bluuebunny/crossref_metadata_2025 huggingface dataset:
Pathogen‐induced lipid coalescence in the host membrane causes downstream activation of Rho GTPase (99.3)
Authors: Krachler Anne‐Marie
Published: June 2021

We recently discovered a family of bacterial adhesins (Multivalent Adhesion Molecules, MAMs) which mediate initial contact between pathogen and host cells during infection. MAMs are composed of tandem arrays of six to seven mammalian cell entry (mce) domains, each recognizing a phosphatidic acid lipid ligand in the host cell membrane. We utilized bacteriomimetic tools to reconstruct the process of pathogen adhesion to host cells independent of other bacterial components and virulence factors. This revealed that MAMs, through binding to phosphatidic acid ligands with high affinity, induce lipid coalescence in the host cell membrane. Clustering of phosphatidic acid ligands triggers downstream activation of the small GTPase RhoA and actin rearrangements. In vivo infection studies with Vibrio parahaemolyticus demonstrated that MAM‐triggered RhoA activation facilitates breaching of a polarized epithelial cell layer by the pathogen. This is an unprecedented mechanism utilized by a bacterial pathogen to manipulate host cells and facilitate infection.

Grant Funding Source: This work was supported by an EMBO fellowship (ALTF 938‐2011).
Biochemical and Microbiological Studies of the Semen, Mouth swab and Serum of infertile Nigeria Male (LB232)
Authors: Samuel Aderonke , Olagunju O
Published: June 2021

There is an increase in male infertility in Nigeria mostly due to abnormal seminal fluid which may be found in up to 60% of infertile male, resulting in a multi‐ factorial health related issues.

This research evaluates the relationship between oral health and infertility in Nigeria men.

A total of 43 men were recruited, grouped into 30 infertile men and 13 controls; aged 30 to 50 years, bio data and health related information were collected. Microbial spectrum of seminal and mouth swab was analysed, the following biochemical parameters: lipid profile, testosterone, prolactin, LH and FSH were analysed using serum samples.

The average semen volume, concentration and motility of infertile group was significantly lower than the normal group, with a corresponding increase in abnormal spermatozoa morphology of the test group compared to the control (p<0.05).

The spectra of bacteria revealed Staphylococcus aureus as the most common bacteria isolate in both the semen culture and the mouth swab of infertile candidates. The level of Testosterone was significantly decreased in the infertile men (Test) group, while the levels of Prolactin, FSH and LH were significantly increased. HDL showed a significant decrease in the test group compared to the control; however there is no significant difference in Creatinine and Urea level of both groups.

The results did not confirm any mouth infection that has a direct correlation to infertility status of men; however, there is an increase in seminal fluid infections with decreasing sperm concentration, volume, motility and morphology. Hence, there is no direct association between mouth infection and male infertility.
HIF and Notch non canonical pathways affect the emergence and differentiation of Cardiovascular Progenitor Cells (LB270)
Authors: Calderon Alexander , Ramírez‐Bergeron Diana
Published: June 2021

Adaptive responses to low Oxygen (O2) tension (hypoxia) are integral to embryogenesis, tumorigenesis, and tissue ischemia. Developing embryos begin to experience a hypoxic microenvironment promoting the development of the cardiovascular system. Early differentiation events focus the generation of Flk‐1, Brachyury double positive cardiovascular progenitor cells, hemangioblasts, which have hematopoietic, endothelial, and cardiomyocyte potential. Our model system utilizes embryonic stem (ES) cells, which can be cultured and differentiated under physiologic conditions (3 % O2) into embryoid bodies (EBs) mimicking early development Hypoxia Inducible Factor (HIF) is a major transcriptional player required for the temporal differentiation of these progenitor cells. We propose that HIF also plays a significant noncanonical role in mediating the generation and differentiation of hemangioblasts cells by interacting with the Notch signaling pathway. Using chemically and genetically modified mouse ES cells, differentiation experiments are being developed to explore the connection between these two pathways. Our present data demonstrate that hypoxia influences the expression of Notch receptors (Notch‐1 and ‐4), ligands (Delta‐4) and downstream Notch targets (Hrt‐1 and ‐2).

Grant Funding Source: Supported by: R25‐HL03152 NHLBI Summer Research Program at CWRU
Fasciola hepatica fatty acid binding protein inhibits the activation of TLR4 (LB288)
Authors: Ortiz Ivelisse , Cabán Kimberly , Espino Ana
Published: June 2021

Lipopolysaccharide (LPS) is a major component of the outer membrane of Gram‐negative bacteria and is a potent activator of the innate immune system. The immune response to LPS can lead to septic shock syndrome if the inflammatory response is amplified and uncontrolled which can be fatal. The receptors that recognize LPS are lipid binding protein (LBP), CD14, MD12 and Toll‐like Receptor 4 (TLR‐4) which all cooperates in the pathway activation. Today, there is much interest to develop TLR antagonists to control the innate immunity and regulation of innate immune responses. TLR antagonists currently under development are structural analogs of agonists and probably bind to the receptor but fail to signal or pathway activation. However, there is still a need for effective molecules that could prevent or circumvent the negative effect of inflammatory responses during a variety of human diseases.

