bluuebunny/crossref_metadata_2025 huggingface dataset:
Euphol from Euphorbia tirucalli negatively modulates transforming growth factor‐beta (TGF‐β) responsiveness correlate with TGF‐β receptor segregation inside membrane rafts.
Authors: Ying‐Pin Chen , Chun‐Lin Chen
Published: June 2021

Transforming growth factor‐β (TGF‐β) responsiveness in cultured cells can be modulated by TGF‐β partitioning between lipid raft/caveolae‐ and clathrin‐mediated endocytosis pathways. Lipid rafts/caveolae is membrane microdomain with important roles in cell survival signaling. Cholesterol is important for the structure and function of these microdomains. Our previous studies shown that cholesterol, alone or complexed in lipoproteins, suppresses TGF‐β responsiveness by lipid‐raft segregation of TGF‐β receptors and facilitating rapid degradation of TGF‐β and thus suppressing TGF‐β‐induced signaling. Euphol is an euphane‐type triterpene alcohol, is structurally similar to cholesterol and has a wide range of pharmacological properties including anti‐inflammation and anti‐cancer. In the present study, we demonstrated that euphol suppress TGF‐β signaling by moving TGF‐β receptor from non‐lipid raft to lipid‐raft microdomains in plasma membrane and increased degradation of TGF‐β receptors.
Source Tracking of Fecal Contamination Along San Luis Obispo (SLO) Creek
Authors: Shapiro Dillon , Kent Jason , Zuleta Maria , Kitts Christopher , Black Michael , VanderKelen Jennifer
Published: June 2021

The primary goal of this project was to identify the biological sources of fecal contamination within and around the Mission tunnel of SLO Creek. Fecal coliform bacteria are used as an indication of fecal contamination that may include disease‐causing bacteria and viruses. Using a library‐based approach, we were able to track the possible sources of this contamination by analyzing specific regions of DNA within isolated E. coli samples. Environmental samples were collected from the Mission Tunnel mouth and underneath the Marsh Street Bridge and E. coli was isolated and identified using biochemical tests. Two regions of DNA (16‐23 and 23‐5 ITS) were amplified from each bacterial isolate and analyzed by a novel bacterial strain typing method known as pyroprinting. These pyroprints were matched to the Cal Poly Library of Pyroprints, which contains data for approximately 7000 E. coli isolates from various wildlife, human, domesticated animal, and environmental sources. Out of a total 843 isolates collected from SLO Creek, 25% matched isolates in the database. Approximately 72% of these matches were to unique hosts, with the majority of unique matches being to cow (51%) and pigeon (25%) isolates. Other matches include those to samples from humans (7%), horses (5%), turkeys (4%), and chickens (3%). The bias toward cow isolates (despite the absence of cows in the immediate area) and the low percentage of total matches indicates that more samples from different hosts need to be collected for the pyroprint library in order to gain a better picture of fecal contamination sources.
Functional Association Profiling of Metabolic Landscape and Microbiome in Chronic Pressure Ulcer Wounds
Authors: Ammons Mary Cloud , Copie Valerie
Published: June 2021

Chronic, non‐healing wounds contribute significantly to the suffering of patients with co‐morbidities in clinical population with mild to severely compromised immune systems. Normal wound healing proceeds through a well‐described process. However, in chronic wounds this process seems to become dysregulated at the transition between resolution of inflammation and re‐epithelialization. Bioburden in the form of colonizing bacteria is a major contributor to the development of chronic wounds such as pressure ulcers. However how the microbiome influences the wound metabolic landscape is unknown. Here, we have used a Systems Biology approach to determine the functional association between the taxonomic and metabolomic profile of wounds colonized by bacteria. Pressure ulcer biopsies were harvested from primary chronic wounds and sectioned into top and bottom sections prior to analysis of microbiome by pyrosequencing and analysis of metabolome using 1H nuclear magnetic resonance (NMR) spectroscopy. Bacterial taxonomy revealed that wounds were colonized predominantly by three main phyla, but differed significantly at the genus level. While taxonomic profiles demonstrated significant variability between wounds, metabolomic profiles shared significant similarity based on the depth of the wound biopsy. Functional association between taxonomy and metabolic landscape indicated significant wound‐to‐wound similarity in metabolite enrichment sets and metabolic pathway impacts, especially with regard to amino acid metabolism. To our knowledge, this is the first demonstration of a statistically robust correlation between bacterial colonization and metabolic landscape within the chronic wound environment.
A Potential Role of H. Pylori PPIase in the Transformation of Gastric Epithelial Cell and the Development of Gastric Cancer
Authors: Liu Jun
Published: June 2021

Helicobacter pylori (H. pylori) is the causative agent of chronic gastritis and gastric cancer, the second leading cause of cancer mortality worldwide. H. pylori colonizes the stomach in more than half of the world's population and has been classified as a class I carcinogen. H. pylori produces several virulence factors, including CagA, VacA and urease. Because the clinical outcomes of H. pylori infection do not necessarily correlate with the presence of these proteins, there is a need to identify new H. pylori factors that may modulate the clinical course of H. pylori‐associated diseases. Recently, we identified several gastric cancer‐associated H. pylori genes, including FKBP‐type peptidyl‐prolyl cis‐trans isomerase (PPIase‐FKBP). In this study, we show that co‐culture of gastric epithelial cells (AGS) with PPIase‐FKBP deletion H. pylori strain prevented H. pylori‐induced proliferation of AGS cells. Consistent with this observation, ectopic expression of H. pylori PPIase‐FKBP promoted AGS cell proliferation and anchorage‐independent growth. To gain insight into the biochemical mechanism of PPIase‐FKBP‐induced effect, early signal events involved in mitogenic signal pathways were evaluated. Expression of PPIase‐FKBP induced increases in basal as well as EGF‐stimulated phosphorylation of ERK and EGF receptor at Tyr1086. Pretreatment with MEK inhibitor completely blocked PPIase‐FKBP‐induced cell proliferation, suggesting that the action of PPIase‐FKBP is mediated by activation of ERK1/2 mitogenic signal pathway. H. pylori PPIase‐FKBP may represent a novel target for therapeutic management of gastric cancer patients.
Persister Cell Control Mechanisms in Uropathogenic Escherichia coli
Authors: Law William , Leatham‐Jensen Mary , Cohen Paul , Camberg Jodi
Published: June 2021

