responsible for producing a consistent rhythmic heartbeat. Flexibility of rhythmic motor outputs arises from modulation of CPGs, largely by neuropeptides. The lobster heart is modulated by C‐type allatostatins (AST‐C), two of which [pQIRYHQCYFNPISCF, now AST‐C I, and SYWKQCAFNAVSCFamide, now AST‐C II] were previously known; both have been shown to modulate the lobster heart. Using transcriptomes generated from nervous system tissues, we identified a third isoform of AST‐C, GNGDGRLYWRCYFNAVSCF, or AST‐C III, in the lobster. All three AST‐C isoforms consistently lead to decreases in contraction frequency when perfused through an isolated lobster heart; in contrast, all can elicit variable responses in contraction amplitude, with amplitude increasing in some lobsters and decreasing in others. The responses to AST‐C I and AST‐C III are more similar to one another than to AST‐C II in any given lobster.

We are currently testing the hypothesis that differences in AST‐C receptor expression among individuals underlie these varying responses. Mining of a Homarus americanus transcriptome has led to the identification of three full‐length and one partial putative AST‐C receptors. We are currently conducting experiments to determine whether and to what extent the four putative AST‐C receptors bind the three AST‐C isoforms. Understanding these receptor‐peptide relationships will allow us to elucidate the role the receptors play in the observed differential response to AST‐C.

Support or Funding Information

NSF (IOS‐1353023, IOS‐1354567), NIH (8P20GM103423‐12), Cades Foundation, Doherty Foundation gift to Bowdoin College
Linking morphology and motion: Testing multibody simulations against in vivo cranial kinematics in suction feeding fishes using XROMM
Authors: Olsen Aaron , Hernandez Patricia , Camp Ariel , Brainerd Elizabeth
Published: June 2021

The heads of ray‐finned fishes contain over 20 mobile skeletal elements, interconnected by a morphologically and functionally varied set of articulations that allow a high degree of mobility. Many fish use this mobility to expand the buccal cavity during feeding to create suction and draw in prey items. Due to this high degree of mobility, a varied set of articulations, and the diversity of cranial forms among fishes, fish skulls provide an excellent system for relating skeletal and articular morphology to skeletal motion. The motion of fish skulls is traditionally simulated using planar mechanical linkage models, such as the 4‐bar linkage. While linkage models accurately predict the in vivo kinematics for particular subsets of bones within the skull, there are other aspects of cranial motion for which planar 4‐bar linkage models have proven insufficient. Additionally, these models have rarely been tested against three‐dimensional kinematics. We used CT data and the R package linkR to build multibody simulations of cranial motion in three species of ray‐finned fishes: largemouth bass (Micropterus salmoides), channel catfish (Ictalurus punctatus), and common carp (Cyprinus carpio). For each species, we created a series of simulations with varying numbers of parameters and degrees of freedom. We then tested the accuracy of these simulations against three‐dimensional in vivo kinematics, collected using X‐ray Reconstruction of Moving Morphology (XROMM). In evaluating model accuracy, we apply a new approach that measures how model error changes as a function of model complexity. We demonstrate the potential, and limitations, of using morphology to predict in vivo kinematics and ultimately to test how the evolution of organismal form influences the evolution of organismal motion and performance.

Support or Funding Information

This work was funded by NSF DBI‐1612230.
DNA Polymerase η Is an &[Iota]mportant &[Beta]ypass DNA Polymerase in the Repair of DNA Interstrand Cross‐Links
Authors: Zhao Linlin
Published: June 2021