We demonstrated that the protein Fasciola hepatica fatty acid binding protein (FABP) has antagonist properties through TLR‐4 after LPS stimulation. FABP binds to human CD14 co‐receptor of HEK 293 cells to block the activation of NF‐kB induced by LPS, functioning thus as antagonist of TLR4. We also demonstrated that FABP inhibited the production of inflammatory molecules as TNF‐α, IL‐12A and NOS production by human monocytes derived macrophage (MDM) in vitro. This results directed to explore the potential of FABP as a new class of drug against septic shock bacterial or others inflammatory diseases in which TLR4 are involved.
Biodegradation of venlafaxine (LB289)
Authors: Claybourne James , Shi Edmund , Strand Stephanie , Schultz Melissa , Snider Mark
Published: June 2021

The antidepressant venlafaxine (VEN) is being detected in surface waters in high concentrations relative to other detected pharmaceuticals. The high concentrations of VEN in surface waters are the result of the inefficiency of current organic compound removal techniques employed by wastewater treatment facilities (WWTF). Current research involves understanding the pathways by which bioremediation of compounds such as VEN can be achieved. Previous data suggests that VEN is rapidly degraded in microcosms (t1/2 ~ 2 days) when cultured with aerobic sludge from WWTPs. The present study investigates the aerobic bacteria and the environment of the activated sludge that are contributing to the biodegradation pathway of VEN. Liquid media cultures containing VEN (100 ppm) and trace metals (TMs) were inoculated with (1) bacteria previously isolated from aerobic sludge with the capacity for growing on minimal media in which venlafaxine was the sole carbon source and (2) variety of known pseudomonas species, and examined for the ability to degrade VEN. Aliquots from the cultures were taken with time, the VEN was extracted and quantified by liquid chromatography tandem mass spectrometry. Preliminary evidence showed VEN was degraded only in the presence of TMs. The emergence and subsequent disappearance of VEN metabolites of in cultures containing TMs provides additional evidence for the bacterial degradation of VEN. Identifying the bacteria will allow for the subsequent determination of the degradation pathway and the genes and mechanisms associated with its degradation.

Grant Funding Source: Supported By Research Corporation
Dietary intake and nutritional status of children and adolescents with HIV‐infection (LB295)
Authors: Chaud Daniela , Furtado Miriam , Machado Alisson , Abreu Edeli
Published: June 2021

Introduction: HIV‐infection is more severe in children and adolescents due to their growth and development.

Objective: To evaluate the dietary intake and nutritional status of children and adolescents with HIV‐infection.

Methods: This cross‐sectional study was performed with 12 children and adolescents with HIV‐infection living in na institution located in São Paulo, Brazil. The variables studied were dietary intake, by qualitative analysis of the daily menu offered by the institution, and nutritional status, obtained by the measurement of height and weight to subsequent classification according to the World Health Organization growth curves.

Results: 50% of subjects were male and 50% were female. The menu offered by institution had low offer of fruits and high offer of sugars. Regarding nutritional staus, 83.3% of participants were categorized as eutrophic.

Conclusion: The menu offered, despite the great offer of sugar, was adequate with respect to most of the food groups. The most children and adolescents showed eutrophic.
Comparative effectiveness of popular diet programs on changes in body composition and visceral adipose tissue in women (LB297)
Authors: Jung Y , Lockard B , Baetge C , Levers K , Galvan E , Jagim A , Simbo S , Byrd M , Oliver J , Koozehchian M , Dalton R , Khanna D , Sanchez B , Kresta J , Horrell K , Leopold T , Cho M , Springer S , Rivera A , Cerda C , Chang C , Rasmussen C , Kreider R
Published: June 2021

The purpose of this study was to determine if different types of exercise and diet interventions promote greater changes in visceral adipose tissue (VAT). 127 sedentary women (46±12 yr, 45.5±5% body fat, 35.1±5 kg/m2) were randomized to participate in a control group (C) or the Curves Complete® program with online support (CC), Weight Watchers® Points Plus (WW), Jenny Craig® (JC), or Nutrisystem® Advance Select™ (NS) weight loss programs for 12‐wks. DEXA body composition with VAT determination was obtained at 0 & 12 wks. MANOVA revealed significant (p=<0.001) time and group x time (p=<0.001) effects. Participants in the CC group experienced greater loss in fat mass (C ‐0.0±2.1; CC ‐4.6±3.7; WW ‐2.2±2.7; JC ‐3.5±3.3; NS ‐2.1±2.7 kg, p<0.001), less loss in FFM (C 0.2±2.3; CC ‐0.3±2.7; WW ‐1.7±2.4; JC ‐1.8±2.3; NS ‐1.8±2.4 kg, p=0.01), and greater reductions in percent body fat (C ‐0.0±1.7; CC ‐3.1±2.5; WW ‐0.7±2.6; JC ‐1.4±2.5; NS ‐0.5±1.7 %, p<<0.001). VAT mass, volume and area decreased over time (p=0.004) with no significant differences among groups. Changes in VAT mass significantly correlated with changes in fat mas (r=0.20) and body fat (r=0.22). Results indicate that different types of diets can differentially affect changes in body composition but promote proportional changes in VAT.

Grant Funding Source: Supported by Curves International (Waco, TX)
Analysis of the validity of a carbohydrate intolerance questionnaire III (LB303)
Authors: Koozehchian M , Levers K , Galvan E , Coletta A , Dalton R , Jung Y , O’Connor A , Goodenough C , Simbo S , Seesselberg C , Bonin B , Sanchez B , Barringer N , Rasmussen C , Greenwood M , Kreider R
Published: June 2021