Persister cells are a subpopulation of bacteria that demonstrate high tolerance to antibiotics, but revert to sensitivity after antibiotics are removed. The mechanism for induction of the persister cell state and antibiotic tolerance is not completely understood but likely occurs through the establishment of dormancy. Some of the suggested mechanisms for persister cell formation in Escherichia coli include: toxin‐antitoxin systems, starvation, gene regulation by (p)ppGpp, and stochastic formation. In our study we examine the mechanisms behind persistence in the E. coli strain CFT073, a uropathogenic isolate, which forms elevated levels of persisters compared to the laboratory strain MG1655. However, CFT073 lacks many of the type II toxin‐antitoxin pairs associated with modulating persister cell formation. In addition, global stress response is impaired in the CFT073 isolate used in these studies, since it contains a five base‐pair insertion in the rpoS gene, the stationary phase sigma factor and regulator of the global stress response. This insertion results in the expression of truncated RpoS. We compared several CFT073 strains mutated at the rpoS locus for the ability to form persister cells, as well as a strain deleted for lon, a protease important for modulating toxin‐antitoxin function. To identify additional regulators of persister cell formation in CFT073, we performed minitransposon mutagenesis to isolate mutant strains, and screened mutants for persister cell formation. As many pathogenic bacterial species, including CFT073, cause recalcitrant infections attributable to persister cell activity, these studies will identify additional mechanisms underlying the development of bacterial persistence in these organisms.
Novel Resistance Mutations to ALK Tyrosine Kinase Inhibition Arising During Cell‐Culture Drug Selections in T‐Cell Lymphoma
Authors: Groysman Matthew , Dipak Amin Amit , Tapia Edgar , Rajan Soumya , Schatz Jonathan
Published: June 2021

Anaplastic Large Cell Lymphoma (ALCL) is a rare T‐cell lymphoma. The Anaplastic Lymphoma Kinase (ALK)‐positive molecular subtype of ALCL is driven by constitutive activation of the ALK tyrosine‐kinase domain, typically through chromosomal translocation. The most common rearrangement, t(2;5)(p23;q35), results in the NPM‐ALK fusion kinase. Recently, two ALK‐specific tyrosine kinase inhibitors (TKIs), crizotinib and ceritinib, have been approved for clinical use in ALK‐positive lung cancer while another inhibitor, alectinib, is in early‐stage clinical trials. We developed TKI‐resistant ALK+ ALCL cell line subclones from Karpas‐299, SUP‐M2 and SU‐DHL‐1. Each line was cultured in increasing concentrations of crizotinib or ceritinib until viable at 500nM and 150nM, respectively. We identified three novel mutations in our resistant lines affecting the ALK kinase domain: R1192P, I1171S, and F1174L. We cloned wild‐type and mutant NPM‐ALK into a murine retroviral vector and infected an IL3‐dependent murine pro‐B cell line FL5‐12. The infected lines were withdrawn from IL‐3, leading to their IL3‐independent, oncogene‐dependent transformation. We evaluated the half‐maximal inhibitory concentration (IC50) of crizotinib, ceritinib and alectinib for wild‐type ALK‐dependent and mutant ALK dependent FL5‐12 cells, through luminescence based viability assays (Table 1). We found that all three mutations confer resistance to both crizotinib and ceritinib to varying degrees. I1171S was the only mutation to confer resistance to alectinib, while R1192P and F1174L remain sensitive. Our findings have significant clinical implications in matching specific TKIs to patient disease based on ALK mutational status.

NPM‐ALK Genotypes
Average Crizotinib IC50 (nM)
Crizotinib Standard Error of the Mean
Average Ceretinib IC50 (nM)
Ceretinib Standard Error of the Mean
Average Alectinib IC50 (nM)
Alectinib Standard Error of the Mean

Wild Type
171.85
7.19
34.03
2.00
2.84
.33

R1192P
366.8
17.56
97.19
5.40
5.47
.29

I1171S
479.825
28.9
57.7
1.24
43.185
4.34

F1174L
245.05
15.9
57.015
3.62
3.87
.68
Landscape Phage Probes for Prostate Specific Antigen
Authors: Fagbohun Olusegun , Petrenko Valery , Abraham Eisenstark
Published: June 2021

Prostate‐specific antigen (PSA) is a cardinal marker for molecular diagnosis and proper management of prostate tumors. Therefore, development of specific probes for this marker is paramount for precise molecular diagnosis and targeted delivery of therapeutics to prostate cancer cells. Screening of the multibillion landscape libraries proffers a simple and economic approach for isolating target‐specific phage probes. Landscape phage libraries are collections of filamentous phages displaying random guest peptides on all 4,000 surface domains of the major coat protein. They serve as a rich source of bioselective materials that can be used in different areas of technology and medicine, including gene delivery, drug delivery and imaging of tumor cells. Novel highly specific and selective phage probes against prostate specific antigens were selected from 8‐mer library using immobilized PSA as targets. The selected clones were identified by nucleotide sequencing of their DNA and translation into proteins using software. Eight phage probes were isolated and tested for their selective interactions with PSA in comparison with control targets, two of the phage probes selectively interacted with PSA. The selected phage probes can serve as promising ligands for development of imaging and therapeutic devices precisely targeted to prostate tumors.
New Strategies for Probing and Inhibiting CDK4/cyclin D Hyperactivity in Melanoma
Authors: Prével Camille , Pellerano Morgan , Bihel Frederic , Meunier Laurent , Morris May
Published: June 2021

Uncontrolled proliferation of melanocytes results in melanoma, one of the most aggressive forms of skin cancer, which further metastasizes into a deadly form of cancer. The p16‐cyclin D‐CDK4/6‐retinoblastoma protein pathway is dysregulated in 90% of melanomas and activation of this pathway potently cooperates with mutant BRAF or NRAS in transformation of melanocytes. CDK4/cyclin D kinase hyperactivation, associated with mutation of CDK4, amplification of cyclin D or loss of p16INK4a leads to increased risk of developing melanoma. This kinase therefore constitutes a key biomarker and an emerging pharmacological target.

