bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic hemoglobin disorders and anemia in Cambodian women of reproductive age (389.7)
Authors: Karakochuk Crystal , Whitfield Kyly , Talukder Aminuzzaman , McLean Judy , Green Timothy
Published: June 2021

Anemia is common in Cambodian women of reproductive age. Anemia can lead to adverse pregnancy outcomes and other serious health consequences. Potential causes of anemia include micronutrient deficiencies, infection and disease. In Cambodia, genetic hemoglobin (Hb) disorders are common, leading to anemia and other complications.

Objective: To examine associations between anemia (Hb <120g/L) and genetic Hb disorders in Cambodian women (18‐45 y).

Methods: 450 women from Prey Veng province provided blood. A complete blood count was performed and the presence and typing of hemoglobin disorders was carried out by PCR and electrophoresis.

Results: Overall, the prevalence of anemia in women was 33.1%, of which 61% was microcytic (Hb <120g/L and MCV <80fl). The prevalence of genetic hemoglobin disorders was over 55% (most commonly alpha‐thalassemia, Hb E and Hb CS) and were significantly correlated with anemia. Of women with anemia, over two‐thirds had an abnormal genetic hemoglobin type. Less than 5% of women had a low serum ferritin (< 15 µg/L).

Conclusion: The majority of anemia in Cambodian women is microcytic anemia. A major predictor of anemia was genetic Hb disorders. Low serum ferritin was uncommon suggesting that iron deficiency is not a major problem. However, genetic Hb disorders may be confounding the interpretation of ferritin and leading to an underestimation of iron‐deficiency anemia. Other indicators of iron status and other micronutrients involved in anemia are being examined.

Grant Funding Source: Supported by The International Development Research Centre and The Department of Foreign Affairs, Tr
Simplified approaches for estimating vitamin A stores and β‐carotene bioconversion in humans (39.6)
Authors: Park Hyunjin , Lietz Georg , Oxley Anthony , Boddy Alan , Berry Phil , Wyss Adrian , Schalch Wolfgang , Green Michael
Published: June 2021

Better methods are needed to assess vitamin A (VA) status and the efficiency of bioconversion of β‐carotene (BC) to retinol (ROH). Data on plasma ROH kinetics from 2 h to 14 d after an oral tracer dose of [13C10]BC and [13C10]retinyl acetate (RAc) to 33 healthy young adults were analyzed using model‐based compartmental analysis (WinSAAM, the Windows version of the Simulation, Analysis and Modeling software). The 6‐compartment model that fit data for all subjects predicted 5.3 pools of plasma ROH were transferred into extravascular stores each day, with ~17%/d recycling back to plasma; 6.5%/day was irreversibly lost and total body VA stores (TBS) were 146 ± 89 μmol (mean ± SD). We derived a simplified isotope dilution (“Olson”) equation for TBS, eliminating several factors and assumptions: TBS = F * (1 / SA), where F (fraction of dose [FD] absorbed and retained) was estimated as 0.61 from the kinetic data and SA (specific activity) is FD 13C10[ROH] in plasma 3 d after dosing / plasma ROH pool (μmol). TBS calculated using the equation was 149 ± 85 μmol, essentially the same as the value predicted by the model. BC bioconversion, calculated as FD 13C5[ROH] (derived from BC) / 13C10[ROH] (derived from RAc) at 2 d, averaged 23 ± 11% and was significantly correlated (R=0.942) with WinSAAM’s estimate based on areas under the curves (26 ± 12%). Our results indicate that both TBS and BC conversion to ROH can be estimated based on blood samples taken 3 and 2 d, respectively, after dosing. With further refinement, one sample at 3 d may suffice.

Grant Funding Source: BBSRC, U.K. and DSM, Basel, Switzerland
Child‐directed marketing within and around fast‐food restaurants (390.1)
Authors: Ohri‐Vachaspati Punam , Zeynep Isgor , Rimkus Leah , Powell Lisa
Published: June 2021