The purpose of this study was to examine the relationship of responses to a carbohydrate intolerance (CI) questionnaire to body composition and markers of health. 108 women (31.6±13 yrs, 34.7±7% body fat, BMI 25.3±4) donated fasting blood samples, had body composition and health measures determined, and underwent a 75g, 2‐h OGTT. Being less than 25 lbs overweight correlated with age (r=0.19) and DBP (r=0.27). Getting hungry at meals correlated with BMI (r=0.23), antisocial behavior (r=0.37) and having a poor appetite/skipping meals (r=0.39). Having few food cravings correlated with waist circumference (r=‐0.20), mental confusion (r=0.23), being 25 lbs overweight (r=‐0.27), having temporary food cravings (r=‐0.23), and must have foods (r=‐0.30). Consistent eating habits correlated with RHR (r=‐0.22), FFM (r=‐0.19), being overweight throughout life (r=‐0.20), and skipping meals (r=‐0.20). Eating 3 times/d correlated with OGTT glucose AUC (r=‐0.91), antisocial behavior (r=‐0.19), being overemotional (r=‐0.21), leg cramps/blurred vision (r=‐0.27), drowsiness (r=‐0.19), and skipping meals (r=‐0.43). Maintaining weight gain correlated with waist (r=0.21) and hip (r=0.20) circumference, fat mass (r=0.21), body fat (r=0.22), headaches (r=0.20), mental confusion (r=0.28), leg cramps/blurred vision (r=0.28), and yawning (r=0.26). Results revealed that CI questions moderately correlated with body composition, health markers, and/or questions related to CI.

Grant Funding Source: Curves International, Inc. (Waco, TX)
Prevalence of functional constipation and its risk factors among university students (LB328)
Authors: Chaud Daniela , Olivon Edilene , Machado Alisson , Abreu Edeli
Published: June 2021

Introduction: Functional constipation is a complex and multifactorial disease; its symptoms affects the quality of life by compromising bowel functions and is a risk factor for the development of cancer.

Objective: To determine the prevalence of functional constipation and its risk factors among university students. Methods: This cross‐sectional study was performed with 52 university students from a private university located in São Paulo, Brazil. A self‐administered questionnaire about demographic characteristics, lifestyle and bowel habit was applied. For the assessment of presence of functional constipation was used Rome III criteria. The prevalence of this symptom was assessed by chi‐square test, using the SPSS software, version 20.0.

Results: The mean age of the sample in study was 22.7±3.7 years old, with 46.2% male and 53.8% female. The prevalence of functional constipation was 30.8% and was statistically higher for the variable liquid intake (p<0,05). No difference was observed in their distribution according gender, age, use of tobacco, number of meals, consumption of foods rich in dietary fiber, presence of constipation in the family and physical activity practice.

Conclusion: The prevalence of functional constipation found in this present study can be considered high. There was difference in the prevalence of this symptom only to the variable liquid intake.
Polymorphisms in 4 exon, 3 intron, 6 intron of p53 gene, 31 codon of p21 gene and in 11 exon of apc gene in the dna of uranium ore workers in republic of kazakhstan (LB33)
Authors: Balmukhanov Timur , Miroshnick Tatiyana , Kazymbet Polat , Belkozhaev Ayaz , Bakhtin Meirat , Botbaev Dauren
Published: June 2021

Genotypes distribution in 4 exon (G/C, Arg/Met) in codon72, 3 intron (16 bp duplication), 6 intron (G/A) of p53 gene, 31 codon of p21 gene and in 11 exon (T/C) of APC gene in DNA of 203 uranium mine workers were compared with polymorphisms in DNA of 255 controls in two ethnic groups in Republic of Kazakhstan. It were evaluated the statistically significant differences (p<0.05) in the genotypes distribution and allele frequencies in 3 intron of p53 gene in Kazakh ethnic group Chi‐square for genotypes was equal to 5,631, p = 0,018, for alleles: Chi‐sq was equal to 5,866, p = 0,015. Russian ethnic group in genotypes distribution and allele frequencies, for genotypes value of Chi‐sq was equal to 7,167, p=0,027, for alleles: Chi‐sq=5,567, p=0,018. In 31 codon of p21 gene also in Kazakh ethnic group the statistically significant differences were observed in genotypes distribution Chi‐sq =7,703, p=0,006 and in alleles frequency: Chi‐sq =4,453, p=0,035. In 11 exon of APC gene the statistically significant differences in genotypes distribution and in alleles frequency between miners and controls were not found in both groups.

The presented data is a result of preliminary investigation indicating to the necessity of enlarging of the size of cases and controls in cohorts by measuring of the same indexes in other uranium mines of Kazakhstan and in uranium enriching plant.
Inhibitory effects of gossypin on RANKL‐induced osteoclast differentiation and bone resorption in murine macrophages (LB364)
Authors: Park Seon Ju , Lee Eun‐Jung , Kim Yun‐Ho , Shin Ji‐Eun , Kang Young‐Hee
Published: June 2021

Bone‐remodeling imbalance between bone formation by osteoblasts and bone resorption by osteoclasts results in the metabolic bone diseases such as osteoporosis. Gossypin is a naturally‐occurring bioflavonoid seen in vegetables and fruit, as being a treatment for melanoma. The present study investigated that 1‐20 μM gossypin suppressed RANKL‐induced bone resorption in Raw 264.7 macrophages. To differentiate to osteoclasts, macrophages were treated with 50 ng/ml RANKL for 5 days. RANKL enhanced multi‐nucleated osteoclast formation and tartrate‐resistance acid phosphatase activity, which was dose‐dependently attenuated by gossypin. Western blot analysis revealed that gossypin suppressed RANKL induction of integrin β3 and carbonic anhydrase II in macrophage‐derived osteoclasts. These results demonstrate that gossypin retarded the differentiation and activation of osteoclasts to be integrated on bone tissue. Therefore, gossypin is a potent agent in alleviating bone resorption of osteoclasts.