The limited efficacy of current therapeutics for treatment of melanoma and the lack of diagnostic assays to detect the early onset of this disease call for new generations of targeted therapeutics and novel approaches to probe melanoma‐specific biomarkers. To this aim, we engineered a fluorescent biosensor which reports on CDK4 kinase hyperactivity in vitro and in living cells, which we employed to develop a diagnostic assay using human skin biopsies. We further designed biosensors for drug discovery purposes, which discriminate compounds that target the interface between CDK4 and cyclin D, or that interfere with conformational dynamics of CDK4 activation loop, which were successfully applied to screen for novel and potent classes of CDK4 inhibitors

Our work highlights the potency of fluorescent biosensors for developing diagnostic strategies to monitor cancer‐associated alterations in CDK4 activity as well as drug discovery programmes.
Bovine lactoferrin is a transcription factor stimulating IL‐1 beta gene expression in the small intestine
Authors: Jiang Rulan , Lönnerdal Bo
Published: June 2021

Lactoferrin (Lf) is an iron‐binding glycoprotein present in high concentration in human milk. It is a pleiotropic protein and involved in diverse bioactivities. Since Lf is partly resistant to proteolysis in the gastrointestinal tract, Lf may play important roles in intestinal development. Human Lf (hLf) is internalized by human intestinal epithelial cells (Caco‐2) and translocates to the nucleus where it functions as a transcription factor to up‐regulate expression of genes, such as IL‐1 beta. Bovine Lf (bLf) shares 69% amino acid identity with hLf and has been shown to have similar physiological functions as hLf. Therefore, we hypothesized that bLf is able to enter the nucleus of Caco‐2 and transactivate gene transcription. Confocal microscopy and immunoblotting using nuclear fractions showed that bLf was internalized and localized to the nucleus of Caco‐2. After Caco‐2 cells were treated with bLf, IL‐1 beta transcription was two fold up‐regulated as revealed by RT‐PCR. To determine if bLf binds to the promoter of IL‐1 gene, an IL‐1 beta promoter fragment containing a hLf potential DNA binding site were then inserted to a luciferase promoter‐less vector and luciferase assays were conducted. The results show that bLf binds to the IL‐1 beta promoter and activates IL‐1 beta transcription. Results from chromatin immunoprecipitation (ChIp) assays also demonstrated that bLf interacts with the potential hLf DNA binding site in the promoter of the IL‐1 beta gene in vivo. In conclusion, bLf is able to enter the nucleus of human intestinal epithelial cells, bind to the promoter of IL‐1 gene and functionally regulate gene transcription. It is therefore possible that bLf may insert some of the biological activities of Lf in human milk. Supported by Mead Johnson Nutrition.
SUGAR CONSUMPTION IN THE UNITED STATES‐ FOOD GROUP CONTRIBUTIONS BY EATING OCCASIONS: RESULTS FROM NHANES 2011‐12
Authors: Albertson Ann , Holschuh Norton
Published: June 2021

Data from NHANES 2011‐12 were analyzed to determine the food group contributions by eating occasion to total sugar consumption for the total population and by age groups. Total sugar consumption currently accounts for approximately 23% of daily energy for Americans ages 2+ years. Soft drinks, non‐flavored milk and milk products, desserts, candy/sweets and fruit are the leading contributors of total daily sugar for the U.S. population (15%, 12%, 10%, 8%, and 8%, respectively). Twenty‐one percent of total daily sugar is consumed at breakfast. Non‐flavored milk and milk products, candy/sweets, fruit juices, cereals and other grain products are the leading contributors at breakfast (17%, 13%, 11%, 10%, and 10%, respectively). Eighteen percent of total daily sugar is consumed at lunch with leading contributors including soft drinks, grain products (excluding cereals and desserts), fruit drinks, non‐flavored milk and milk products and fruit (20%, 11%, 8%, 8% and 8%, respectively). Twenty‐three percent of total daily sugar is consumed at the dinner occasion and the leading sources are similar to lunch; soft drinks, grain products (excluding cereals and desserts), non‐flavored milk and milk products, fruit and fruit drinks (20%, 12%, 11%,9 % and 8%, respectively). Thirty percent of total daily sugar is consumed at snacking occasions. Leading sources include dessert foods, non‐flavored milk and milk products, candy/sweets, soft drinks, and fruit juices (16%, 14%, 14%, 13%, and 12%, respectively). The food group contributions at the four eating occasions vary slightly by the three age groups examined (2‐12, 13‐18 and 19+ years).
DETERMINANTS OF HIGH PLASMA TRIGLYCERIDE LEVELS: A BRAZILIAN COMMUNITY‐BASED STUDY
Authors: Kano Hugo , Manda Rodrigo , Moreto Fernando , Burini Franz , Burini Roberto
Published: June 2021

