bluuebunny/crossref_metadata_2025 huggingface dataset:
Alternative Splicing Generates Novel Fads3 Transcript in Mice
Authors: Zhang Ji Yao , Qin Xia , Park Hikyu , Kim Ellen , Liu Guowen , Kothapalli Kumar , Brenna J Thomas
Published: June 2021

Introduction

Fatty acid desaturase (FADS) gene family consists of three genes (Fads1, Fads2 and Fads3). Fads1 and Fads2 catalyze critical steps in the biosynthesis of long chain polyunsaturated fatty acids (LCPUFA). Fads3 is the third member, found to have at least eight evolutionarily conserved alternative transcripts (AT), but with no clearly established function. Here we present identification of a novel Fads3 transcript in mice (Fads3AT9). Our main objective is to characterize Fads3AT9, its expression in mouse tissues and in undifferentiated and differentiated fat (white and brown) cells.

Methods.

Total RNA obtained from mouse tissues and fat cells were reverse‐transcribed into cDNA. The resulting cDNA was used as template for PCR‐reactions.

Results.

Sequencing analysis revealed complete absence of exon 2 resulting in an open reading frame of 1239 bp, encoding a putative protein of 412 aa with loss of 37 aa compared to classical Fads3 (Fads3CS). Fads3AT9 retains all the conserved regions characteristic of front end desaturase (cyto b5 domain and three histidine repeats). It is widely expressed in 12 different mouse tissues and fat cells, with strongest expression seen in kidney, and heart. Fads3CS had greater abundance than AT9 in all tissues.

Conclusions

Widespread expression patterns in mouse tissues and fat cells suggest it may play an important role in the regulation and/or biosynthesis of LCPUFA from precursors.

Funding Source:

NIH R01 AT007003
SCD1 Inhibition Modifies Fatty Acid Regulation of Adipocyte Inflammation and Lipid Content
Authors: Ralston Jessica , Metherel Adam , Stark Ken , Mutch David
Published: June 2021

During adipogenesis, adipocytes accumulate triacylglycerols concomitant with increases in the expression and activity of lipogenic genes such as stearoyl‐CoA desaturase 1 (SCD1). SCD1 is the rate‐limiting enzyme for the conversion of dietary and endogenous saturated fatty acids (SFAs) into monounsaturated fatty acids (MUFAs), specifically, palmitate (PA) to palmitoleate (PMA) and stearate (SA) to oleate (OA). Changes in SCD1 activity alter adipocyte SFA/MUFA ratios, which affects cellular function and the development of obesity‐related complications. We examined the bioactivity of 250µM PA, SA, PMA or OA on SCD1‐inhibited adipocytes. SCD1‐inhibited and differentiated 3T3‐L1 adipocytes were treated with fatty acids (FAs) for 48 hrs. FA composition, gene expression, and cytokine secretion were examined by gas chromatography (GC), qRT‐PCR, and multiplex immunoassays, respectively. Treating SCD1‐inhibited adipocytes with PA caused a substantial increase in SA levels that stemmed from an up‐regulation of elongase 6 activity, which was subsequently confirmed using a PA stable isotope tracer and GC coupled mass spectrometry. Adipocytes treated with SFAs showed larger increases in the expression and secretion of markers of inflammation (MCP1, IL6, and CCL5) compared to MUFAs, and these effects were accentuated with SCD1 inhibition. Taken together, reduced SCD1 activity in adipocytes up‐regulates the conversion of PA into SA, and enhances the pro‐inflammatory nature of SFAs.

Grant Funding Source: Supported by NSERC
Attenuation of mTORC1‐Driven Overproduction of ApoB‐Containing Lipoproteins by Aromatic Fatty Acid Phenylbutyric Acid
Authors: Roberts Joseph , Moreau Regis
Published: June 2021

The mechanistic target of rapamycin complex 1 (mTORC1) is a serine/threonine kinase that drives several anabolic processes including lipid synthesis, protein synthesis, and adipogenesis. mTORC1 is highly active in the livers of obese rodents, in overnutrition, and central to the development of lipid abnormalities, including the overproduction of atherogenic lipoproteins. Direct inhibition of mTORC1 is not a suitable therapeutic strategy as it abrogates mTORC1 feedback inhibition of insulin signaling and leads to insulin‐mediated lipid accumulation. An alternative strategy consists in stimulating catabolic pathways to counteract mTORC1‐driven anabolism. Human liver HepG2 cells were stably transduced with lentiviral shRNA targeting tuberous sclerosis complex 2 (TSC2, upstream inhibitor of mTORC1) to constitutively activate mTORC1 or lentiviral scramble shRNA as negative control. In TSC2 shRNA cells, TSC2 gene and protein expression were suppressed by 71% and 75%, respectively, and mTORC1 activity (phospho‐p70 S6K/p70 S6K) was increased by 51% (vs scramble shRNA). The constitutive activation of mTORC1 increased apolipoprotein B (apoB) secretion in culture media by 61% under hyperglycemic (30 mM glucose) conditions. Supplementation with aromatic fatty acid and chemical chaperone, 4‐phenylbutyric acid (8 mM), decreased secreted apoB by 59% compared to vehicle control. We conclude that therapeutic approaches that stabilize improperly folded proteins can reverse the lipoprotein anomalies associated with mTORC1 constitutive activation. Supported by UNL ARD Hatch, and Layman Seed Award.
Characterization and Comparison of Beta‐Lactoglobulin Orthologs
Authors: Crowther Jennifer , Jameson Geoffrey , Hodgkinson Alison , Dobson Renwick
Published: June 2021

Beta‐lactoglobulin (Blg) is the most abundant whey protein in the milk of ruminant animals, yet its physiological function is yet to be confirmed. It is not present in human milk and has been identified as one of the main immunogenic proteins in cow milk. Goat milk appears to be less allergenic and more easily digested than cow milk. As Blg is the most abundant whey protein differences in its structural, physicochemical and dynamic properties may be responsible for some of the differences seen between these milks.