Grant Funding Source: Supported by National Research Foundation of Korea (2012012946) and Ministry for Food, Agriculture, Forestry and Fisheries
Association between sugar‐sweetened beverage intake and proxies of acculturation among Hispanic and non‐Hispanic white adults (LB373)
Authors: Park Sohyun , Blanck Heidi , Dooyema Carrie , Ayala Guadalupe
Published: June 2021

Because there is limited information on the association between acculturation of Hispanics and their sugar‐sweetened beverage (SSB) intake, we examine this question in the 2010 National Health Interview Survey, a nationally representative sample of US adults. Analyses were restricted to Hispanic and US‐born non‐Hispanic (NH) white (n=17,142) respondents. The outcome was daily SSB intake (regular soda, fruit drinks, sports drinks, energy drinks, and sweetened coffee/tea drinks). Exposure variables were race/ethnicity and proxies of acculturation (language of interview, birthplace, and years living in the US). We used multivariable logistic regression to estimate odds ratios (OR) for exposure variables associated with SSB intake 蠅1 time/day, adjusting for sociodemographics, region, and weight status. The odds of drinking SSB 蠅1 time/day was significantly higher among Hispanics who completed the interview in Spanish (OR=1.65 vs. US‐born NH whites) and foreign‐born Hispanic adults who lived in the US for 5–<10 years (OR=2.72), those who lived in the US for 10–<15 years (OR=2.90), and those who lived in the US for 蠅15 years (OR=2.41) than those who lived in the US for <5 years. But, birthplace was not associated with SSB intake. The acculturation process is complex and these findings contribute to identifying important subpopulations that may benefit from targeted intervention approaches to reduce SSB intake.
Mechanisms of β‐carotene transfer from placenta to embryo in mammals (LB406)
Authors: Costabile Brianna , Kim Youn‐Kyung , Rodas Rebeka , Wassef Lesley , Shete Varsha , Spiegler Elizabeth , Zuccaro Michael , Quadro Loredana
Published: June 2021

Vitamin A is an essential nutrient for mammalian embryonic development. The most abundant precursor of vitamin A in the human diet is β‐carotene (BC). The enzyme β‐carotene 15‐15’‐monooxygenase (CMOI) cleaves BC to form retinaldehyde which is oxidized to form retinoic acid, the biologically active form of vitamin A. BC can be cleaved by β‐carotene 9’‐10’‐monooxygenase (CMOII) to form apocarotenoids which can function as retinoid precursors.

The mammalian embryo obtains BC from the maternal circulation and converts it to retinaldehyde via CMOI. However, BC transfer across the maternal‐fetal barrier is not fully understood. We demonstrated that embryos from wild‐type dams on high vitamin A diet (220 IU/g) and treated with BC at 13.5 dpc, had lower BC levels compared to embryos from wild‐type dams on vitamin A sufficient diet (14 IU/g). Regulatory mechanisms involving regulation of placental BC secretion may therefore prevent embryonic retinoid toxicity. Ex vivo placenta culture and in vivo placenta injection experiments showed that placental BC secretion rate is attenuated in wild‐type dams on high vitamin A diet. It is unknown how BC is transferred from placenta into fetal circulation. We hypothesized that transfer occurs via lipoproteins, which transport BC in the bloodstream. ApoB‐containing lipoprotein formation was reported in placenta that also expresses microsomal triglyceride transfer protein (MTTP), a lipoprotein assembly and secretion regulator. We showed that placental MTTP transcription levels were elevated in dams on the sufficient diet and supplemented with BC. ApoB mRNA and protein levels were also increased in this group. BC may control its own secretion from the placenta through lipoprotein secretion depending on maternal vitamin A status.

Grant Funding Source: Supported by NIH
Peanut butter increases kale β‐carotene absorption and conversion to vitamin A in pre‐school children (LB417)
Authors: Muzhingi Tawanda , Tang Guangwen , Yeum Kyung‐Jin , Bermudez Odilia , Siwela Andrew
Published: June 2021

Brassica oleracea var. acephala (kale) is a rich source of β‐carotene that can be used to provide vitamin A to vitamin A deficient preschool children in Africa. This study was designed to determine the effect of peanut butter on the vitamin A equivalence and bioconversion efficiency of kale β‐carotene to vitamin A in preschool children. Deuterium labeled kale was grown in a hydroponic medium with 23 atom% 2H2O during grain development. The kale β‐carotene showed the highest abundance of enrichment as [2H9] β‐carotene. Preschool children (n = 37; age 12‐36 mo) were randomly assigned to consume 50g cooked kale (1.5mg β‐carotene content) with either peanut butter (PBG) or with lard (LG). On day 1 after a baseline blood draw; all subjects consumed the labeled kale doses and 1 mg [13C10] retinyl acetate capsule. Blood samples were collected from five subjects per‐time point per group over 21 d. Serum samples processed were analyzed by using NCI‐GC/MS for the enrichments of labeled [2H]retinol from kale from [2H9]β‐carotene and from [13C10]retinol from reference dose. The area under the curve (AUCs) of molar enrichment at days 1, 2, 3, 6, 15, and 21 after the labeled doses was 21.1 and 27.9 (nmole) for [2H] retinol from LG and PBG kale [2H9] β‐carotene respectively. The AUC of [13C10] retinol from was 223.4 (LG) and 314.45 (nmol) (PBG) respectively. The calculated conversion factors were 19.0 and 19.3 to 1 by weight for LG and PBG respectively. Peanut butter had higher β‐carotene absorption response than the lard group. This study showed that kale β‐carotene is efficiently absorbed and converted to vitamin A in the presence of fat in children. Therefore, kale should be promoted as a complementary food for children in developing countries who are vulnerable to VAD and general malnutrition.