The modern lifestyle, represented by food inadequacy and sedentary behavior is probable the major etiological factor for the burden of contemporary chronic diseases. It is well established that abnormal blood lipids levels appear as a key factor in inflammation and atherogenesis and the management of the altered lipids are directly related to clinical outcomes. This study aimed to analyze the major determinants associated with higher blood triglycerides levels in free‐living adults. In a cross‐sectional study, 582 individuals (80.4 % female), 54.6 + 10.8 years old, were clinically assessed for admission into a lifestyle‐changing program (“Move For Health”). Initially, they were assessed for medical anamneses, blood pressure, physical fitness, body composition/anthropometric (Body Mass Index, Waist Circumference, Body Fat), biochemical (Total cholesterol, HDL‐c, LDL‐c, Gama‐GT, CRP, Triglycerides (TG), Glucose) and cardiorespiratory fitness. Food intake was analyzed by 24h recall questionnaire. TG levels were divided in quartiles for comparison with all co‐variables. ANOVA one way was performed to investigate the difference between triglyceride quartiles followed by Tukey's post hoc. The determinants of TG levels were determinated by Multiple Regression Backward Stepwise (p<0.05). Higher TG levels were associated with higher WC, BMI, glucose, gama gt, CRP, and lower VO2max, weekly physical fitness and HDL‐c. The multiple regression showed that the levels of gama gt (β:0.34) and CRP (β: 0.48) were responsible for 51.2% of the high TG levels. Thus, higher plasma TG levels are determined by either obesity and abdominal high adiposity and their consequences, lower aerobic capacity, inflammation and hyperglycemia‐induced stress.
Genetic Variants in GLUT14 Gene Enhance Susceptibility to Inflammatory Bowel Disease
Authors: amir shaghaghi mandana , Bernstein Charles , Eck peter
Published: June 2021

Oxidative stress is implicated in the initiation and progression of inflammatory bowel disease (IBD). Dehydroascorbic acid (DHA) is actively imported into cells via glucose transporters (GLUTs) and contributes to the vitamin C antioxidant cycle. This study aimed to determine if the recently indentified GLUT 14 gene is associated with susceptibility to IBD and the clinical phenotype. Participants in the population‐based Manitoba IBD Cohort Study were phenotyped using the Montreal classification. Genomic DNA samples from persons with Crohn's disease (CD, n=162) and ulcerative colitis (UC, n=149) and ethnically matched controls (n=142) were genotyped for eight GLUT 14 polymorphisms, using TaqMan assays. Logistic regression was used to determine the case‐control association of genotypes for each polymorphism with CD, UC, and overall IBD risks. Genotype‐phenotype associations were tested by multinomial logistic regression, using SAS software 9.2. Variant rs2889504‐A was associated with increased susceptibility to UC, CD, and overall IBD (OR: 3.60, 95% CI: 1.95‐6.64; OR: 4.68, 95% CI: 2.78, 8.50; OR: 4.14; 95% CI: 2.37‐7.22 respectively). The rs10846086‐G allele was associated with an approximated 3‐fold increased risk of UC, CD, and IBD. Variant rs12815313‐T was associated with increased susceptibility to CD and overall IBD OR: 2.12, 95% CI: 1.33‐3.36; OR: 1.85, 95% CI 2.41 (1.24‐2.77; respectively). These findings provide a novel opportunity toward individualized nutritional therapy for patients carrying the disease‐associated genotype. Further studies are now needed to elucidate the pathogenetic mechanism. (Supported by Peter Eck's Canada Research Chair)
Immunomodulation of innate immune cells by Sutherlandia frutescens: A transcriptomic analyses
Authors: Lei W. , Lu Y. , Hou J. , Li J. , Browning J. , Eichen P. , Cheng J. , Lubahn D. , Folk W. , Sun G. , Fritsche K.
Published: June 2021

Sutherlandia frutescens (SF) is a plant found in southern portions of Africa with a rich history of traditional use as a beverage to ameliorate some of the symptoms associated with cancer and HIV‐AIDS. To date no research has reported the impact of SF on global gene regulation. Therefore, we designed an experiment to evaluate the effects of SF treatment on gene expression in a murine macrophage cell line (RAW 264.7). Cells were exposed to SF extract or a vehicle control for 1 hr prior to activation with LPS and IFNγ. Gene expression at 8 & 24 hr post‐stimulation were analyzed using RNA sequencing. Immune‐stimulation caused ~2600 genes to be up‐regulated and another 2600 genes to be down‐regulated at 8 hr and at 24 hr post‐stimulation up‐ and down‐regulated genes expanded to ~4100 and >4500, respectively. SF treatment resulted in 547 differentially expressed genes (DEGs) at 8 hr post‐stimulation and for most of these genes (~64%) SF altered the direction of their expression. While the total number of genes modulated at 24 hr post‐stimulation was much greater than at 8 hr, the portion that were differentially regulated by SF treatment was lower (1369 out of 3625, ~38%). Pathway analysis identified that many of these DEGs participate in inflammatory signaling pathways, such as NF‐kB and MAPK. These findings were consistent with previously reported anti‐inflammatory activity for SF and should help us better understand the immune‐modulating properties of this botanical.
The Effects of Lying, Sitting and Standing on Energy Expenditure in Young, Adult Women
Authors: Popp Collin , Bridges William , Najm Nadia , Traylor Daniel , Jesch Elliot
Published: June 2021

Background

Sedentary behavior and the increased risk for metabolic disease are well established. Standing at work or at home to decrease time sedentary can increase daily energy expenditure, however, little research has focused on the energy expenditure differences between various sedentary positions (i.e., lying, sitting).

Objective

The purpose of this study was to investigate the energy expenditure of lying, sitting and standing.

Methods

Twenty‐eight young, healthy females reported to the metabolic laboratory where each completed all positions (lying, sitting and standing) in random order. Inspired and expired gases were collected for 45‐minutes (15 minutes for each position), and all three positions were measured during one continuous test. Oxygen consumption (VO2) and carbon dioxide production (VCO2) were measured to estimate resting energy expenditure (REE) and respiratory quotient (RQ). Statistical analyses used repeat measures ANOVA to analyze all variables and post hoc t‐tests.