An ultra‐high resolution crystal structure of caprine Blg, along with sedimentation velocity and small‐angle X‐ray scattering data, suggest the presence of subtle differences between bovine and caprine orthologs. While both are dimeric in solution and crystalline states, our data suggest a flexible arrangement of subunits within the dimer. These structural findings have directed further studies into the stability and dynamics of the protein and their binding behaviors with other components in milk, which may provide insight into the differences between these species' milks and potentially the physiological role of this enigmatic protein.
Structural Development of Enzymes Toolbox for Natural Product Biosynthesis
Authors: Wang Fengbin , Singh Shanteri , Helmich Kate , Xu Weijun , Brady Eileen , Huber Tyler , Clinger Jonathan , Bingman Craig , Miller Mitch , Thorson Jon , Phillips George
Published: June 2021

Natural products, known as chemical compounds or substances produced by living organisms, are arguably the most important in revolutionizing modern medicine: penicillin, the aminoglycosides, cephalosporins, tetracyclins and others. To date, natural products remain the best source of drug leads with greater than 50% of the new chemical entities reported since 1981 being derived from natural products. However, one of the shortcomings of natural product‐based drug discovery is the difficulty in structural derivatization. The traditional chemical modification methods remain problematic due to the large variety of functional groups often require multiple protection and deprotection steps, making the process quite difficult and laborious. To help with this, we are trying to make natural product derivatives from enzymatic way. Jon Thorson lab in University of Kentucky has amassed an impressive array of tools and expertise toward understanding and exploiting natural product biosynthesis. Our lab collaborate with them to solve the X‐ray structures of the high‐value, feasible enzyme targets and further engineer them to produce new biosynthetic natural products derivatives. Herein I present structural studies of three parts of our natural product biosynthesis enzymes toolbox: (1) methyltransferases (MTs) and methionine adenosyltransferases (MATs); (2) glycosyltransferase (GTs); (3) sugar aminotransferase.
Investigating the Mechanism of Action of Sulforaphane in a Human Leukemia Cell Line
Authors: Fields Peter , Grom Jennifer , O'Donnell Robert
Published: June 2021

Sulforaphane is an isothiocyanate molecule that is found in cruciferous vegetables, such as broccoli. Its anti‐neoplastic properties make it useful as a chemotherapeutic drug. Although it is known to be an effective treatment against leukemia tumor cells, the mechanism by which sulforaphane kills leukemia cells is unknown. Previous labs have shown that sulforaphane effectively arrests leukemia cells in the G2/M cell cycle phase, but what is not clear is whether the phase is G2 or M. In order to clarify the affected phase, roscovitine will be used as a G2 inhibitor and nocodazole will be used as an M phase inhibitor. HL60 leukemia tumor cells will be treated with sulforaphane, roscovitine, and nocodazole, and the results will be compared. Preliminary experiments used sulforaphane, roscovitine, and nocodazole MTT assays to determine the concentration resulting in 75% survival. The concentrations were found to be 4.71 μM, 16.21 μM, and 26.40 nM, respectively. Future experiments will use these doses to compare and identify where the inhibition occurs. Western Blots will be used to look at p21, an inhibitor of several CDK/cyclin complexes, and cdc2 levels (the CDK required to initiate the G2 to M phase transition). Mitotic index and cell cycle analysis will also be used. We hypothesize that if we treat the HL‐60 cells with sulforaphane, there will be an increase in the p21 levels of the cells, supporting arrest in the G2 phase of the cell cycle. We also hypothesize that treatment of leukemia tumor cells with sulforaphane or roscovitine will yield low levels of cdc2 (compared to untreated cells), while nocodazole treated cells will have cdc2 levels that are similar to the untreated cells.
Rumex crispus Phytochemicals: Chemopreventative Potential for Human Colon Carcinoma
Authors: Lee Dallas , Clack Beatrice
Published: June 2021

The study and isolation of naturally occurring phytochemicals has produced several chemotherapeutic drugs and derivatives still in use today including: paclitaxel (taxol), etoposides (podophyllotoxin), and irinotecan. The objective of this study was to isolate novel chemotherapeutic compounds of Rumex crispus (R. crispus) using high performance liquid chromatography (HPLC), characterize with mass spectrometry (MS), and test with in vitro assays.

Human colon adenocarcinoma cells (DLD‐1ATCC® CCL‐221™) were used to test the effectiveness of R. crispus phytochemicals as chemotherapeutics. Accelerated solvent extraction (ASE) was utilized to extract 0.1% TritonX‐100 soluble phytochemicals from R. crispus obtained in East Texas. Extracts from both rhizomes and leaves were screened for chemotherapeutic activity using CellTiter 96® Non‐Radioactive Cell Proliferation Assay (Promega, Corp.) and the Apo‐ONE® Homogeneous Caspase‐3/7 Assay (Promega, Corp.). To further explore the relative types of phytochemicals responsible for cytotoxicity and apoptosis in the cells, reverse phase HPLC and MS was utilized. Phytochemical fractions collected from HPLC were titrated with NaOH, to neutralize the H3PO4 present from HPLC. Each HPLC fraction was tested for levels of cytoxicity using the CellTiter 96® Non‐Radioactive Cell Proliferation Assay (Promega, Corp.). Those fractions with positive killing were then subjected to MS to identify specific phytochemicals present.