Fast food is the second‐largest source of calories among youth. Children who eat fast food consume more calories and have poor diet quality. The fast‐food industry spends over $700 million to market products to youth each year. Using validated instruments, data were collected from 6710 fast‐food restaurants located in a nationally representative sample of public middle‐ and high‐school enrollment areas between 2010 and 2012. Child‐directed marketing was defined as the presence of one or more of eight components inside or on the exterior of the restaurant. Over a fifth of all types of fast‐food restaurants and a third of chain fast‐food restaurants used child‐directed marketing within and/or around their buildings. Fast‐food restaurants that offered kids’ meals were significantly more likely to use child‐directed marketing than those that didn’t offer kids’ meals. Indoor display of kids’ meal toys was the most often used strategy and prevalence was highest among restaurants that participate in the Children’s Food and Beverage Advertising Initiative (CFBAI [84.6% in 2012]). Results from multivariate analysis indicate that all fast‐food restaurants in mid‐low income neighborhoods, and CFBAI member fast‐food restaurants in black neighborhoods, had significantly higher odds of using child directed marketing compared to those located in highest income and white neighborhoods, respectively. Child‐directed marketing was more prevalent in communities that are disproportionately affected by obesity. Popularity of kids’ meals toy displays to advertise to children meals that have been shown to be of poor nutritional quality is of concern. It is important to limit children’s exposure to unhealthy food advertising.

Grant Funding Source: Robert Wood Johnson Foundation
A comparison of the nutritional quality of kids’ versus adult meals from chain sit‐down restaurants (390.2)
Authors: Scourboutakos Mary , L'Abbe Mary
Published: June 2021

The objectives were to compare the nutritional quality of restaurant meals from regular (adult) menus in contrast to meals from kids’ menus, and to compare the nutritional quality of meals marketed as being “healthy kids’ meals” in contrast to “healthy adult meals”. Nutrition information was collected from Canadian chain restaurant websites in 2010. Nutrient levels (including calories, sodium, sugar, saturated fat, fiber, protein and trans fat) and Ofcom nutrient profile scores in 1630 adult meals and 248 kids’ meals from seven restaurant chains were analyzed. In addition, 46 “healthy kids’ meals” and 20 “healthy adult meals” were compared. On average, kids’ meals had a significantly lower nutritional quality in comparison to adult meals (p<0.01). However, the difference varied depending on the restaurant. “Healthy kids’ meals” had significantly higher amounts of calories, sugar, trans and saturated fat (both per serving and per 100g, p<0.05) when compared to “healthy adult meals”. Additionally, “healthy kids’ meals” had two‐times more saturated fat, almost three‐times more sugar and a significantly higher percentage of total energy coming from fat (33% [healthy kids’ meals] vs 25% [healthy adult meals]), saturated fat (11% vs 7%) and sugar (23% vs 11%). In conclusion, kids’ meals are often nutritionally worse than adult meals, and there is a gap between what is being marketed as a “healthy meal” for kids versus adults.

Grant Funding Source: Supported By: Vanier Canada Graduate Scholarship, CIHR/HSF PICDP Fellowship, OGS, McHenry Grant
Front‐of‐pack symbols are not a reliable indicator of products with healthier nutrient profiles (390.7)
Authors: Emrich Teri , Qi Ying , Cohen Joanna , Lou Wendy , L'Abbe Mary
Published: June 2021

Front‐of‐pack (FOP) nutrition rating symbols are used on food labels worldwide. Without standardized criteria for their use, it is unclear if FOP symbols are being used to promote products that are more nutritious than products without symbols. Objective: To compare the amount of calories, saturated fat, sodium, and sugar in products with FOP symbols to products without symbols. Design: A 2010‐2011 database of 10,487 Canadian packaged food labels was used. Nutrient content differences were compared using Wilcoxon rank‐sum test; differences greater than 25% were deemed nutritionally relevant. Results: Products with FOP symbols were not uniformly lower in calories, saturated fat, sodium, and sugar per reference amount than products without these symbols in any of the 10 food categories or in 59/60 subcategories. None of the different FOP system types (nutrient‐specific, summary indicatory, or food group information) examined were used to market products with overall better nutritional profiles (i.e. lower in calories, saturated fat, sodium, and sugar) than products without this type of marketing. Conclusion: FOP symbols are being used to market foods that are no more nutritious than foods without this type of marketing. As FOP symbols may influence consumer product perceptions and purchases, it may be a useful public health strategy to set minimum nutritional standards for products using FOP symbols.

Grant Funding Source: Canadian Institutes of Health Research
Contribution of food groups to the total dietary energy intake of Mexican children, adolescents and adults (393.3)
Authors: Pedraza Lilia , Aburto Tania , Sánchez Tania , Rivera Juan
Published: June 2021