Results

There were positional differences for most variables. Standing VO2 (L/min) was 9.5% and 7.7% greater than lying VO2 (L/min) and sitting VO2 (L/min), respectively. Standing RQ was lower than lying RQ, but not different from sitting RQ. Standing REE (kcal/min) was 9.0 % greater than lying REE (kcal/min), and 7.1% greater than sitting REE (kcal/min), however, lying REE and sitting REE were not different from each other. Relative standing REE (kcal/min/kg) was 8.8% greater than lying REE (kcal/min/kg) and 6.4% greater than sitting REE (kcal/min/kg).

Conclusion

Our results indicate that standing has small, but significant effect on energy expenditure when compared to lying and sitting.
Validation of Photography for Determining Food Intakes of High School Students
Authors: Barrett Sheila , Kyle Ashley , Skaradzinski Veronica , Miu Leong Calvin
Published: June 2021

Purpose

To determine if photography is a reliable data collection tool for measuring the food intakes of high school students who participate in the National School Lunch Program (NSLP). Secondly, to compare the energy and macronutrient content of the NSLP meals with meals eaten when not at school.

Method

Researchers visited one high school in Illinois and measured the amounts of food consumed by the students using photographs of before and after food trays and weighed food wastes for comparisons. Three researchers not involved with the weighing of the food wastes, estimated the percentage wastes from the photographs. Estimated percentage wastes were converted into gram amounts by using the actual gram weight of the food items served. Actual wastes were weighed and compared with the estimated gram wastes. Students completed a demographic questionnaire and a three day food record which was used to determine the contribution of calories and macronutrients consumed when not at school. Twenty‐two students from grades 9‐12 participated in this pilot study, mean age was 16.41±1.30.

Results High inter‐rater reliability was found for estimated and actual gram amounts of food consumed (r= 0.957, p<0.001). Paired samples t‐test of calories and macronutrients indicated significant differences (p<0.05) for meals eaten when not at school with those provided by the NSLP lunch meal. Overall, the NSLP meals provided fewer calories and macronutrients.

Conclusion

Photography can be used to determine the food consumption of the NSLP participants in future studies. Contrary to earlier studies, the NSLP meals provided fewer calories and macronutrients than did the meals eaten when not at school. Future studies using a larger sample size are needed.
Weight Loss with Liquid Formula Diets in Obese Patients with and without Diabetes
Authors: Leslie Wilma , Taylor Roy , Lean Michael
Published: June 2021

Aim

To provide a data synthesis, using systematic methods, of published results comparing weight losses following Very Low Calorie (<800kcal/day VLCD) or Low Energy Liquid‐formula (>800kcal/day LELD) diets in patients with type 2 diabetes and non‐diabetic patients.

Methods

Medline (1946‐2013) search to identify published studies of VLDC/LELDs. All studies in T2D patients were included. Limits for studies in non‐diabetic patients were: VLCD/LELD >8weeks duration, sample size >50 and within last 7 years. Mean weekly rate of weight loss during VLCD/LELD were weighted for study number.

Results

Weighted rates of weight loss in the 16 studies included were as follows: 6 in T2DM patients alone, (n=130, mean BMI 33.6‐40kg/m2) ‐1.4kg/week; non‐diabetic obese (n=1031, mean BMI 30‐48 kg/m2) ‐1.3kg/week, and 4 studies that included both patient groups (n=116, mean BMI 36.6‐42.6 kg/m2) ‐1.4kg/week (diabetic) and ‐1.4kg/week (non‐diabetic).

Conclusions

These data show that weight loss with LELD is very similar for obese patients with and without T2DM and support the use of LELD to achieve new weight loss/ maintenance targets of >15‐20% for those with severe and medically complicated obesity.
Influence of High Fat Diet on Macrophage Behavior and Metabolic Processes in Obesity Sensitive and Resistant Mice: Role of Monocyte Chemoattractant Protein 1
Authors: Cranford Taryn , Enos Reilly , Velazquez Kandy , McClellan Jamie , Nagarkatti Mitzi , Murphy E
Published: June 2021

PURPOSE

Adipose tissue macrophages are thought to be a major player in the dysregulation of metabolic and inflammatory processes associated with high fat diet (HFD)‐induced obesity. Monocyte chemoattractant protein 1 (MCP‐1) is a chemokine that recruits macrophages to adipose tissue (AT) and has been implicated in manifestations associated with obesity. The purpose of this study was to examine the role of MCP‐1 in response to HFD feedings. We used both C57Bl/6 (obesity sensitive) and FVB/N (obesity resistant) mice in order to determine obesity versus HFD effects. A HFD that was designed by our laboratory and is similar to a standard American diet was used given its clinical relevance.

METHODS

C57Bl/6 and FVB/N wild‐type (WT) and MCP1‐/‐ mice at 4 wks of age were fed either a HFD or a low fat diet (LFD) for 16 wks. Body composition was analyzed monthly and HOMA index was determined at mice sacrifice at 20 wks of age. AT mRNA expression of chemokines, macrophage markers and protein expression of inflammatory markers were measured using qPCR and western blot, respectively. Macrophage infiltration into AT were examined using immunohistochemistry and flow cytometry.