Cytotoxic effects were observed in both rhizome and leaf extracts of R. cripsus, as well as the induction of apoptosis with increasing dosage of R. crispus extracts. Large amounts of benzylphytosterol or benzylisoquinalone type molecules were present in the MS data of ASE R.crispus root extracts.
The National Cancer Institute's Dietary Assessment Primer
Authors: Krebs‐Smith SM , Kirkpatrick SI , Subar A F , Rodgers A B , Schap TE , Reedy J , Wilson MM , De Aguiar CK , Thompson FE
Published: June 2021

The efficacy of all dietary studies, whether aimed at monitoring a population's diet, understanding diet and health relationships, or evaluating the effect of an intervention, depends on appropriate assessment of intakes. Determining the most suitable dietary instrument for a particular study can be a challenge and is dependent upon a number of considerations, including the specific research question, dietary components of interest, study design, and target population. The National Cancer Institute has developed an online resource to provide guidance on the selection of appropriate dietary assessment methodologies for characterizing the intakes of a group or groups and to outline various considerations for the use of each. The Dietary Assessment Primer describes the major types of self‐report instruments; provides guidance on using the instruments alone or in combination to address different research questions; compares key features of each instrument; explains validity, measurement error, and calibration in the context of dietary assessment; provides expanded information about particular key topics; and includes a Glossary of basic terms and an extensive list of References and Resources. The online tool was developed by subject matter experts within NCI, with internal and external review to ensure the content reflects the current state of the science. User‐centered design principles were applied to developing the web‐based interface to ensure optimal usability. The Primer is intended to help researchers collect the highest quality dietary data possible given their study resources and constraints, with the ultimate goal of improving our capacity to monitor diets among populations and understand how diet affects health.
Breakfast Eating and Incident Coronary Heart Disease in a Large Prospective Cohort of American women
Authors: Cahill Leah , Chiuve Stephanie , Mekary Rania , Jensen Majken , Flint Alan , Hu Frank , Mukamal Kenneth , Rexrode Kathryn , Rimm Eric
Published: June 2021

Background

The 2010 Dietary Guidelines for Americans recommend breakfast for children but found insufficient evidence to make recommendations for adult eating habits (the timing and frequency of meals, snacks, and caloric beverages).

Objective

To conduct the first long‐term prospective study of eating habits and risk of incident coronary heart disease (CHD) in women.

Methods

Eating habits including breakfast eating and eating frequency were assessed in 2002 in 56,593 women from the Nurses' Health Study who were free of cardiovascular disease and cancer. During 8 years of follow‐up, 770 incident CHD cases were diagnosed. Cox proportional hazards models were used to estimate relative risks (RR) and 95% confidence intervals (CI) for CHD, adjusted for demographics, diet, lifestyle, and other CHD risk factors.

Results

Among women less than 60 years old, those who skipped breakfast regularly had a higher risk of CHD (RR=3.82, 95% CI:1.07‐13.67) as compared to women who ate breakfast at least 5 times/week. However, this association was not present among women 60 years or over (RR=0.98, 95% CI:0.74‐1.30) (p, interaction = 0.05). No associations were observed between eating frequency (times/day) and risk of CHD.

Conclusion

Eating breakfast was associated with a lower CHD risk in women under 60 years of age, but not in older women.

Support

NIH Grants CA176726, HL034594
Metagenomic Systems Biology: Metabolic Modeling and Multi‐Meta‐Omic Analysis of the Human Microbiome
Authors: Borenstein Elhanan
Published: June 2021

The human microbiome represents a vastly complex ecosystem that is tightly linked to many host‐related processes and that directly impact human health. To date, however, most studies have focused on characterizing the composition of the microbiome in health and in disease and on comparative analyses, and relatively little effort has been directed at studying and modeling the microbiome as an integrated and comprehensive biological system. In this talk, I will highlight the pressing need for the development of predictive system‐level models of the microbiome and discuss potential computational frameworks for metagenomic‐based modeling of this microbial system. I will describe several preliminary attempts at constructing such models at the cellular, ecological and supra‐organismal levels, accounting for the complex web of interactions in the microbiome. Specifically, I will demonstrate the use of network‐based and constraint‐based models to study various aspects of the microbiome, predicting both microbe‐microbe and microbe‐host interactions. These models allow us to obtain valuable insights into the forces that govern the assembly of the microbiome and into disease‐associated organizational shifts. Finally, I will discuss routes for integrating various modeling approaches and multiple meta‐omic datasets for obtaining a comprehensive, multi‐scale, mechanistic understanding of the microbiome and exciting future clinical applications facilitated by this research.
Piloting of a Learner‐Centered Nutrition Curriculum for School Nutrition Staff
Authors: Jones Anna , Brian K. , Briggs M. , Zidenberg‐Cherr S.
Published: June 2021

The purpose of this study was to pilot a nutrition education curriculum for school nutrition staff. Backwards design was utilized to develop lessons centered around learning objectives and key concepts. The curriculum consisted of five lessons about concepts such as nutrients in foods, portion sizes, and physical activity, with applications to the National School Lunch Program throughout. It was piloted in Aug. 2014, in 2 one‐day workshops in 2 school districts in Northern California. The workshops were evaluated qualitatively using a questionnaire completed at the conclusion of each workshop (n=46). The questionnaire results were analyzed for common themes. The curriculum was well received by participants; over 40% of respondents specifically stated the information provided was the most valuable part of the workshop. In particular, participants indicated they learned the most about nutrients in foods (23%) and reading nutrition facts labels (20%). Participants also appreciated the learner‐centered design, with 30% stating that was what they liked best and over 90% agreeing that the course delivery was effective. Improvements to the course were identified; 7% of participants expressed that the course was repetitive at times, while others did not find the lesson on nutrition in the media to be useful (15%). Overall, the results demonstrate that this curriculum has potential utility; further research should be conducted to evaluate effectiveness at improving nutrition knowledge and self‐efficacy. Funding was provided in part by the California Department of Education.
Changes in the Intestinal Microbiota and Host Inflammatory Gene Expression in Pigs Fed a Flavanol‐Enriched Cocoa Powder
Authors: Jang Saebyeol , Sun Jianghao , Chen Pei , Lakshman Sukla , Molokin Aleksey , Harnly James , Urban Joseph , Davis Cindy , Solano‐Aguilar Gloria
Published: June 2021