DNA interstrand cross‐links (ICLs) are cytotoxic DNA lesions derived from reactions of DNA with a number of anti‐cancer reagents as well as endogenous bifunctional electrophiles. Deciphering the DNA repair mechanism of ICLs is important for understanding the toxicity of DNA cross‐linking agents and for the development of effective chemotherapies. The repair of ICLs in mammalian cells involves sequential endonucleolytic excision of the lesion from one strand and then the other, and translesion synthesis (TLS) is a critical step for bypassing the unhooked intermediates from the nontemplate strand or for conducting postreplication gap‐filling DNA synthesis. A number of TLS DNA polymerases in humans have diverse bypass specificities and replication fidelities. To illustrate the important role of a certain TLS in ICL repair, we used model ICLs induced by oxidized abasic lesions and those‐derived from O6‐2′‐deoxyguanosine. We investigated the DNA replication‐blocking and mutagenic properties of these ICLs during TLS using primer extension assays and steady‐state kinetic analysis of single nucleotide incorporation prior to, opposite and past the cross‐linked template nucleotide. Liquid chromatography‐mass spectrometry‐based oligonucleotide sequencing was also used to determine the relative yield of the mutagenic bypass products. Our results demonstrated that among the several human TLS DNA polymerases tested, pol η exhibits the relatively high bypass activity; however, a significant portion of bypass products is mutagenic and contains substitution or deletion mutations. The importance of pol η in ICL bypass is demonstrated using whole cell extracts of Xeroderma pigmentosum variant patient cells and those complemented with pol η. Together, this body of work provides in vitro evidence of the important role of DNA polymerase η in ICL repair and underscores the importance of further evaluating of the role of pol η in ICL repair pathways in biological systems.

Support or Funding Information

This work was supported by in part by Central Michigan University start‐up funds, Central Michigan University Faculty Research and Creative Endeavors Award and a grant from U.S. Army Research Office (W911NF‐15‐1‐0140).
Interactions and Dynamics Within CHIP/Hsp70 Complexes
Authors: Mannion Matthew M , Zhang Huaqun , VanPelt Jamie , Cottingim Kelsey , Tsutakawa Susan , Page Richard C
Published: June 2021

Interactions between the E3 ubiquitin ligase CHIP (C‐terminus of Hsp70 interacting protein) and the chaperone Hsp70 represent the physical intersection of two competing protein quality control efforts: refolding, and degradation. Interaction with CHIP allows for the formation of a productive encounter complex that facilitates ubiquitination of Hsp70‐bound substrates, thereby targeting those proteins for proteasomal degradation. Central to the CHIP/Hsp70 interaction are changes in structure and dynamics that occur upon binding. Although recruitment of Hsp70 to CHIP via interaction with the CHIP tetratricopeptide repeat (TPR) domain is well established, alterations in structure and dynamics of CHIP and Hsp70 upon binding are not well understood. Using NMR we describe residue‐level changes in dynamics to the CHIP‐TPR domain that occur upon binding Hsp70. We also utilize small angle X‐ray scattering (SAXS) and electron paramagnetic resonance (EPR) to identify motions with the CHIP/Hsp70 complex. Independent SAXS and EPR experiments both point to a mobile Hsp70 substrate binding domain that is only partially constrained through interactions with the CHIP‐TPR. Building upon static models of the full‐length CHIP/Hsp70 complex, afforded by our recent crystal structure of the CHIP‐TPR/Hsp70‐lid‐tail complex, we have assembled a dynamic model of the CHIP‐Hsp70 complex. This dynamic model sheds light on the function of the CHIP/Hsp70 chaperoned ubiquitination complex and how CHIP harnesses dynamics of Hsp70 to overcome significant distance separations identified in earlier static models of the CHIP/Hsp70 complex.

Support or Funding Information

The authors acknowledge financial support from the US National Science Foundation (Award No. MCB 1552113 to RCP), the American Heart Association (Award No. 16SDG26960000 to RCP), the Burroughs Wellcome Fund (Award No. 1014031 to RCP), and institutional support from Miami University through the Robert H. and Nancy J. Blayney Professorship (to RCP). The Advanced Light Source is supported by the US Department of Energy under contract number DE‐AC03‐76SF00098 at Lawrence Berkeley National Laboratory.
The Mechanism of Inhibition of Botulinum Neurotoxin Type A by Two Quinolinol Compounds
Authors: Minnow Yacoba Vroom Teschemaker , Goldberg Ronald , Tummalapalli Sreedhar R. , Rotella David P , Goodey Nina M
Published: June 2021