Grant Funding Source: IAEA
A clinical evaluation to determine the safety, pharmacokinetics and pharmacodynamics of an inositol‐stabilized arginine silicate dietary supplement in healthy adult males. (LB418)
Authors: Kalman Douglas , Feldman Samantha , Samson Adam , Krieger Diane
Published: June 2021

The purpose of this study was to characterize the PK/PD of Nitrosigine™, an inositol‐stabilized arginine silicate dietary supplement. Ten healthy males, 26.7 ± 5.4 years, took Nitrosigine™ qd for 14 days and attended test visits on Days 1 and 14. Fasting blood and saliva collections were performed pre‐dose and at 0.5, 1, 1.5, 2, 3, 4, 5 and 6 hours post‐dose for plasma arginine, serum silicon and salivary nitric oxide + nitrite (NO). After Day 1 dose, arginine levels increased (p<0.01) at 0.5, 1, 1.5, 2, 3 and 5 hours and silicon levels increased (p<0.05) at 1 and 1.5 hours. Day 1 PK parameters for arginine were: CMax 27.25±4.86 µg/ml and tMax 1.05±0.50 and for silicon: CMax 2.77±0.74 µg/ml and tMax 2.15±1.75. After Day 14 dose, arginine levels increased (p<0.05) at 0.5, 1, and 1.5 hours, and silicon levels increased (p<0.01) at 1, 1.5, 2, and 3 hours. After 14 days of use, baseline arginine trended towards being higher than baseline Day 1 (p=0.0645), and 4‐hour post‐dose plasma arginine was significantly higher (p=0.0488). Although not a significant difference, NO increased in four subjects and stayed the same in six subjects at 0.5 hours after the first dose (p=0.125). After 14 days of use, baseline NO levels increased in six subjects and stayed the same in four subjects (p=0.031). No safety concerns were raised by this study.
Palmitoleic acid enhances glucose uptake and glycerol‐3‐phosphate generation in adipocytes (LB426)
Authors: Bolsoni‐Lopes Andresss , Festuccia William , Faria Talita , Chimin Patricia , Torres‐Leal Leonardo , Curi Rui , Lima Fabio , Alonso Vale Maria Isabel
Published: June 2021

Palmitoleic acid (16:1n7) enhances whole body glucose disposal, suppresses hepatic steatosis and modulates triacylglycerol (TAG) metabolism in adipocytes. Here, we investigated the effects of 16:1n7 on glucose uptake and TAG synthesis by primary adipocytes. METHODS: Primary epididimal adipocytes from C57BL6 mice treated with 16:1n7 (300 mg/kg/day), oleic acid (18:1n9, 300 mg/kg/day) or water by gavage for 10 days were evaluated for 2‐deoxy‐D‐[3H]‐glucose uptake on basal and insulin‐stimulated (10 nmol/liter) conditions, phosphorylated and total AKT content, 14C‐glucose incorporation into fatty acids or glycerol and 14C‐acetate incorporation into fatty acids. RESULTS: Treatment with 16:1n7 or 18:1n9 did not affect body weight, food intake, masses of white adipose depots and plasma levels of free fatty acids, insulin and glucose. Treatment with 16:1n7 increased basal (~3 fold) and insulin‐stimulated (~2 fold, p<0.05) glucose uptake on epididimal adipocytes compared with water and 18:1n9 treatment. There were no changes on acetate or glucose incorporation into fatty acids among treatments, but the 16:1n7 increased glucose incorporation in glycerol‐TAG (H2O=227.2±9.1; 18:1n9=226.6±24.6; 16:1n7=307.2±19 nmol/106 cells, p<0.05). The treatments did not change the phosphorylated and total AKT content. CONCLUSION: 16:1n7 raised basal and insulin‐stimulated glucose uptake without affecting AKT. Furthermore, 16:1n7 increased the generation of glycerol‐3‐phosphate from glucose on epididimal adipocytes without affecting fatty acid synthesis from this hexose.

Grant Funding Source: Supported by FAPESP
Differential Risk Perceptions toward Illness guide development of a prevention of stunting intervention in rural Mozambique (LB477)
Authors: Kodish Stephen , Aburto Nancy , Dibari Filippo , Gittelsohn Joel
Published: June 2021

Objective: To inform the behavior change communication (BCC) component of a stunting prevention program in Cabo Delgado, Mozambique, the World Food Programme (WFP) supported the government’s Scaling Up Nutrition (SUN) movement to understand how community members perceive the risk of young children’s illnesses associated with poor nutritional status.

Methods: Among community leaders and caretakers of children under 2 years in Macua and Mwani cultural groups, data were collected in a three‐phase, iterative process from May ‐ August 2013 using free listing (n = 80), pile sorting (n = 67), and in‐depth interviewing (n = 34). Free list illness terms were analyzed for statistical salience and pile sort data were analyzed using multi‐dimensional scaling (MDS). Interview data were coded inductively using a Grounded Theory approach.

Results: Biomedical nutrition illness terms are largely absent from the cultural domains of the groups studied. The most important illnesses to the Macua group include malaria, stomachache, and headache, whereas the most salient illnesses to the Mwani group are cough, stomachache, and fever. Community members do not associate the causes of these illnesses with food and nutrition, but do categorize their signs and symptoms in relation to perceived severity. Stunted growth, wasted or emaciated appearance, and slowed cognitive development are perceived to be less severe than other disease signs and symptoms.