OBJECTIVE. To study the contribution of food groups to the total dietary energy intake of a representative sample of Mexican children, adolescents and adults. METHODS. Dietary intake data from a random sample of children, adolescents and adults of the National Health and Nutrition Survey 2012 (ENSANUT 2012) was obtained using a single 24h recall. Seven food groups (FG) were considered: 1) cereals, 2) legumes and nuts, 3) milk and dairy, 4) meat, poultry, fish, eggs and processed meats, 5) fruit and vegetables, 6) sugar sweetened beverages (SSB) and 7) high energy density foods, high in sugar or fat (HEDF) such as candies, chocolates, chips, deserts, cakes and fast food. FG 1‐5 were considered indispensable foods while FG 6‐7 dispensable. All analyses were adjusted for the survey design. RESULTS. Percent contributions for children < 5y, 5‐11y, adolescents and adults, respectively, were: 22, 30, 32 and 35 for FG1; 3, 4, 3 and 4 for FG2; 24, 14, 11 and 9 for FG3; 11, 13, 15, 14 for FG4; 7, 5, 4, and 5 for FG5 (about 240 g); 6, 7, 9 and 12 for FG6; and 18, 18, 16 and 11 for FG7 (males). Results for females were similar in general. The dispensable FGs provided around 25% of the total energy intake (9‐12% from SSB and 11‐18% from HEDF) and the intake of fruits and vegetables was close to 60% of recommendations. CONCLUSIONS. Given that about 70% of adults and 33% of children are overweight or obese in Mexico, it is recommended that the intake of dispensable food groups should be reduced.
The landscape of viral expression and host gene fusion and adaptation in human cancer (397.5)
Authors: Tang Ka‐Wei
Published: June 2021

Viruses cause 10‐15% of all human cancers. Massively parallel sequencing has recently proved effective for uncovering novel viruses and virus‐tumour associations, but this approach has not yet been applied to comprehensive patient cohorts. Here we screen a diverse landscape of human cancer, encompassing 4,433 tumours and 19 cancer types, for known and novel expressed viruses based on >700 billion transcriptome sequencing reads from The Cancer Genome Atlas Research Network. The resulting map confirms and extends current knowledge. We observe recurrent fusion events, including human papillomavirus insertions in RAD51B and ERBB2. Patterns of coadaptation between host and viral gene expression give clues to papillomavirus oncogene function. Importantly, our analysis argues strongly against viral aetiology in several cancers where this has frequently been proposed. We provide a virus‐tumour map of unprecedented scale that constitutes a reference for future studies of tumour‐associated viruses using transcriptome sequencing data.
Mathematical modeling and correlation of histopathology with perfusion‐contrast CT for prediction of vascular permeability and chemotherapeutic diffusion parameters (397.7)
Authors: Bearer Elaine , Wang Zhihui , Cristini Vittorio
Published: June 2021

Tumor perfusion influences the ability of chemotherapeutic drugs to penetrate and kill cancers. Further, the physical properties of the microenvironment moderates the penetration of drugs from the vasculature into tumors. Here, we develop a mathematical model based on histopathology to predict the outcome of chemotherapy based on the physical laws of diffusion. The most important parameters in the model are the volume fraction occupied by tumor blood vessels (BVF) and their average diameter. Drug delivery to cells, and kill thereof, are mediated by these micro‐environmental properties and affected by the diffusion penetration distance after extravasation. To calculate parameter values we fit the model to histopathology measurements of the fraction of tumor killed after chemotherapy in human patients with colorectal cancer (CRC) metastatic to liver (coefficient of determination R2 = 0.94). To validate the model in a different tumor type, we input patient‐specific model parameter values from glioblastoma (GBM); the model successfully predicts extent of tumor kill after chemotherapy (R2 = 0.7‐0.91). Towards prospective clinical translation, we calculate BVF parameter values from in vivo contrast‐CT perfusion imaging from a separate cohort of patients with CRC metastatic to liver, and demonstrate accurate model predictions of individual patient responses (average relative error = 15%). Here, patient‐specific data from either in‐vivo imaging or histopathology drives output of the model’s formulas. Values obtained from histopathologic analysis for each individual are entered into the model dmonstrating accurate prediction of tumor kill after chemotherapy in a retrospective study of 24 subjects. Application of this model will enable rational design of individualized treatment strategies such as amount, frequency and delivery platform of drug and the need for ancillary non‐drug‐based treatment.

Grant Funding Source: NCI CA153825, NIGMS P50GM085273, NINDS NS062184
COX‐2 is induced by the COX‐2 selective inhibitors celecoxib and etodolac and the non‐selective inhibitor ibuprofen in several human tumor cell lines (397.8)
Authors: Schneider Ryan , Miller Ian , Renz Mackenzie , Whited Tawna , Kim Lindsay , Adams Bryce , Dudley Richard , Kinder David
Published: June 2021