The potential of Botulinum neurotoxins (BoNTs) to be used as biological weapons makes the need to identify small molecule inhibitors critical but it is a challenging target due to the large substrate‐enzyme interface. BoNTs are produced by the bacterium Clostridium botulinum in seven antigenically distinct serotypes (A‐G), with serotype A (BoNT/A) being the most potent toxin known to man. Recently, quinolinol‐based compounds have been identified as a promising starting point for effective BoNT/A inhibitors. Understanding the mechanism of inhibition by quinolinol compounds would facilitate inhibitor optimization studies. In this study, a Foster resonance energy transfer (FRET) based substrate of BoNT/A, SNAPtide, was used to study the mechanism of inhibition of BoNT/A endopeptidase activity by two new quinolinol compounds, MSU58 and MSU84, with IC50 values of 3.3 μM and 5.8 μM, respectively. Kinetic analysis and model discrimination analysis revealed both compounds to be competitive inhibitors of BoNT/A endopeptidase activity. Inhibition constants (KI) were found to be 3.2 μM for MSU58 and 6.2 μM for MSU84. The kinetic rate constant for the binding and release of substrate and inhibitor were also determined. These data indicate that these inhibitors bind in the active site of BoNT/A and that binding is mutually exclusive with the binding of the substrate. This study, is the first to report on the competitive inhibition of BoNT/A by a quinolinol group of compounds. These data help define the inhibitor binding pocket and, along with structure activity relationship studies, provide immediate direction for further compound synthesis.

Support or Funding Information

Novartis Graduate Scholarship Award and US Army Medical Research (grant number W81XWH‐14‐C‐0090 S)
Phosphoproteins in Extracellular Vesicles as Candidate Markers for Breast Cancer
Authors: Tao Weiguo Andy , Chen I‐Hsuan , Iliuk Anton
Published: June 2021

The state of protein phosphorylation can be a key determinant of cellular physiology such as early stage cancer, but the development of phosphoproteins in biofluids for disease diagnosis remains elusive. Here we demonstrate, for the first time, a strategy to isolate and identify phosphoproteins in extracellular vesicles (EVs) from human plasma as potential markers to differentiate disease from healthy states. We identified close to 10,000 unique phosphopeptides in EVs isolated from small volumes of plasma samples. Using label‐free quantitative phosphoproteomics, we identified 144 phosphoproteins in plasma EVs that are significantly higher in patients diagnosed with breast cancer as compared to healthy controls. Several novel biomarkers were validated in individual patients using Paralleled Reaction Monitoring for targeted quantitation. This study demonstrates that the development of phosphoproteins in plasma EV as disease biomarkers is highly feasible and may transform cancer screening and monitoring.

Support or Funding Information

This study was supported partially by NIH grants 5R41GM109626, 1R01GM111788, and 1R41CA210772 and by NSF grant 1506752.
Novel role of bHLH proteins in synaptogenesis: Class I bHLH proteins TCF4 and Daughterless restrict synaptic branching and bouton formation via Neurexin repression in postmitotic neurons
Authors: Robinson Erin L. , Waddell Edward A. , D'Rozario Mitchell , Marenda Daniel R.
Published: June 2021

Basic helix‐loop‐helix (bHLH) proteins encompass a highly conserved superfamily of transcription factors that serve as regulators of many developmental processes including neurogenesis. This protein family is divided into six subclasses based on tissue specification, ability to bind DNA at E‐box consensus sequences, and ability to form dimers. Transcription factor 4 (TCF4) is a class I bHLH protein which functions by homo‐ or hetero‐dimerizing with class II/V tissue‐specific bHLH binding partners to control target gene transcription. Haploinsufficiency of TCF4 has been identified as the causative factor in the rare autistic‐like neurodevelopmental disorder, Pitt‐Hopkins Syndrome (PTHS). Similarly, SNPs located in introns of TCF4 have been implemented as a risk factor associated with the psychiatric disorders Schizophrenia and Major Depressive Disorder. Despite the well‐known functions of class I bHLH proteins during development, our laboratory has identified a novel, conserved role for both Tcf4 in mice and its homolog in Drosophila melanogaster, Daughterless (Da), with which they are functioning in postmitotic neurons to restrict synaptic branching and bouton formation via restricting transcription of the presynaptic cell adhesion protein, Neurexin. Synaptic branching is one of the ways that neurons remain plastic post development and aberrant branching of the synapses has been associated with defects in neuroplasticity. This suggests that class I bHLH proteins may have a role in neuroplasticity apart from their distinguished role in neurodevelopment.