Conclusion: Behavior change models posit that a person’s perceived susceptibility and severity of a health problem are important for his or her behavior change. Because our results indicated that among these cultural groups, illnesses associated with poor nutritional status were not considered important threats to a child’s health, WFP supported the government to develop a BCC strategy that appeals to other core cultural values of the ethnic groups rather than solely focusing on the program’s potential impact upon nutritional status. Widespread sensitization efforts were also made around chronic malnutrition prior to intervention roll out.
Analysis of breast cancer markers and phosphoproteins in clinical tumor tissues using rapid two‐temperature formalin fixation (LB483)
Authors: Lerch Melissa , Theiss Abbey , Chafin David , Otter Michael , Baird Geoffrey
Published: June 2021

Breast cancer can be categorized into molecular subtypes that respond to specific therapies based on the expression of three protein biomarkers: estrogen receptor (ER), progesterone receptor (PR), and receptor tyrosine‐protein kinase erbB‐2 (HER2). Using a novel rapid two‐temperature formalin fixation method, we analyzed a set of clinical breast carcinoma samples and compared our results with classifications generated by standard processing (i.e. a clinical diagnosis generated by the institution’s clinical laboratory and pathologists). To ensure objectivity in staining assessment, we scored immunohistochemical staining by automated image analysis, and demonstrated that the quantity of IHC staining observed with our rapid fixation protocol is highly correlated with the score determined by the clinical pathologist on each case. Three additional phoshphoproteins were analyzed by IHC, protein kinase B (pAKT), p38 mitogen‐activated protein kinase (MAPK), and proline‐rich AKT substrate of 40kDa (pPRAS), and the rapid fixation protocol was found to preserve IHC signals from all three biomarkers.
TGFB‐induced protein ig‐h3 is essential for the growth of human colorectal carcinoma liver metastases (LB490)
Authors: Blomme Arnaud , Palacios Ana , Delvaux David , Delvenne Philippe , Detry Olivier , Castronovo Vincent , Turtoi Andrei
Published: June 2021

Transforming growth factor‐beta‐induced protein ig‐h3 (TGFBI) is an important actor of the extracellular matrix. While elevated expression of TGFBI has been reported in human cancer, its role still remains controversial. We show here for the first time that TGFBI is a relevant factor for VEGF expression in cancer cells.

As evidenced by immunohistochemistry, high level of TGFBI is a constitutive feature of human colorectal cancer liver metastases (CRC‐LM). To assess its function in cancer, we silenced TGFBI in human CRC cells. The suppression of the TGFBI protein decreased cancer cells migration and proliferation. In vivo, TGFBI silencing resulted in a 50% decrease in the volume of CRC tumors grown on a chicken chorioallantoic membrane (CAM). Low vascularization of these tumors suggested an additional anti‐angiogenic effect. Indeed, we found that TGFBI knockdown in SW1222 cells altered the secretion of VEGFA and led to an impairment of the subsequent signaling pathway. This observation was in line with previous data showing that TGFβ was able to induce TGFBI in cancer cells, forging a direct functional link to angiogenesis.

Finally, to test if TGFBI could be a relevant target for antibody‐drug delivery, we injected a fluorescently labeled anti‐TGFBI antibody in mice bearing liver metastases. The in vivo data demonstrated that TGFBI is an accessible tumor target.

Taken together, our study shows that TGFBI is essential for the development of CRC‐LM and therefore represents a promising target for novel therapeutic approaches.

Grant Funding Source: Supported by FNRS and the University of Liege
PTEN‐induced kinase 1 (PINK1) and Protein Kinase A protects against mitochondrial dysfunction and dendritic loss in a toxin model of PD (LB500)
Authors: Das Banerjee Tania , Dagda Raul , Dagda Ruben
Published: June 2021

Decreased mitochondrial and cytosolic signaling governed by serine/threonine (ser/thr) kinases, such as PINK1 and PKA, underlie the etiology of Parkinson’s disease (PD) as evidenced in PINK1 deficient neurons, toxin models of PD, and in postmortem PD brain tissue. Given that PINK1 upregulates downstream PKA signaling to remodel dendrites and enhance neuronal survival, we hypothesize that PINK1 activates distinct subcellular pools of PKA to confer neuroprotection in a chemical model of PD. Here, we demonstrate that in primary neurons, pharmacological activation of PKA signaling, overexpression of PINK1, untargeted PKA or mitochondrial PKA but not a PKA binding deficient mutant of A‐kinase anchoring protein 1 (D‐AKAP1), significantly prevents the loss of dendrites induced by 6‐hydroxydopamine (6‐OHDA). Overexpression of untargeted and mitochondrial PKA increases the basal oxygen consumption rates of SH‐SY5Y cells while only untargeted PKA protects against 6‐OHDA‐mediated decrease in mitochondrial respiration as measured by a XFE24 Analyzer. Mechanistically, PKA signaling is amplified by PINK1 by elevating the total levels of PKA monomers and enhancing the levels of membrane bound PKA. Overall, we show that PINK1 and PKA form a unique signaling axis that confers neuroprotection in the 6‐OHDA model of PD.

Grant Funding Source: Supported by NIH grant P20GM103554
PAR4 Mediates an Elevated Risk for Thrombosis in Blacks Relative to Whites (LB602)
Authors: Tourdot Benjamin , Niisuke Katrin , Holinstat Michael
Published: June 2021

Coronary heart disease (CHD) is the leading cause of mortality in the US amongst all races. CHD disproportionality burdens blacks relative to whites and even after adjusting for other demographic parameters, blacks have a two‐fold increase in incidence of CHD and a decrease in long‐term survival. Occlusive thrombus formation is the underlying cause of mortality associated with CHD. Due to the importance of platelets in CHD and formation of occlusive thrombi, the response of platelets from blacks and whites to thrombin and its protease‐activated receptors (PAR1 and PAR4) was characterized. Platelets isolated from blacks and whites differentially responded to PAR4 but not PAR1. Specifically, platelets from blacks were hyperaggregable to low and intermediate doses of PAR4‐activating peptide (PAR4‐AP) compared to platelets from whites. The mechanism responsible for the racial difference in PAR4 mediated platelet reactivity however is still unknown. We hypothesize that changes in expression or activity of proteins known to participate in the PAR4 pathway are responsible for the racial difference in platelet PAR4 reactivity. To test this hypothesis, the PAR4 pathway in platelets from blacks and whites was characterized. No measurable difference was observed in the surface expression of PAR4 on platelets from blacks or whites however following stimulation with low dose PAR4‐AP (50uM), platelets from blacks had an increase in PAR4 signaling components compared to whites including intracellular Ca2+ release, RAP1 activation, and integrin αIIbβ3 activation. This data suggest that the kinetics of platelet activation through PAR4‐AP is hyper‐responsive in blacks compared to whites due to a shift in threshold downstream of the receptor. As PAR1 antagonists are currently in clinical trials and PAR4 antagonists are in pre‐clinical development, this work has important clinical implication as to which patients may benefit the most from selective PAR inhibition and is the first to suggest a racial component in the prevention of occlusive thrombus formation and stroke.