The COX‐2 selective inhibitor, celecoxib, causes cancer cell death in vitro. However, the mechanism(s) behind the observed cytotoxic effects are still being debated. We have demonstrated that celecoxib treatment induces caspase‐dependent apoptosis in several human tumor cell lines (A375 melanoma, HT‐29 colon carcinoma and CRL‐1620 glioblastoma). Given protective effects of certain prostaglandins, we hypothesized that the apoptotic effects of celecoxib may be in part due to COX‐2 inhibition. However, western blotting for COX‐2 in A375, HT‐29 and CRL‐1620 cells treated with celecoxib for 24 hours revealed a surprising result. Celecoxib induced COX‐2 levels in each of these lines in a dose‐dependent manner. To determine whether COX‐2 induction by celecoxib is a class effect, we included the COX‐2 selective inhibitor etodolac and the non‐selective COX inhibitor ibuprofen. Etodolac treatment for 24 hours induced COX‐2 in a dose‐dependent manner in A375 and CRL‐1620 cells and at the highest dose in HT‐29 cells. Treatment with ibuprofen for 24 hours induced COX‐2 in a dose‐dependent manner in only A375 melanoma cells. These results demonstrate that the COX‐2 inhibitors celecoxib and etodolac induce protein expression of COX‐2 in several human tumor cell lines, whereas induction by ibuprofen was only seen in A375 tumor cells. These data demonstrate that specific and non‐COX specific NSAIDs induce COX‐2 expression.
Blocking Wnt secretion from hepatocyte impairs repairs following DDC injury in mice (398.2)
Authors: Okabe Hirohisa , Yang Jing , Singh Sucha , Nejak‐Bowen Kari , Monga Satdarshan
Published: June 2021

Diethoxycarbonyl dihydrocollidine (DDC) diet induces significant bile duct and hepatocyte injury mimicking human cholestatic liver disease. Following DDC injury, repair is often observed via ductular proliferation of expansion of biliary markers in the hepatocytes. However, the role of Wnt secretion by hepatocytes during DDC injury and repair remains elusive. Alb‐Cre+/‐;Wlsflox/flox mice, which lack the ability to secrete Wnt from hepatocytes, were exposed to DDC diet, and pathological characteristics were compared between wild type (WT) and hepatocyte‐specific Wls knock out (KO) mice. Expression of Wnt7A, Wnt7B and Wnt10B were induced by DDC treatment in WT mice. WT had more abundant A6 positive ductular cells indicating higher ductular proliferation as compared to KO. WT showed more CD45+ cells and collagen deposits than KO. KO showed higher serum total bilirubin level, and poor survival, resulting in worse prognosis than WT. Wnt secreted by hepatocytes might be crucial factor to induce ductular proliferation. This response is important repair mechanism after DDC injury.
Hepatocyte‐derived exosomes regulate liver recovery and regeneration after ischemia/reperfusion in mice (398.3)
Authors: Nojima Hiroyuki , Wilson Gregory , Quillin Ralph , Schuster Rebecca , Blanchard John , Edwards Michael , Gulbins Erich , Lentsch Alex
Published: June 2021

Exosomes are small extracellular membrane vesicles that originated from a variety of different cell types, including hepatocytes. Exosomes may play important roles in intercellular communication. Our previous work has demonstrated that the chemokine receptors, CXCR1 and CXCR2, regulate liver recovery and regeneration after ischemia/reperfusion (I/R) injury. In the present study, we sought to determine if hepatocyte‐derived exosomes mediate the CXCR1 and/or CXCR2‐mediated regulation of liver recovery and regeneration after I/R injury. Exosome release from hepatocytes was found to be dependent upon CXCR1 and CXCR2. CXCR1‐/‐ hepatocytes produced fewer exosomes, whereas CXCR2‐/‐ hepatocytes produced more exosomes compared to their wild‐type controls. Exosomes from wild‐type hepatocytes induced dose‐dependent increases in hepatocyte proliferation in vitro. Interestingly, exosomes from CXCR1‐/‐ hepatocytes had no effect on hepatocyte proliferation, whereas exosomes from CXCR2‐/‐ hepatocytes had similar effects as those from wild‐type cells. Mice treated with hepatocyte‐derived exosomes showed a dose‐dependent increase in hepatocyte proliferation after I/R. The data suggest that exosome release by hepatocytes is controlled by CXCR1 and CXCR2 and that hepatocyte‐derived exosomes promote hepatocyte proliferation and liver regeneration after I/R injury.
Low‐fat dairy reduces inflammatory genes and liver enzymes in adults with metabolic syndrome (40.5)
Authors: AGuilar David , Dugan Christine , Park Young‐Ki , Lee Ji‐Young , Fernandez Maria Luz
Published: June 2021