Support or Funding Information

NSF Grant: IOS1256114
Whole Grain Gluten‐Free High Protein Buckwheat‐Kale Snacks
Authors: Kahlon Talwinder Singh , Avena‐Bustillos Roberto J , Chiu Mei‐Chen Maggie
Published: June 2021

Four kinds of whole grain gluten‐free high protein Buckwheat‐Kale snacks were evaluated. The snacks were Buckwheat‐Kale (BWK), BWK‐Garlic, BWK‐Onion and BWK‐Ginger. Peanut meal as low value agricultural byproduct was utilized to increase the protein content of these snacks. The levels of dry garlic, onion and ginger were decided by consensus of laboratory personnel. The snacks were low in fat and salt. Snack dough was prepared using water nearly 1:1 as other ingredients. About 20 g of snack dough was placed on preheated Krum Kake Express 839 Baker and cooked for 2 minutes. Sixty nine in‐house volunteers judged Taste/Flavor significantly higher for BWK, BWK‐Garlic and BWK‐Onion snacks than for BWK‐Ginger snacks. Odor/Aroma for BWK‐Garlic snacks was significantly higher than for BWK‐Ginger snacks. Acceptance for BWK and BWK‐Garlic snacks was significantly higher than for BWK‐Ginger snacks. Water activity of all the snacks tested ranged from 0.35–0.38 suggesting that these snacks were crispy and would have good anti‐microbial stability. These snacks were quite light and brittle due to their expansion of 3.6–4.2 times and high (0.72–0.75%) porosity. Acceptance of snacks tested was BWK and BWK‐Garlic 94%, BWK‐Onion 86% and BWK‐Ginger 78%. These snacks contained only 4–5 ingredients and could be made in any house kitchen or commercial production. Acceptance of 78–94% is very desirable. These high protein healthy nutritious snacks offer choice for all consumers, including individuals sensitive to gluten.

Support or Funding Information

USDA‐ARS base funding
Farmers' market use is associated with fruit and vegetable consumption among low‐income populations: A cross‐sectional study from Southern Arizona
Authors: Alaofe Halimatou , Downs Navaugh , Jones Kara , Plano Abigail
Published: June 2021

Background

While farmers' markets are identified as a strategy to increase access to fruits and vegetables (F&V), more information is needed regarding use of farmers' markets in low‐income populations. Thus, this study's purpose was to examine (1) socio‐demographic characteristics of participants; (2) barriers and facilitators to farmers' market shopping and F&V consumption; and (3) associations between farmers' market use with F & V consumption in southern Arizona.

Methods

Cross‐sectional surveys were conducted among 2 groups of Supplemental Nutritional Assistance Program (SNAP) recipients in Pima County, AZ. The farmers' market (FM) customers were interviewed using an intercept survey instrument at 2 farmers' markets while the primary household food shoppers (non‐FM) were randomly identified at Community Food Bank warehouse. All questionnaires assessed socio‐demographic characteristics, food shopping patterns, barriers to and facilitators of farmers' market shopping and F&V consumption. Linear regression analyses were used to examine associations between farmers' market use with F&V consumption, controlling for age, race, education, and gender.

Results

Our findings suggest that FM customers are more likely to be white, female, over 40 years of age and highly educated. Among FM customers, 45% reported shopping at a farmers' market at least weekly while the top 3 main motivators for shopping included variety of products, fresh food and convenient location. Frequently reported barriers to F&V consumption were price, lack of nearby options, and lack of available healthy options. Additionally, 32% and 7.5% of FM customers respectively consumed F&V less than one time per day, respectively. Finally, fruit and vegetable consumption was positively associated with use of farmers' markets.

Conclusions

Farmers' markets may be a viable method to increase produce consumption in low‐income population. However, these programs to be successful should take into account barriers and facilitators to farmers' market shopping and F&V consumption.