Grant Funding Source: RO1 MD007880
Characterization of cocaine‐ and amphetamine‐regulated transcript signaling in Rat2 cells (LB617)
Authors: Prinster Steven , Kanda Shilpy , Smith Rachel
Published: June 2021

The cocaine‐ and amphetamine‐regulated transcript (CART) was first isolated from rat striatum following the acute administration of cocaine or amphetamine. It was quickly determined that CART or CART peptide was expressed in a variety of tissues affecting a wide range of important processes. The function of these areas have grown to include feeding behavior, pleasure and reward, diabetes, ovarian follicle development, bone remodeling and anxiety. In spite of the relevance of these conditions as therapeutic targets, development of small molecules to modulate these systems has been impeded by the lack of an identified receptor for CART peptide. Localization studies suggest that CART and CART peptide are located in similar areas: hypothalamus, pituitary and adrenal gland. Cells isolated from these tissues have provided information delineating much of the current state of knowledge regarding the CART peptide receptor. We have characterized the signaling pathways activated by CART peptide in the Rat 2 fibroblast cell line. We observed a pertussis‐toxin sensitive inhibition of cAMP production, and increased phosphorylation of extracellular signal regulated kinase 1 and 2. These results suggest that the CART peptide receptor is more widely expressed than previously appreciated. These CART peptide studies in Rat 2 cells provide important information that will aid in the identification of CART peptide receptor.
B‐success supplement (abnigerian herbal remedy): Reproductive implications on on male albino rats (LB631)
Authors: Agbasi Patrick , Obi Ejatuluchukwu
Published: June 2021

We investigated the effects of B‐Success herbal supplement ( A Nigerian Herbal Remedy ) on the accessory sex organs of male Albino Rats. Twenty eight weight matched male Albino Rats were given 0.00, 315, 630, 945 mg/kg of B‐success herbal agent for a period of 90 days. The Rats had free access to deionized water and were fed with rat pellets for the entire period of study. The feed and fluid consumption of the animals were measured on daily basis while the body weight was measured weekly. Animals were anaesthetized after 90 days, bled and sacrificed. Epididymis, seminal vesicle and prostate were excised and weighed. The result showed significant (P蠄 0.05) decrease in both absolute and relative weights of seminal vesicle, prostate and epididymis in all the treated animals when compared with the control. The epididymal acid and alkaline phosphatase contents and semen count decreased significantly (p蠄 0.05) in all the treated animals when compared with the control. The DNA and protein contents of seminal vesicle (SV), prostate (P) and epididymis (E) of albino rats treated with the B‐success herbal supplement decreased significantly (p蠄 0.05) in all the treated animals when compared with the control. B‐success herbal supplement may have toxic effect on the accessory sex organs of male albino rats Key words: B‐success herbal supplement, seminal vesicle (SV), prostate (P) and epididymis (E), male albino rats.
Ageing differentially affects myogenic tone of coronary microcirculatory arteries according to vessel diameter (LB644)
Authors: Mansour Fathi , Hassan Mohammed , Albarwani Sulayma
Published: June 2021

Aging is the major cause of decline in cardiovascular function and for structural and functional alterations of coronary microvasculature. There is close association between cardiac function and coronary blood flow; hence, age‐associated impairment of cardiac function may be related to reduced coronary blood flow capacity. As myogenic tone has a major role in regulation of local blood flow, the aim of this study was to investigate changes in myogenic tone with aging and its heterogeneity based on resistance arteries size. Coronary resistance arteries from young (3 ‐ 4 months) and old (22 ‐ 24 months) males F344 rats were isolated and mounted on a pressure myograph system and changes in outer diameter in response to alteration of intravascular pressure (0‐100 mmHg) with a 20 mmHg increments under no flow condition were recorded. Myogenic tone calculated as a percentage of the difference between Active (in the presence of calcium) and Passive (in the absence of calcium) diameter over passive diameter for each pressure point. Vessel sizes determined as active outer diameter at 80 mmHg. Values calculated using Student t‐test and expressed as (mean ± s.e.m.). Myogneic tone calculated from arteries of young rats 115 µm ± 9 n= 12 (diameter range; 75 ‐ 167 µm) was significantly higher compared to old one 125 µm ± 9 n = 14 (diameter range; 72 ‐ 186 µm) (p蠄0.001). However, when subdividing vessel sizes into small arteries for young (88 µm ± 4) n= 6 and old (92 µm ± 5) n = 6; and large arteries for young (142 µm ± 6) n= 6 and old (149 µm ± 8) n = 8, the myogenic tone was highly significant when comparing small arteries from young and old (p<0.001) and there was no significant differences in myogenic tone between large arteries. Study findings stating that; Both myogenic tone and response decreased dramatically with aging in small resistance arteries, while increasing in young one. There is a very narrow range of myogenic tone drop, so care must be taken when selecting vessels sizes range for aging studies.