To determine the effects of eating cocoa‐derived flavanols on the composition of the gut microbiota, associated metabolic profiles, and related features of host intestinal health, pigs were fed with 0, 2.5, 10, or 20 grams of flavanol‐enriched cocoa powder/day for 27 days. Analysis of phase II metabolites of cocoa‐derived flavanols demonstrated that O‐methyl‐(epi)catechin‐glucuronide conjugates were detected in a dose‐dependent manner in urine, serum, liver, and adipose tissue of pigs fed cocoa powder. Products of microbial metabolism such as phenylpropionic acids, phenylacetic acids, benzoic acids, cinnamic acids, and conjugated benzoic acids were detected in most of the tissue and bio‐fluids examined. Cocoa consumption also changed the composition of the intestinal microbiome. Pigs fed 20 gram of cocoa/day had significantly increased abundance of Lactobacillus species (casei group) in the feces and distal colon contents compared to pigs not fed cocoa. Moreover, consumption of cocoa powder reduced the gene expression of tumor necrosis factor‐α and toll‐like receptors‐2, ‐4, and ‐9 in most intestinal tissues examined. Overall, this study demonstrated that consumption of flavanol‐enriched cocoa can contribute to gut health by enhancing the abundance of Lactobacillus species and modulating markers of localized intestinal immunity.
Effect of glutathione S‐transferase (GST) M1 and T1 polymorphisms on lymphocyte DNA damage of Korean subclinical hypertensive patients after kale juice supplementation
Authors: Kang Myung‐Hee , Han Jeong‐Hwa , Lee Hye‐Jin , Kim Tae‐Seok
Published: June 2021

Glutathione S‐transferase (GST) forms a multigene family of phase II detoxification enzymes that are involved in the detoxification of xenobiotics by conjugating substances with glutathione. In this study, we examined whether supplementation of kale juice could modulate the levels of lymphocyte DNA damage in hypertensive patients, and whether this modulation could be affected by GSTM1 and T1 polymorphisms. Eighty‐four subclinical hypertensive patients showing systolic blood pressure (BP) over 130 mmHg or diastolic BP over 85 mmHg received 300 ml/day of kale juice for 6 weeks, and blood samples were drawn before and after the 6 weeks. DNA was extracted from whole blood samples, and GSTM1 and GSTT1 genotypes were determined using a multiplex PCR. Lymphocyte DNA damage was measured by comet assay and damage was quantified by measuring tail moment, tail length (μm), and DNA in tail (%). The level of DNA damage decreased significantly in both GSTM1‐null and GSTM1‐present genotypes (p<0.001) and decreased in both GSTT1‐null and GSTT1‐present genotypes after supplementation (p<0.001); however, after the 6‐week supplementation, the reduced effect of DNA damage was significantly greater in the GSTM1‐null genotype compared with the GSTM1‐present genotype. These results suggest that supplementation of kale juice might prevent lymphocyte DNA damage in subclinical hypertensive patients, and this preventive effect is affected by GSTM1 genotype (This research was supported by the Basic Science Research Program of the National Research Foundation of Korea (NRF) under the sponsorship of the Ministry of Education, Science and Technology (2010‐0012031)).
Cornus officinalis Modulates the Production of Pro‐Inflammatory Molecules in Lipopolysaccharide‐Activated RAW264.7 Macrophages
Authors: Akhavan Neda , Feresin Rafaela , Johnson Sarah , Pourafshar Shirin , Elam Marcus , Hsieh Yun‐Hwa , Salazar Gloria , Arjmandi Bahram
Published: June 2021

Cornus officinalis (CO) are berries found predominantly in China and Korea and have been used in Eastern medicine due to their polyphenolic content. The objective of this study was to investigate the inhibitory effects of different doses of CO extract (COE) on inflammatory molecules including nitric oxide (NO), cyclooxygenase‐1 (COX‐1), cyclooxygenase‐2 (COX‐2), and tumor necrosis factor‐alpha (TNF‐α) in lipopolysaccharide (LPS)‐activated RAW264.7 macrophages. CO was extracted using methanol and total polyphenolic content (TPC) was determined using high performance liquid chromatography. Macrophages were treated with different doses of COE (0, 50, 100, 200, 400 µg/mL) two hours prior to stimulation with 100 ng/mL of LPS. TPC of COE was 1,872.8 ppm with anthocyanins and chlorogenic acid being the most abundant polyphenols found in COE. COE at doses of 50, 100 and 200 µg/ml were able to significantly (P < 0.05) decrease NO levels compared to LPS. No significant changes were observed in TNF‐α levels. COE dose‐dependently decreased COX‐2 levels when compared to LPS (P < 0.05) while no effects of COE were observed on COX‐1 levels. The results from study indicate that COE is rich in polyphenols and possesses anti‐inflammatory properties suggesting a possible role as a complementary and alternative therapy to prevent inflammation. However, in vivo and human studies are needed to confirm this.
Blueberry Bioactives Improve Endothelial Function in Adults with Metabolic Syndrome
Authors: Stull April , Cash Katherine , Champagne Catherine , Gupta Alok , Boston Raymond , Beyl Robbie , Johnson William , Cefalu William
Published: June 2021