Support or Funding Information

USDA FINI Grant Program
First trimester maternal adiposity is associated with infant body fat at age 2 weeks: a longitudinal follow‐up study
Authors: Andres Aline , Ruebel Meghan L , Krukowski Rebecca A , Badger Thomas M , Shankar Kartik , Cleves Mario A
Published: June 2021

Maternal obesity has been shown to be associated with childhood obesity risk in both experimental and epidemiological studies. However, longitudinal studies carefully evaluating other contributors to fetal and infant body fat accretion are lacking. The objective of this study was to evaluate the effect of maternal adiposity on fetal fat mass accretion while controlling for gestational weigh gain, physical activity, basal metabolic rate, insulin sensitivity and depression throughout pregnancy. Pregnant women were enrolled in the Glowing Study before gestation week 10 (95 normal weight BMI and 113 overweight and obese BMI). Adiposity (Air Displacement Plethysmography, ADP), basal metabolic rate and respiratory exchange ratios (indirect calorimetry), physical activity (total activity counts by accelerometry), insulin sensitivity (ISHOMA) and maternal depression (Beck Depression Inventory, BDI) were evaluated during the first trimester of gestation. Body weight was evaluated every 6 weeks throughout gestation and at each visit participants were provided with counseling to assist them with adhering to the Institute of Medicine gestational weight gain guidelines. Infant body fat mass was measured using ADP at age 2 weeks. Multiple regression analyses demonstrate that maternal adiposity (% body fat) is significantly associated with infant body fat (%) at age 2 weeks (β=0.15, P=0.0004) when controlling for infant sex, race, gestational age, delivery method, maternal age, gestational weight gain, insulin sensitivity, total activity counts, basal metabolic rate, respiratory exchange ratio and BDI scores. Sex (P=0.03), race (P=0.02), gestational weight gain (P=0.01), delivery method (P=0.01) and gestational age (P=0.02) were also significantly associated with infant body fat (%) in this study. In conclusion, this longitudinal prospective study provides further support that maternal adiposity is significantly associated with infant body fat at age 2 weeks even after adjusting for important confounders. Ongoing follow‐up of the cohort will determine whether these effects are sustained later in life.

Support or Funding Information

Funded by USDA□ARS Project 6026□51000□010□05S
Young Adult Males and Regional Appalachian Differences in Dietary Patterns across U.S. Universities
Authors: Barr Makenzie L , Charlier Camille M , Colby Sarah E , Greene Geoff , Bryd‐Bredbenner Carol , Olfert Melissa Diann
Published: June 2021

Young adults emerging into independence brings along changes in lifestyle behaviors, specifically dietary patterns. In conjunction with this age point, gender and Appalachian differences emerge. Appalachian regions include portions of New York, Pennsylvania, Ohio, Kentucky, Tennessee, North Carolina, South Carolina, Georgia, Alabama, Mississippi, and all of West Virginia and include populations that tend to be health disparate. Within a sample of 1963 young adults (386 males, 751 females) from 8 universities across the United States, fruit, vegetable, and fat consumption were observed through self‐reported survey data, looking at Appalachian males specifically. The population of males were 53.78% Caucasian, 82.6% non‐Appalachian with a body mass index (BMI) of 24.60±4.32 kg/m2. Females were 59.81% Caucasian, 84.2% non‐Appalachian with a BMI of 24.80±4.26 kg/m2. Using the National Cancer Institute's Fruit and Vegetable Screener (NCI F&V) and Fat Screener (NCI Fat), participants answered survey questions and were scored on the two sections from baseline to 12 months. On a scale of 0–12, with 12 being the highest score of fruit and vegetable consumption, males had an average NCI F&V score of 2.80±0.14 SD per day and females had an average score of 2.22±1.87 SD servings per day. At 12 months, males average serving was 2.28±1.89 SD and females were 2.22+1.87 SD. The averages for NCI F&V were statistically significant across gender (p=0.0005) and time (p=0.218). On a 0–100% scale for NCI Fat, at baseline, males had an average of 30.00±2.66% SD and females had an average of 30.18±2.31% SD. At 12 months, males had an average of 29.35±2.22% SD and females had an average of 29.56±1.83% SD. The results for NCI Fat were also statistically significant across gender (p<0.0001) and approaching it for time (p=0.0664). Post hoc analysis of Appalachian males for each screener indicated no statistical differences across time or in Appalachian for the NCI Fat Screener. However, there was a statistically significant difference in Appalachian males (baseline average = 2.43±2.09 SD, 12 month average = 2.93±2.53 SD) and non‐Appalachian males (baseline average = 2.25±1.85 SD, 12 month average = 2.16±1.81 SD) for their NCI F&V across time (p=0.0072). This large sample of young adult males across the U.S. shows significant differences in gender and residence in Appalachian areas of the U.S. Further analysis of male dietary patterns should be assessed to further aid in tailoring intervention to geographic region.