Grant Funding Source: Supported by Sultan Qaboos University, Oman
Endothelial cell‐targeted over‐expressing ET‐1 leads to increased blood pressure and decreased heart rate in aged mice (LB655)
Authors: Luo Xin , Xia Zhengyuan , Chung Sookja , Cheung Chi Wai
Published: June 2021

Endothelin‐1 (ET‐1) is a 21 amino acid vasoconstrictor and implicated in the regulation of hypertention. Recently, our colleagues found that transgenic mice (TET‐1) with endothelial ET‐1 over‐expression developed hypertention in young mice by changing vascular properties. However, the effects of over‐expressing endothelial ET‐1 on cardiovascular system in aged mice are still unclear. Therefore, this study investigated the blood pressure and heart rate of TET‐1 aged mice and non‐transgenic (NTg) aged mice (1.5‐2 years old). A non‐invasive tail cuff measurement showed that a significant elevation of both systolic (115.1 ± 2.935 vs 103.5 ± 1.846 mmHg, p<0.01) and diastolic (85.81 ± 2.303 vs 74.56 ± 2.029 mmHg, p<0.05) in conscious TET‐1 aged mice comparing to NTg counterpart, indicating over‐expressed endothelial ET‐1 cause both systolic and diastolic hypertension in aged mice. And, aged TET‐1 mice showed a significantly decreased heart rate to aged NTg mice (616.7 ± 20.14 vs 670.9 ± 11.57 beats per minute (bpm), p<0.05). Moreover, Rate‐pressure product is a sensitive index of myocardial oxygen consumption, and it was not altered in TET‐1 mice (71085 ± 3258 vs 69337 ± 1233 bpm*mmHg, p>0.05), implicating over‐expressed endothelial ET‐1 would change cardiovascular properties by mechanisms of mutual compensation between blood pressure and heart rate. These findings provide evidence for roles of endothelial ET‐1 in altering cardiovascular system in aged mice. This study also implicates clinical importance of endothelial ET‐1 as a therapeutic target for hypertension in the elderly.

Grant Funding Source: Supported by the departmental fund, Department of Anaesthesiology, The University of Hong Kong
Increased β‐adrenergic inotropy in ventricular myocardium from trpm4‐/‐ mice. (LB666)
Authors: Mathar Ilka , Kecskes Miklos , Van der Mieren Gerry , Jacobs Griet , Camacho Londoño Juan E. , Uhl Sebastian , Flockerzi Veit , Voets Thomas , Freichel Marc , Nilius Bernd , Herijgers Paul , Vennekens Rudi
Published: June 2021

RATIONALE: The Trpm4 gene has recently been associated with several disorders, including cardiac conduction diseases and Brugada syndrome. Transient receptor potential member 4 (TRPM4) proteins constitute Ca(2+)‐activated, but Ca(2+)‐impermeable, nonselective cation channels and are expressed both in atrial and in ventricular cardiomyocytes. The physiological function of TRPM4 in the heart remains, however, incompletely understood.

OBJECTIVE: To establish the role of TRPM4 in cardiac muscle function.

METHODS AND RESULTS: We used TRPM4 knockout mice and performed patch‐clamp experiments, membrane potential measurements, microfluorometry, contractility measurements, and in vivo pressure‐volume loop analysis. We demonstrate that TRPM4 proteins are functionally present in mouse ventricular myocytes and are activated on Ca(2+)‐induced Ca(2+) release. In Trpm4(‐/‐) mice, cardiac muscle displays an increased β‐adrenergic inotropic response both in vitro and in vivo. Measurements of action potential duration show a significantly decreased time for 50% and 90% repolarization in Trpm4(‐/‐) ventricular myocytes. We provide evidence that this change in action potential shape leads to an increased driving force for the L‐type Ca(2+) current during the action potential, which explains the altered contractility of the heart muscle.

CONCLUSIONS: Our results show that functional TRPM4 proteins are novel determinants of the inotropic effect of β‐adrenergic stimulation on the ventricular heart muscle.
Rats with hypertension, hemolysis, elevated liver enzymes and low platelets (HELLP) have increased placental and circulating FASL (LB713)
Authors: Wallace Kedra , Mathis Keisa , Bowles Teylor , Spencer Shauna‐Kay , Morris Rachael
Published: June 2021

The CD95L or FASL system of apoptosis is part of the tumor necrosis factor family that contributes to cell death. Placental CD95L is up‐regulated in women with HELLP syndrome and has been proposed to contribute to the liver damage seen in these patients. The objective of the current study was to determine if animals with HELLP have increased CD95L similar to women with HELLP. Additionally, as the anti‐inflammatory drug dexamethasone has been found to decrease inflammation in women with HELLP we also examined its’ effects on circulating CD95L in women with HELLP. Whole blood was collected from women with HELLP before and 24hrs after dexamethasone administration (10mg per 12hr intravenously). On gestational day (GD) 12, sEndoglin (7ug/kg/day) and sFlt‐1 (4.7ug/kg/day) infusion began via mini‐osmotic pump into normal pregnant (NP) rats. On GD18 carotid catheters were inserted, GD19 plasma and tissue were collected and CD95L was measured via western blot or enzyme linked immunosorbent assay. Women with HELLP had significantly higher levels of circulating CD95L compared to NP women (132.7+35 vs. 29.11+16; n=6;P=0.02). HELLP rats had significantly more circulating CD95L compared to NP rats (179.6+24 vs. 117.1+13pg; n=5; P=0.05). HELLP rats also had increased production of placental CD95L as measured via western blot (17.97+3 vs. 9.45+2.7 O.D.). These data suggest that similar to women with HELLP syndrome, rats with HELLP syndrome have increased circulating CD95L. Future studies will examine the effects of dexamethasone on decreasing CD95L production.