There is a paucity of data suggesting that increased blueberry consumption improves cardiovascular risk factors in human subjects. This study investigated the role of blueberry bioactives on modifying vascular function and blood pressure in subjects at high risk for developing cardiovascular disease. A randomized, double blinded, and placebo‐controlled study was conducted in 44 adults with metabolic syndrome. They were randomized to receive twice daily blueberry bioactives (22.5g of freeze‐dried whole blueberry powder) added to a smoothie or an identical smoothie without the blueberry bioactives (i.e., placebo) per day for 6 weeks. Endothelial function (peripheral arterial tone) and blood pressure (7‐day automatic blood pressure monitoring device) were assessed pre‐ and post‐intervention. The subjects' mean change in blood pressure did not differ between the blueberry and placebo groups. However, the mean change in resting endothelial function, expressed as reactive hyperemia index, was improved significantly more in the blueberry versus placebo groups (P<0.05). Even after adjusting for confounding factors, i.e., percent body fat and gender, the blueberry group still had a greater improvement in endothelial function when compared to their counterpart. In conclusion, daily dietary supplementation of blueberry bioactives improved endothelial function over 6 weeks in subjects with metabolic syndrome.
Effects of Watermelon Powder on Lipid Metabolism through Regulation of Gene Expression in Atherogenic Diet‐Fed Rats
Authors: Kaufman Katy , Hooshmand Shirin , Kern Mark , Figueroa Arturo , Hong Mee Young
Published: June 2021

Cardiovascular disease (CVD) is the leading cause of death in the United States, which has led to an increasing need for cost effective non‐pharmacological treatment alternatives. Studies have shown that watermelon plays a role in vascular function by causing vasodilation, which lowers blood pressure and arterial stiffness. We want to examine if watermelon powder supplementation has therapeutic effects on lipid metabolism through regulation of hepatic gene expression. The genes of interest included fatty acid synthase (FAS), HMG‐CoA reductase (HMGCR), sterol regulatory element binding protein ‐1 and 2 (SREBP‐1, SREBP‐2). Forty male‐weanling (21 days old) Sprague‐Dawley rats were divided into four groups (10/group, total N=40) in a 2 (diets) x 2 (treatment) factorial design using an atherogenic diet with or without Dextran Sodium Sulfate (DSS) as an inflammatory agent for 48 hours. Watermelon powder groups exhibited significantly lower total cholesterol levels, LDL‐cholesterol, and serum triglycerides (P<.05). FAS, HMGCR, SREBP‐1, and SREBP‐2 were significantly down regulated in watermelon diet groups (P<.05). These findings support the use of watermelon powder as an alternative treatment for improving risk factors associated with CVD by altering hepatic lipid metabolism related gene expression and thus improving lipid profiles. Supported by NUTR 302L class and SDSU UGP.
In vitro Evaluation of Wild Fruits and Leaves for Potential Carbohydrate Hydrolyzing Enzyme Inhibition
Authors: Lee Diana , Racicot Kenneth , Apostolidis Emmanouil
Published: June 2021

Four wild foods dandelion leaf ‐ (Taraxacam officinale), grape (Vitis riparia) plantain (Plantago major), and beach plum (Prunus maritima) ‐, were extracted in water followed by C18 column extraction. The phenolic content was evaluated using the Folin‐Ciocalteau's Assay and determined to be 39.5 μg/mg GAEq, 116.2 μg/mg GAEq, 14.3 μg/mg GAEq, and 351.1 μg/mg GAEq for the dandelion, grape, plantain, and plum respectively. The antioxidant activity was evaluated in terms of free‐radical scavenging activity, by using the DPPH assay. Single dose evaluation showed that the sample with the highest free‐radical scavenging activity was the beach plum with 70.2% antioxidant activity, followed by grape with (58.6%), dandelion (50.0%), and plantain (44.9%). Dose‐dependent evaluation of free‐radical scavenging activity is currently underway. Previous reports have shown that phenolic phytochemicals are excellent inhibitors of carbohydrate hydrolyzing enzymes. To determine the possible effect of our samples for type 2 diabetes prevention, we will evaluate their dose‐dependent inhibitory effect against two major carbohydrate hydrolyzing enzymes (alpha‐glucosidase and alpha‐amylase). Completion of this research will provide the basic biochemical rationale for the deeper evaluation of samples with high inhibitory activity.
Xylooligosaccharide Induced Changes in Gut Microbiota in Healthy and Prediabetic Adults
Authors: Li Zhaoping , Yang Jieping , Carslon Paula , Henning Susanne , Hsu Mark , Tseng Chi‐Hong , Thames Gail , Finegold Sydney , Heber David
Published: June 2021

Background

It has been shown that the abundance and diversity of the gut microbiota is altered in individuals with metabolic disorders.

Objective

This study was conducted to evaluate the effect of prebiotics xylooligosaccharide (XOS) on gut microbiota and glucose metabolism in healthy and prediabetic subjects.

Design

Prediabetic (n=18) or healthy (n=16) adult subjects were randomized to receive 2.8 g/day XOS or placebo in an 8‐week study. In prediabetic subjects, blood and stool were collected, and body composition and oral glucose tolerance test (OGTT) was done at baseline and after 8 weeks of intervention. Stool from prediabetic (n=13) and healthy (n=16) subjects at baseline and week 8 was processed for gut microbiota characterization using Illumina MiSeq sequencing.

Results

XOS induced notable similar changes in microbial diversity in both healthy and prediabetic subjects, as well as changes varied by the subject health status. The baseline increase of genera Enterorhabdus was seen in prediabetes compared to healthy subjects. XOS significantly reverse this transformation in prediabetes. The abundance of the class Coriobacteria was reduced by XOS in both healthy and prediabetic adults. Genera Dialister was only increased by XOS in prediabetic subjects. The increased of Dialister was reported in people receiving dietary whole grain intervention. XOS lowered 0‐2h OGTT insulin response (P=0.11), but had no effect on body composition, serum glucose, triglyceride, satiety hormone and inflammation markers in prediabetic subjects.