Support or Funding Information

Approval to use the data set was granted by the University of Tennessee Institutional Review Board prior to study implementation. This material is based upon work that is supported by the National Institute of Food and Agriculture, U.S. Department of Agriculture, under award number 2014‐67001‐21851.
Human Gastrointestinal Microbes Vary Throughout the Day
Authors: Kaczmarek Jennifer L , Holscher Hannah D
Published: June 2021

Background

Human health is intricately intertwined with the composition and function of the trillions of microorganisms that make up the gastrointestinal (GI) microbiota. Habitual diet patterns, rapid changes in dietary fat and fiber composition, and consumption of dietary fibers and prebiotics have all been shown to impact the GI microbiota. Intriguingly, the impact of diet on the microbiota may not only be related to what we eat, but also to when we eat. Emerging research suggests gut microbes experience diurnal rhythms, and that the health effects of eating patterns, such as time‐restricted feeding and meal frequency, may also be related to the GI microbiota.

Objective

We aimed to investigate the relationship between time of day and the composition and function of the gastrointestinal microbiota in healthy adults.

Methods

Healthy adult male and female participants (n=29) provided up to 9 fecal samples (189 total samples) within 15 minutes of defecation over the course of a 15 week trial. Fecal DNA extracts were subjected to 16S ribosomal RNA amplicon‐based high‐throughput sequencing to assess the composition of the microbiota. Sequences were clustered into operational taxonomic units (OTUs) using the Greengenes 13‐8 reference OTU database (97% similarity). Fecal short‐chain fatty acids (SCFA), products of microbial metabolism, were measured by gas chromatography mass spectroscopy. Dietary records were kept throughout the sample collection period and analyzed using the Nutrition Data System for Research (NDSR). Correlations were performed between sample time and bacterial OTUs, and partial correlations between sample time and SCFA concentration were performed with total dietary fiber intake normalized by energy intake as the controlling variable.

Results

Time of defecation ranged from 07:30 to 22:00, median=10:40. Concentrations of acetate, butyrate, and propionate decreased over the course of the day (r=−0.35, −0.31, −0.33, respectively, p<0.0001 for all). The abundance of Clostridium spp. also decreased throughout the day (r=− 0.20, p<0.01). Conversely, Dorea (r=0.18, p=0.01), Oscillospira (r=0.36, p<0.0001), Ruminococcus (r=0.19, p=0.01) and Butyricimonas (r=0.16, p=0.03) increased over the course of the day.

Conclusions

These novel results support and expand preclinical research findings—Oscillospira and Ruminococcus, as well as members of the Lachnospiraceae family and Clostridiales order, have been shown to exhibit diurnal oscillations in rodent models. Our findings also reveal functional changes in the microbiota over the course of the day, as evidenced by reductions in SCFA concentrations with time. In summary, these results suggest that diurnal patterns are also relevant factors for assessments and interventions aimed at modulating the GI microbiota.

Support or Funding Information

Supported in part by the USDA National Institute of Food and Agriculture, Hatch project ILLU‐668‐902.
A Diet Defined by its Content of Bovine Milk Exosomes Alters the Composition of the Intestinal Microbiome in C57BL/6 Mice
Authors: Zhou Fang , Paz Henry A. , Sadri Mahrou , Fernando Samodha C. , Zempleni Janos
Published: June 2021

Background

Exosomes play essential roles in cell‐to‐cell communication. Exosomes and their cargos such as proteins, lipids, and RNAs alter gene expression and metabolism in recipient cells. In previous studies we demonstrated that exosomes are not solely obtained by endogenous synthesis, but that dietary exosomes and their cargos are also bioavailable in non‐bovine species. We further demonstrated that the bioavailability of milk exosomes is less than 100%.

Hypothesis

Dietary intake of exosomes from bovine milk causes changes in the composition of the gut microbiota in mice.