RESULTS

As expected, HFD induced obesity and increased insulin resistance (IR) comparably in the obesity sensitive C57Bl/6 WT and MCP1‐/‐ mice. Surprisingly, the HFD MCP1‐/‐ in the supposed diet resistant FVB/N strain exhibited increased obesity and IR whereas the HFD WT mice were not different from LFD mice. AT macrophage accumulation increased in HFD WT groups for both strains, regardless of obesity status. Interestingly, the response in HFD MCP1‐/‐ was dependent on the strain.
Acute Ingestive Effects of Agave Nectar Versus Sucrose in Healthy Young Adults
Authors: Cravinho Amanda , Hammon Megan , Rieger Kelsey , Kern Mark
Published: June 2021

Less refined sweeteners may impact metabolism differently from their more refined counterparts. We have demonstrated that agave nectar may beneficially alter metabolism in mice in comparison to sucrose. Since agave nectar is particularly rich in fructose, we hypothesized that it would promote lower acute glycemic responses but may promote gastrointestinal (GI) discomfort in comparison to sucrose. We also assessed satiety responses to the sweeteners. Healthy young adults (n=21) between the ages of 20‐45 years completed two randomly ordered feeding trials in which they consumed 200 ml of a test beverage providing 50 grams of carbohydrate from sucrose or raw agave nectar mixed in water. Glycemic responses were examined by finger prick at baseline and 15, 30, 45, 60, 90 and 120 minutes post‐feeding. Symptoms of GI discomfort (nausea, cramping, distention and flatulence) were self‐reported using visual analogue scales at the same timepoints. Satiety was assessed by visual analogue scale at baseline and every 15 minutes for two hours. Agave nectar consumption elicited significantly (p<0.05) lower blood glucose concentrations over time compared to sucrose. Although the magnitude of nausea was relatively low for both trials, greater (p<0.05) nausea was reported during the agave nectar trial. No other symptom of GI discomfort was differentially affected. No significant differences were observed for satiety responses between the two sweeteners. Results suggest that raw agave nectar as a less refined sweetener promoted lower blood glucose responses in young healthy adults as well as modest GI discomfort. Future research should assess the acute and chronic ingestive effects of this sweetener in healthy individuals and those with metabolic disorders.
Dietary Glucogenesis by Mucosal α‐Glucosidase is Influenced by the Internal Molecular Structure of Starch Hydrolysates
Authors: Muniandy Anbuhkani , Lin Amy
Published: June 2021

Four small intestinal α‐glucosidases, N‐ and C‐terminal subunits of maltase‐glucoamylase (MGAM) and sucrase‐isomaltase (SI), releases glucose from starch molecules at the final stage of digestion in the small intestine. In our previous study, we hypothesized that the digestibility at α‐glucosidase level is determined by the internal molecular structure of starch hydrolysates. We later reported that the branch pattern of starch internal molecules influences the digestion. The objective of this study is to further test the hypothesis that the branch amount and the distance between branches influence the rate and extent of glucose release from α‐amylase hydrolysate. Two starches [waxy (wx) maize and potato] with different internal structure were hydrolyzed by pancreatic α‐amylase. The hydrolysate consists of linear glucans and branched dextrin (α‐limit dextrin,α‐LD), and they were separated by size‐exclusion chromatography. α‐LD was digested by Ct‐MGAM in vitro, and the rate and extent of glucose release were examined. Both wx maize and potato hydrolysates consist of 30‐35% of α‐LD, and the molecular size of their α‐LD was DP (average degree of polymerization)13 and 17, respectively. The amount of released glucose from wx maize and potato α‐LD was 30 and 40 mg (p<0.05), and the rate of glucose release as represented by the catalytic efficiency was 0.13 and 0.28, respectively. In conclusion, α‐LD with longer distance between branches and loose branched structure (potato in this study) generated higher glucose amount at higher rate by Ct‐MGAM. In the future, starches with different type of internal structure needs to be further examined. This project was sponsored by Univ. of Idaho and Whistler Center for Carbohydrate Research at Purdue Univ.
A randomized control trial of sugar‐sweetened and artificially sweetened beverages and intrahepatic fat in overweight subjects
Authors: Campos V , Despland C , Schneiter Ph , Brandejsky V , Kreis R , Boesch Ch , Tappy L
Published: June 2021

We conducted a 12‐week trial involving 28 overweight male and female adults (BMI >25) consuming 2 or more servings/day sugar‐sweetened beverages (SSB). Participants were randomized to a 12‐week intervention replacing SSB with artificially sweetened beverages (ASB, 8M, 7F) or to a control arm without intervention (C, 6M, 7F). Intrahepatocellular lipid concentration (IHCL, 1H‐magnetic resonance spectroscopy), visceral adipose tissue volume (VAT, magnetic resonance imaging) and fasting concentrations of selected metabolic parameters were measured before and after intervention.

Consumption of SSB was similar in both groups at inclusion. IHCL was reduced to 74 ±10.7% of initial values in the ASB (p<0.01) but did not change in C. ASB however did not significantly change body weight (‐1.4±1.6 vs +0.8±0.6 kg in C), VAT (‐98.4 ±78.4 vs ‐11.8 ±73.4 cc in C) or fasting glucose, insulin, uric acid, ASAT, ALAT, TG, cholesterol or HDL‐cholesterol concentrations.

15 participants (8 in the ASB group, 7 in the C group) had hepatic steatosis as defined by IHCL > 60 mmol/L. They had significantly higher body weight, VAT, TG, uric acid, ASAT, ALAT, and lower HDL‐cholesterol than 13 participants without hepatic steatosis. In this subgroup, ASB was associated with significant (p<0.05) decreases in IHCL (‐43.0±12.6%), ALAT (‐19.7±7.8%) and ASAT (‐9.9±4.0 %).

These results indicate that, in high SSB consumers 1) body weight and visceral fat volume are major determinants of hepatic steatosis, 2) replacing SSB with ASB reduces intrahepatic fat, most likely due to decreased total energy intake.