Conclusions

Further studies are needed to determine the role of Enterorhabdus, Coriobacteria and Dialister in the use of prebiotics in prediabetes.
NMR‐Based Metabolomic Profiles of Mice Fed a High Fat Diet and Supplemented with Resistant Starch and/or Lactobacillus
Authors: Martinic Alice , Mishchuk Darya , Berouei Javad , Kieffer Dorothy , Martin Roy , Marco Maria , Slupsky Carolyn
Published: June 2021

Incidence of obesity and type 2 diabetes mellitus (T2DM) is steadily increasing, and the contributions of diet and the gastrointestinal microbiota to these diseases are only partially understood. Enriching particular populations of microbes in a mouse model of pre‐diabetes and monitoring systemic metabolic effects will help to elucidate the mechanisms of obesity and T2DM progression. C57BL/6J mice were fed a 45% high fat diet for nine weeks and then supplemented with type 2 resistant starch (RS), Lactobacillus plantarum (Lp), or both for six weeks (n=10 per group). Urine was collected throughout the experiment; cecal contents and serum were obtained at the time of mouse sacrifice. All samples were analyzed using NMR‐based metabolomics. Unsupervised principal component analysis showed clear separation between RS and non‐RS groups for both cecal and serum samples. Significant differences were observed for microbial metabolites and amino acids when comparing RS groups to control, as well as when comparing Lp supplementation alone to RS supplementation alone. These results have implications for monitoring the effects of dietary supplementation on the gastrointestinal microbiota and overall host health. Funding was provided by the American Diabetes Association.
Raisins (Vitis vinifera L.) as a Source of Prebiotic Functional Food
Authors: Mandalari Giuseppina , Ianson Kerry , Chan Luisa , Carughi Arianna
Published: June 2021

In the present work we report the potential prebiotic effect of sun‐dried raisins, known to be a source of fructo‐oligosaccharides (FOS). A full model of gastrointestinal tract digestion, including gastric and small intestinal environments and a colonic model of in vitro fermentation with representative human gut bacteria was used. Bacterial identification was performed by plate counting on selective media and 16S V4 rDNA analysis.

Results from plate counting showed an increase in the numbers of beneficial bacteria, such as bifidobacteria and lactobacilli, after fermentation with raisins. Higher prebiotic index (PI) scores were recorded after 8 and 24 h fermentation with raisins compared with negative control (no carbon source added). Microbial profiling showed a bacterial abundance shift in the whole microbiome, with an increase of Bifidobacteria after fermentation with raisins and FOS, used as positive control. This trend was stronger with Bifidobacterium sp. than Lactobacillus sp. The concentration of lactic and acetic acids increased after fermentation with raisins (11.6 and 36.5 mmol/l after 24h, respectively), demonstrating a beneficial effect correlated to the increase in the number of beneficial bacteria.

In conclusion, we have shown that bioactive components in raisins were able to modulate the growth of gut bacteria. Sun‐dried raisins exhibited the potential to be used as a novel source of prebiotics.
An In Vitro and In Vivo study of BNAP: A Novel Peripherally Selective μ‐opioid Receptor Antagonist
Authors: Williams D , Yuan Y , Stevens D , Zhang Y , Dewey W , Akbarali H
Published: June 2021

Activation of μ‐opioid receptors (MORs) on enteric neurons during opioid treatment inhibits gastrointestinal peristalsis and leads to constipation. This is the most significant side affect associated with opioid treatment. Peripherally active MOR antagonists can minimize constipation without affecting analgesia. Previously, we described the 6β‐Naltrexamine derivative NAP as a novel peripherally selective MOR antagonist. NAP was 300x more potent than methylnaltrexone at improving intestinal motility in morphine tolerant mice. However, NAP partially antagonized morphine antinociception in the mouse tail‐flick assay. Efforts to reduce CNS activity led to the development of BNAP. Radioligand binding studies show BNAP maintained high MOR affinity (K­i = 0.76 ± 0.09 nM). In the mouse tail flick assay BNAP (10mg/kg) did not antagonize morphine‐induced antinociception. BNAP also competitively inhibited morphine‐induced contractions in circular muscle preparations from the mouse distal and proximal colon. Both NAP and BNAP antagonized morphine‐induced contractions with equal efficacy at the doses tested (1, 10, and 100 nM). Interestingly, they were 10x more potent in the proximal colon. This suggests differences in MOR receptor density or splice variants between proximal and distal colon. In conclusion, BNAP is a more peripherally selective MOR antagonist than NAP and has equal potency in isolated tissue preparations. Thus, BNAP is a suitable lead for further study and development as a novel peripherally selective MOR antagonist. Supported by DA024009, DA024002, and DA007027.
Gating Effects of a Novel Allosteric Modulator at GluN1/GluN2A NMDA Receptors
Authors: Chopra Divyan , Irvine Mark , Jane David , Monaghan Daniel , Dravid Shashank
Published: June 2021