Methods

C57BL/6 mice, age 3 weeks, were fed an exosome‐depleted (E‐) AIN93G‐based diet for up to 42 weeks; controls were fed an exosome‐sufficient (E+) diet. At timed intervals (age 7, 15, 45 weeks), cohorts of mice were euthanized and colon content was flash frozen in liquid nitrogen for subsequent analysis of gut microbiota by 16S rRNA gene sequencing of the V4 region using Illumina's MiSeq platform. Microbial sequences were clustered into Operational taxonomic Units (OTUs). Non‐parametric test was used for statistical analysis.

Results

Depending on sex and age, total 51 OTUs were differentially abundant between treatment groups; See Fig. 1. for a heat map of the top 18 OTUs (P<0.05 for age 15 and 45 weeks). For example, the relative abundance of Firmicute classes Clostridia (Ruminococcaceae) and Verrucomicrobia classes Verrucomicrobiae (Muciniphila) were greater in mice fed E‐compared with E+ at age 15 weeks, whereas the relative abundance of Firmicute classes Clostridia (Clostridiales) was smaller in mice fed E‐ compared with E+ at age 45 weeks.

Conclusions

Consumption of a diet depleted of bovine milk exosomes alters certain microbial taxa in the gut microbiome in mice.

Future plans

Determine whether milk exosomes affect the gut bacterial ecosystem at the functional level and accounts for changes in the host transcriptome and metabolome.

Support or Funding Information

NIFA 2015‐67017‐23181, NIFA 2016‐67001‐25301/NIH DK107264, NIH 1P20GM104320, the Gerber Foundation, and USDA Hatch Act and W3002.
Black Elderberry Extract Improves Serum HDL‐cholesterol and Paraoxonase‐1 Activity in Atherosclerosis‐Prone Mice
Authors: Millar Courtney L. , Winter Heather , Georgelos Jamie , Norris Gregory H , Park Young‐Ki , Blesso Christopher N
Published: June 2021

Cardiovascular disease (CVD) is one of the most prevalent non‐communicable diseases in the United States, claiming upwards of 600,000 lives per year. Excessive plaque accumulation and atherosclerosis are hallmark characteristics of this disease. HDL is responsible for the removal of deposited cholesterol in the arteries to the liver for excretion, a process known as reverse cholesterol transport. Low levels of high‐density lipoprotein cholesterol (HDL‐C) have been correlated with increased risk for CVD in epidemiological studies. However, drug therapies targeting an increase in HDL‐C have failed to demonstrate a reduced risk. More recently, HDL particle functionality rather than HDL‐C concentration, has been suggested to be a better predictor of CVD risk. Ingestion of bioactive dietary components, such as polyphenols, may increase both HDL‐C concentration and HDL functionality. Black elderberry (Sambucus nigra) contains high amounts of anthocyanins (~1300 mg/100 g), a subclass of polyphenols, relative to other edible berries. We investigated whether the consumption of a black elderberry extract (BEE) influenced lipid metabolism, HDL functionality, and atherosclerosis in a mouse model of hypercholesterolemia (apoE−/− mice). We fed male apoE−/− (n = 12/group) ad libitum a low‐fat AIN‐93M diet supplemented with 0 or 1% (by weight of diet; ~150 mg anthocyanins/kg body weight) black elderberry extract (BEE) for 24 weeks. Supplementation with BEE significantly increased total cholesterol (+31.5%) and HDL‐C (+26%), while triglycerides were not significantly changed. Paraoxonase‐1 (PON1), a lipolactonase enzyme carried by HDL which prevents lipoprotein oxidation, was increased by 32% in the treatment group when compared to controls (p < 0.05). Analysis of hepatic gene expression revealed that BEE significantly increased expression of Nrf2 and GClc (p < 0.05), both of which are important for endogenous antioxidant defense and may be related to the significant reductions in serum alanine transaminase and aspartate transaminase levels in the BEE treatment group (p < 0.05). Despite improved markers of HDL‐C and HDL function, en face analysis of aortas showed no difference in severity of atherosclerosis between groups. The current data suggests that BEE supplementation improves HDL‐C and HDL function, while also upregulating genes involved in the endogenous antioxidant defense.

Support or Funding Information

Supported by the USDA National Institute of Food and Agriculture, #1005610 and Hatch project #1003343.