This work was supported by the Swiss National Science Foundation and the Raymond Berger Foundation.
Effect of Korean Red Ginseng water extract on the hepatic fatty acid and cholesterol synthesis in HepG2 cells
Authors: Lee Mak‐Soon , Moon Sohee , Lee Seohyun , Kim Yangha
Published: June 2021

Korean Ginseng, the root Panax ginseng CA Meyer, has been widely used as a traditional herbal medicine in East Asian countries. In this study, we investigated the effects of Korean Red Ginseng water extract (KRGE) on hepatic lipid accumulation in HepG2 cells. First, we investigated the cytotoxic effects of KRGE on HepG2 cells. Cytotoxicity was unaffected in the concentrations of 1–50 μg/ml of KRGE.Intracellular lipid accumulation was assessed by the quantification of intracellular triglyceride and cholesterol contents. KRGE decreased hepatic triglyceride and cholesterol levels. Moreover, KRGE suppressed expression of fatty acid synthase (FAS) and 3‐hydroxy‐3‐methyl glutaryl coenzyme A (HMG‐CoA) reductase, a rate‐limiting enzyme in hepatic cholesterol synthesis. These results suggest that KRGE may reduce hepatic lipid accumulation by inhibition of FAS and HMG‐CoA reductase expression in HepG2 cells. Consequently, KRGE may be beneficial as a functional food ingredient to improve various hepatic diseases by reducing hepatic lipid accumulation.
Potato Phenolics Modulate Rate of Glucose Transport in a Caco‐2 Human Intestinal Cell Model
Authors: Moser Sydney , Aragon Ingrid , Furrer Amber , Van Klinken Jan‐Willem , Kaczmarczyk Melissa , Lee Byung‐Hoo , Hamaker Bruce , Ferruzzi Mario
Published: June 2021

Potatoes are a rich source of phenolic compounds. While there is evidence for beneficial effects of some phenolics to modulate carbohydrate digestion and glycemic response, the effects of potato phenolics on these responses is not known. The objectives of this study were to characterize the phenolic profile of fresh potatoes (purple, red, or white; 2 varieties each) and finished potato chips and to examine the potential for potato phenolic extracts (PPE) to modulate starch digestion and intestinal glucose transport. We hypothesized that PPEs would inhibit in vitro α‐amylase and α‐glucosidase activity and decrease rate of d7‐glucose transport in a Caco‐2 human intestinal cell model. PPEs were found to be rich in anthocyanins (98, 11 and 0 mg/100 g dw) and chlorogenic acids (519, 425 and 157 mg/100 g dw) for purple, red and white varieties respectively. While no significant effects were observed on starch digestion, PPEs (from whole potato and peel) were found to significantly (p<0.05) decrease the rate of glucose transport over a 60 min period by 4.50‐83.89% (3.0µmoles/min for d7‐glucose control relative to 0.48‐2.21 µmoles/min for PPEs). Furthermore, PPEs at 100 μM were found to decrease 60 min transport of d7‐glucose by 11.81‐83.64% relative to d7‐glucose controls. These results suggest that potato phenolics merit further consideration as bioactive agents that may affect rate of intestinal glucose transport. Funding provided by Pepsico Global R&D.
Plasma alkylresorcinols as a biomarker for whole grain intake and predictor of metabolic syndrome risk
Authors: MacArthur Michael , Magnanti Steven , Landberg Rikard , Lofgren Ingrid
Published: June 2021

Alkylresocinols (AR) are dietary phenolic lipids obtained mainly from wheat and rye bran. Research shows that plasma AR homologues are potentially suitable biomarkers for whole grains (WG) intake. The objective of this ancillary cross‐sectional study was to investigate relationships between WG intake, plasma AR and cardiometabolic risk factors in college students (18‐24 y). Subjects (n=130) completed 3 24‐hr dietary recalls and had fasting blood drawn on 2 nonconsecutive days. Plasma was analyzed for AR concentration (homologues C17:0‐C:25:0) by gas chromatography‐mass spectrometry; total cholesterol, triglycerides, HDL‐C and glucose by enzymatic assay; and LDL‐C by the Friedewald equation. There was a correlation between subjects' total WG intake and plasma AR (ρ=0.246, p=0.006). Regression analysis showed that total WG intake explained about 6% of the variance in plasma AR concentrations (r2=0.058, p=0.01). Although triglycerides was the only cardiometabolic risk factor associated with plasma AR (ρ=0.27 p=0.003), logistic regression showed that plasma AR was a significant predictor of subjects displaying 2 or more metabolic syndrome criteria (OR=1.009, 95% CI=1.003‐1.015, p=0.004). These results show a weak linear relationship between self‐reported WG intake and plasma AR in persons consuming few WGs, and suggest that plasma AR may be a predictor of metabolic syndrome risk in college students.
A Natural Herbal Beverage Exhibits Significant Cytoprotection and Promotes Nrf‐2 Activation in Cells
Authors: Gumpricht Eric , Kumar Rahul , Hussain Anowar , Sabarwal Akash , Ramteke Anand , Cho Suk , Deep Gagan
Published: June 2021

Oxidative stress is a central feature of chronic diseases including cancer, atherosclerosis, diabetes, the aging process, and exposure to environmental toxins. Therefore, activating cytoprotective defense mechanisms through natural, non‐toxic nutritional interventions might provide an important avenue for improving human health. In the present study, we evaluated the effects of an herbal‐rich beverage (Cleanse For Life [CFL]) in both homogeneous solutions and in cellular models of oxidative stress and toxicity using HepG2 and human prostate PWR‐1E cells. In solution, CFL scavenged hydroxyl (IC50: 50.4 μg/ml), ABTS+(IC50: 124.7 μg/ml), and DPPH (IC50: 256 μg/ml) free radicals while demonstrating metal‐chelating activity toward ferric ion (IC50: 60.5 μg/ml). In HepG2 cells, CFL (10‐100 μg/ml) dose‐dependently inhibited H2O2‐induced oxidative stress (21‐67%, p蠄0.001). CFL (10‐25 μg/ml) also decreased expression of cytochrome P450s E1 (up to 30%) and A1/A2 (up to 46%) but significantly increased expression of glutathione transferases mu and pi. Moreover, CFL strongly induced (1.7‐fold) expression of Nrf‐2 (Nuclear factor erythroid‐derived 2‐like 2), a primary transcription factor of antioxidant enzymes. Finally, exposing HepG2 or PWR‐1E cells to the environmental toxin polychlorinated biphenyl (PCB‐153) promoted substantial cytotoxicity which was abrogated by CFL. Together, these results suggest that the herbal bioactives present in CFL activate cellular antioxidant defenses and confer strong cytoprotection in cells. Future studies should investigate effects of CFL in modulating oxidative stress and detoxification in humans.
Polyphenols Extracted From Grape Powder Induce Lipogenesis and Glucose Uptake During Differentiation of Murine 3T3‐F442A Preadipocytes
Authors: Torabi Sheida , DiMarco Nancy
Published: June 2021