NMDA receptors are members of the ionotropic glutamate receptor family and are critical for processes such as synaptic plasticity. They also represent important therapeutic targets for CNS disorders. However only a few NMDA receptor agents have successfully transitioned to clinical setting, thus there is a need to develop novel modulators preferably with different sites and mechanism of action which limit potential side effects. NMDA receptors are tetramers usually composed of two GluN1 and two GluN2 subunits. The goal of the current study was to evaluate the gating mechanisms of a novel allosteric modulator at the GluN1/GluN2A NMDA receptor. We used whole‐cell and cell‐attached patch recordings to evaluate the gating mechanism of a UBP series compound. In whole‐cell recordings we found that UBP684 increased the steady state current by 84.16±18%.We further performed cell‐attached recordings and observed that UBP684 increased the mean open time: control=1.57±0.53ms and UBP684=4.34±2.14ms and the open probability: control=0.08±0.02 and UBP684=0.61±0.15 and also reduced the mean shut times.The open time and shut time histograms were fitted with 3 and 5 exponential components respectively. UBP684 was found to primarily affect the area of the time constants and not the time constants themselves. Kinetic modeling suggests that UBP684 has a profound effect on one of the slower gating steps. Our studies indicate that UBP684 may have subunit‐selective effects and provides insight into a novel potentiating mechanism for GluN1/GluN2A NMDA receptors.
CYP2A6 Genetic Variation and Variable Nicotine Metabolism Among Two Distinct American Indian Tribal Groups With Different Levels of Smoking and Risk For Tobacco‐Related Cancer
Authors: Tanner Julie‐Anne , Henderson Jeffrey , Howard Barbara , Buchwald Dedra , Tyndale Rachel
Published: June 2021

CYP2A6 variability, resulting in altered rates of nicotine metabolic inactivation, is associated with variation in smoking behaviors. Distinct patterns of smoking and disease prevalence of two American Indian (AI) tribes (Northern Plains, NP; Southwestern, SW) prompted our investigation of CYP2A6 genetic variability and nicotine metabolism in NP and SW AIs. Both tribes (NP n=426; SW n=210) were genotyped for multiple CYP2A6 variants representative of prevalent loss of function alleles from different ethnicities. Using 3'‐hydroxycotinine to cotinine (3HC/COT) ratio as a phenotype of nicotine metabolism, association between CYP2A6 genotype and rate of nicotine metabolism was examined. CYP2A6 genotype was associated with the rate of nicotine metabolism in both tribes (P<0.02), confirming that CYP2A6 genotype predicts rate of nicotine metabolism for these populations. The rate of nicotine metabolism was higher in NP compared to SW AIs when controlling for genotype (P<0.01). The frequency of reduced metabolizers, those with loss of function alleles, was higher in the SW compared to the NP tribe (P<0.01). Faster nicotine metabolism, and fewer reduced function variants, which was observed in the NP tribe, has been associated with higher levels of smoking and dependence, more difficulty quitting, and reduced response to some cessation pharmacotherapies in other populations.

Funding: CIHR and the Collaborative to Improve Native Cancer Outcomes (NCI grant P50CA148110)
UM206, a Peptide Fragment of Wnt5a, Attenuates Adverse Remodeling after Myocardial Infarction in Swine
Authors: Hermans Kevin , Uitterdijk André , de Wijs‐Meijler Daphne , Daskalopoulos Evangelos , Verzijl Annemarie , Sneep Stefan , Blonden Lau , Reiss Irwin , Duncker Dirk , Blankesteijn W Matthijs , Merkus Daphne
Published: June 2021

Introduction

Following myocardial infarction (MI), left ventricular (LV) remodeling aims to maintain cardiac function. Nevertheless, adverse remodeling results in thinning of the infarct and LV dilation which may subsequently lead to the development of heart failure (HF) with poor prognosis. Wnt/Frizzled signaling plays a key role in cardiac remodeling following MI. We have shown previously that blockade of Wnt/Frizzled signaling with UM206 suppresses adverse remodeling and prevents HF development in mice post‐MI. Here we investigated the effects of UM206 therapy in a clinically relevant large animal model.

Methods

Twelve Yorkshire x Landrace swine were subjected to 2h of myocardial ischemia by ligation of the proximal left circumflex artery, followed by reperfusion. Starting 24h post‐MI, 6 animals were treated with UM206 (0.6 μg/kg/day i.v.) for 5 weeks. The 6 control swine were treated with vehicle. Cardiac dimensions and infarct mass (IM) were determined by echocardiography and dedicated markers, respectively.

Results

Treatment with UM206 for 5 weeks resulted in a significant decrease in IM compared to baseline, whereas IM remained unaffected in the control group. This was accompanied by progressive dilation of the LV in the control group between 3 and 5 weeks after MI while adverse remodeling was halted in the UM206 treated group.

Conclusion

UM206 treatment in a clinically relevant swine model of reperfused myocardial infarction attenuates adverse remodeling.

Supported by the Dutch Heart Foundation (2010B196)
Effect of thoracic epidural anesthesia on heart rate variability in a porcine model
Authors: Mahajan Arjun , Pakanati Varun , Yamakawa Kentaro , Zhou Wei
Published: June 2021

Heart rate variability (HRV) is increasingly recognized as a means of evaluating cardiac autonomic tone. Our group previously has demonstrated that thoracic epidural anesthesia (TEA) successfully suppressed the electrical storms in patients. However the effect of TEA on HRV during sympathoexcitation remains unknown. Our objective of this study was to determine the effects of stellate ganglion (SG) stimulation on HRV by using power spectral analysis as well as the effects of TEA on HRV response to SG stimulation. After insertion of an epidural catheter from T7/8 placing distal tip of the catheter to T1 level, a median sternotomy was performed to expose the heart and the bilateral SG. A 56‐electrode sock was used for obtaining epicardial activation recovery interval (ARI), a surrogate marker of action potential duration. Fifteen anesthetized pigs received SG stimulation at 5 Hz for 30 second. SG stimulation significantly increased low frequency normalized units (LF n.u.) component (42.7±9 vs. 12.4±3 baseline, p<0.05) indicating enhanced sympathetic nerve activity and decreased high frequency normalized units (HF n.u.) component (26.5±3.2 vs. 56.4±10 baseline, P<0.05) indicating a decreased parasympathetic nerve activity. As a result, LF/HF significantly increased from 0.22 to 1.61 during SG stimulation. LF component significantly correlates with ARI (r=0.81) and ARI dispersions (r=0.85). TEA significantly attenuated the LF/HF from 1.6 to 0.6 with increased HF components from 26.5±3.5 to 48.2±4.1(n.u.). In conclusion, HRV can precisely reflect the cardiac autonomic tone and TEA modulates the HRV by enhancing the HF components.