Assessing the effect of dietary ingredients on lipogenesis and glucose uptake in adipocytes may clarify the risk/benefit of recommending to individuals with obesity and insulin resistance. Grape powder extracted polyphenols (GPEP) modulate inflammation in human adipocytes. We hypothesized that GPEP increase intracellular fat accumulation and glucose uptake during differentiation of 3T3‐F442A preadipocytes. Total polyphenols were extracted and measured based on gallic acid equivalents (GAE). There were 2167 mg of GAE of polyphenols in 100 g of grape powder (GP). 3T3‐F442A cells were incubated with GPEP from 125 – 500 µg GP/mL of media until day 8 of differentiation when the cells were collected for different assays. Adipo‐Red assay and Oil Red O staining showed that GPEP induced, in a dose‐dependent manner, an increase in intracellular triglyceride content of adipocytes. Concomitantly, GPEP increased, in a dose‐dependent manner, glucose uptake by 3T3‐F442A cells. No changes in cell viability were measured by Trypan blue staining, suggesting that these effects were independent of cytotoxicity. Western‐blot showed that GPEP upregulated total and membrane protein level of GLUT4 in 3T3‐F442A adipocytes. Ly294002 (10 µmol/L), a phosphatidyl‐inositol 3 kinase (PI3K) inhibitor, reversed the effects of GPEP on cellular lipid content. Real‐time qPCR showed that GPEP increased mRNA expression of fatty acid synthase, lipoprotein lipase, and adiponectin, while it decreased mRNA expression of leptin and insig‐1. Our results indicate that GPEP may induce adipocyte differentiation via upregulation of GLUT4, PI3K and adipogenic genes.
Molecular Mechanism of FXa Inhibition by Cufaxapin, the Main Salivary Anticoagulant of the West Nile Virus Vector Culex quinquesfaciatus
Authors: Calvo Eric , Chagas Andrezza , Ramirez Jose , Ribeiro Jose
Published: June 2021

The saliva of mosquitoes contains complex mixtures of bioactive compounds inhibiting hemostasis in their vertebrate host. Here we have identified and characterized the main salivary anticoagulant (Cufaxapin) from Culex quinquesfaciatus. Cufaxapin, an atypical member of the serpin superfamily, is a tight, non‐covalent, competitive inhibitor of FXa with a calculated affinity (KD) of 56pM as determined by SPR analysis. Cufaxapin inhibits prothrombinase activity and significantly increases both PT and aPTT. Isothermal titration calorimetry showed that Cufaxapin binds to FXa in a stoichiometric (1:1) way with a favorable ΔH of ‐34.12±0.26 kcal/mol. This interaction does not undergo detectable conformational change as determined by CD spectroscopy. The absence of binding to factor X or catalytic site blocked FXa indicates that Cufaxapin specifically targets the catalytic site of FXa. Site‐directed mutagenesis in the amino acid sequence and length of Cufaxapin RCL do not cause any significant alteration in the molecular stability or structural flexibility of the inhibitor. Cufaxapin also prevents laser‐induced carotid thrombus formation in vivo and showed a potent antiinflammatory effect on carrageenan and FXa‐induced paw edema formation. Proinflammatory cytokines IL6, IL8 and MCP1 were significantly reduced in cultured human endothelial cells treated with FXa amixed with Cufaxapin (1:2) when compared to cells treated with FXa (40nM) or Cufaxapin alone (80nM). Silencing of Cufaxapin via dsRNAi exhibited a significant reduction in its mRNA accumulation and protein levels in SG. However no effect in feeding success was observed in silenced mosquitoes. The role of Cufaxapin for Culex mosquitoes blood feeding and survival is discussed.
Modeling Steatosis and Lipid‐Mediated Cell Injury Using Induced Pluripotent Stem Cell‐Derived‐Hepatocytes
Authors: Ignatius Irudayam Joseph , Contreras Deisy , French Samuel , Arumugaswami Vaithilingaraja
Published: June 2021

Non‐alcoholic fatty liver disease (NAFLD) is characterized by steatosis with varying degrees of hepatocellular injury, fibrosis and cirrhosis. The iPSC‐derived hepatocytes (iHep) model is a novel system to study the molecular and cellular basis of fat‐mediated injury. The objective of this study is to investigate invitro steatosis and lipotoxicity of free fatty acids in iPSC‐derived hepatocytes. Methods: A three stage differentiation protocol was used for derivation of metabolically active hepatocytes. The iHeps were exposed to various doses of free fatty acids (FFA), palmitic acid (PA) and oleic acid (OA) for 24 hours. Cell viability, apoptosis, oxidative stress, lipid accumulation, glucose consumption and apoB release were studied. Results: The FFA treatment showed a dose‐dependent accumulation of fat in the cytoplasm of iHeps. Micro and macro‐vesicular patterns of lipid accumulation were detected. A significant increase in apoB release was observed in the FFA group. Interestingly, cells loaded with lipid had increased consumption of glucose. At higher concentrations (over PA 200um and OA400um), the lipid induced apoptosis by activation of caspase 7. The mitochondrial membrane potential was higher at 24 hrs in the treatment group compared to that of control indicating perturbation in mitochondrial respiration. In conclusion, iPSC‐hepatocytes can be used for modeling fatty liver disease.