LF (n.u.)
HF (n.u.)
LF/HF

Baseline
12.4±3.2
56.4±10.1
0.22±0.1

SG stim
42.7±8.5
26.5±3.2
1.6±1.0

SG stim + TEA
37.7±6.2
48.2±4.5
0.78±0.1
Enhanced External Counterpulsation Reduces Indices of Left Ventricular Wasted Energy and Myocardial Oxygen Demand in Patients with Left Ventricular Dysfunction and Refractory Angina
Authors: Beck Darren , Martin Jeffrey , Casey Darren , Sardina Paloma , Braith Randy
Published: June 2021

Enhanced external counterpulsation (EECP) therapy decreases angina episodes and improves quality of life in patients with left ventricular dysfunction (LVD). However, the underlying mechanisms of EECP therapy in patients with LVD have not been fully elucidated. The purpose of this study was to investigate the effects of EECP on indices of central hemodynamics, aortic pressure wave reflection characteristics and estimates of LV load and myocardial oxygen demand in patients with LVD. Patients with chronic stable angina and left ventricular ejection fraction (LVEF) <40%, but >30%, were randomized to either an EECP (LVEF=35.1±4.6%; n=10) or sham‐EECP (LVEF=34.3±4.2%; n=7) group. Pulse wave analysis of the central aortic pressure waveform and LV function were evaluated by applanation tonometry before and after 35 1‐hr sessions of EECP or Sham EECP. EECP therapy was effective in reducing indices of left ventricular wasted energy and myocardial oxygen demand by 25% and 19%, respectively. In addition, indices of coronary perfusion pressure and subendocardial perfusion were increased by 9% and 30% after EECP, respectively. Our data indicate that EECP may be useful as adjuvant therapy in heart failure patients through reductions in central blood pressure, aortic pulse pressure, wasted left ventricular energy, and myocardial oxygen demand which suggests improvements in ventricular‐vascular interactions.
High Blood Pressure Induced by Vitamin D Deficiency (VDD) is Associated with Upregulation of NKCC2 and Intrarenal RAS Activation
Authors: Luchi Weverton , Crajoinas Renato , Seguro Antonio , Girardi Adriana
Published: June 2021

VDD has been linked to hypertension in experimental and clinical studies. Accordingly, we have recently found that Wistar rats fed a vitamin D‐free diet for 30 days (VDD30) display higher mean blood pressure compared to controls fed standard diet (142±2 vs. 117±3 mmHg, p<0.001). Thus, we aimed to test the hypothesis that high blood pressure in VDD30 rats is associated with altered expression of apical sodium transporters along the nephron and intrarenal RAS activation. Compared to controls, VDD30 rats had lower serum levels of 25(OH)D and similar plasma calcium and phosphorus. Urinary flow and fractional sodium excretion were not affected after 30‐day VDD, however it slightly reduced GFR. Immunoblotting analysis showed that VDD30 rats exhibit higher expression of NKCC2 in medullary (147±12 vs. 100±10%, p<0.05) and cortical (157±12 vs. 100±10%, p<0.05) kidney compared to controls. No significant changes were observed on the renal cortical levels of NHE3, NCC, NCC phosphorylated at Thr 53 and in cortical and medullary α, β and γ ENaC subunits between groups. However, the levels of NHE3 phosphorylated at Ser 552 was higher in the renal cortex of VDD30 rats (200±12 vs. 100±11%, p<0.001), possibly as part of the pressure‐natriuresis response. Renin expression did not vary in the renal cortex, but was markedly elevated in the medulla of the VDD30 rats (207±28 vs. 100±25%, p<0.05). Higher expression of cortical angiotensinogen was found in VDD30 rats (160±18 vs. 100±19%, p<0.05). On the other hand, plasma angiotensinogen was similar to control rats. Collectively, these data suggest that VDD leads to altered renal sodium handling and high blood pressure by upregulating NKCC2 and intrarenal RAS.
Exercise training prevents hypertension, dyslipidemia and renal lipid deposition in young rats fed a high‐fat diet
Authors: Muller Cynthia , Americo Anna Laura , Neto Giuseppe , Evangelista Fabiana , Farah Vera , Fiorino Patricia
Published: June 2021

We investigated the potential of exercise training (ET) to prevent hypertension, lipid profile damage and intrarenal lipid deposition in young rats. Male Wistar rats (n=6/groups) were separated into sedentary (SC) or trained (TC) fed a control diet, and sedentary (SH) or trained (TH) fed a high‐fat diet (30% lipids) during 12 weeks since the weaning. ET was performed in treadmill (60% of maximal velocity, 1h/day, 5x/week, during 10 weeks) and started in the second week of the protocol. After ET, the animals were catheterized to record basal blood pressure (BP) and heart rate (HR) during 15 minutes. Then, the animals were killed by overload of anesthetic (ketamine/Xylazine, ip.) and left kidney was harvested to quantify the lipid deposition by oil red staining. Serum was collected to evaluate lipid profile by colorimetric kit. Basal BP was 10% higher in the SH than SC group and the ET prevented this increase in the TH group. Resting bradycardia was observed only in the TC (296±20 bpm) group compared with SC (365±42 bpm). Triglycerides (211 %) and intrarenal lipid deposition (30%) increased in the SH group compared with SC, however the ET was able to prevent both responses in TH. Total cholesterol was not different among groups. In conclusion, our results showed that ET was efficient to prevent hypertension, dyslipidemia and the increase in intrarenal lipid deposition in rat fed a high‐fat diet since the weaning. Supported by Mackpesquisa.