We evaluated the effect of low‐fat dairy (LFD) intake on plasma inflammatory markers, gene expression of these markers in peripheral blood mononuclear cells (PBMC) and liver enzymes in individuals with metabolic syndrome (MetS). Subjects (13 male, 22 female) with low dairy intake (<1.5 ser/dairy/day) were randomly assigned to receive LFD (296 mL low‐fat milk, 170 g yogurt, 510 g cheese stick, 3 servings of dairy/day) or isocaloric control foods (CON) (42 g granola bar, 177 mL juice) daily for 6‐wk. After a 4‐wk wash‐out, subjects were allocated to the alternate treatment. Participants had lower concentrations of both hepatic alanine aminotransferase (ALT) (p < 0.05) and aspartate aminotransferase (AST) (p< 0.005) as well as lower concentrations of plasma monocyte chemoattractant protein (MPC‐1) (p<0.05) after the LFD period. Gene expression was evaluated in a subset of subjects (n = 17). IL‐1b and IL‐6 were reduced by 46 and 63%, respectively compared to the control period. Protein and gene expression of tumor necrosis factor (TNF)‐alpha was not altered by diet. These results indicate that consumption of three dairy servings per day improve both liver inflammatory markers and systemic inflammation in subjects with MetS.

Grant Funding Source: Dairy Council
lmpact of various dietary oils on expression levels of inflammatory genes: a randomized crossover controlled nutritional intervention (40.6)
Authors: Labonté Marie‐Eve , Couture Patrick , Vohl Marie‐Claude , Jenkins David , Connelly Philip , West Sheila , Kris‐Etherton Penny , Jones Peter , Lamarche Benoît
Published: June 2021

Consumption of a docosahexaenoic acid (DHA)‐enriched canola oil was shown to have anti‐inflammatory effects compared with a control diet rich in linoleic acid (LA) and a diet rich in flax oil and alpha‐linolenic acid (ALA). We further investigated the impact of these oils on the expression of key inflammatory genes in whole blood cells as part of the Canola Oil Multicenter Intervention Trial (COMIT). A total of 118 men and women with abdominal obesity and at least one other criterion for metabolic syndrome consumed 5 isoenergetic diets for 4 weeks each according to a randomized, crossover design. Each diet provided 60 g/3000 kcal of various experimental oils. Here we report the impact of 3 of these 5 oils: 1‐ CONTROL high LA corn/safflower oil (LA 41.6 g; ALA 0.2 g; n6/n3 ratio 208:1), 2‐ FLAX oil (LA 22.5 g; ALA 19.2 g; n6/n3 ratio 1.2:1), 3‐ High DHA canola (DHA‐CAN; LA 7.6 g; ALA 1.2 g; DHA 3.5 g; n6/n3 ratio 1.2:1). Inflammatory gene expression in whole blood cells at the end of each diet was assessed by real‐time PCR in a subset of 66 individuals. DHA‐CAN lowered mRNA expression of interleukin(IL)‐1β compared with CONTROL (fold change 0.90, P=0.03) and FLAX (fold change 0.89, P=0.03). There was no between‐diet difference in the expression of other inflammatory genes and transcription factors. These data suggest that anti‐inflammatory effects of DHA compared with n3 and n6 polyunsaturated fatty acid rich oils from plant sources occur at least in part through a reduction in the expression of IL‐1β.

Grant Funding Source: Supported by the Canola Council of Canada, Flax 2015 and Agriculture and Agri‐Food Canada
Vitamin D status positively associates with interleukin‐6 and tumour necrosis factor alpha in healthy young children (40.8)
Authors: Brett Neil , Agellon Sherry , Vanstone Catherine , Weiler Hope
Published: June 2021

Vitamin D status associates with immune function and duration of illness. The objective was to examine interleukin‐6 (IL‐6) and tumor necrosis factor alpha (TNF) in relation to vitamin D status in healthy children. Children (n=457; 1.8‐5.8 y) were recruited from randomly selected daycares in Montreal. Body mass index Z‐scores for age (BAZ) were calculated using the WHO growth standard. Capillary plasma samples were measured for IL‐6, TNF and C‐related peptide (CRP) (Multiplex assays; Millipore Corp. USA); CRP values > 10 mg/L were excluded. Vitamin D status (chemiluminescence; Liaison, Diasorin Inc.) was categorized using plasma 25‐hydroxyvitamin D (25(OH)D) values <50.0 nmol/L (low; n=52), 50.0‐74.9 nmol/L (adequate; n=189) and 蠅75.0 nmol/L (robust; n=216). Differences among vitamin D status categories were tested using mixed model ANOVA (random effects: age, sex, BMI Z‐score and season) and Tukey’s post hoc test; data are mean ± SD. Average age was 3.7 ± 1.0 y, BAZ 0.5 ± 1.0 kg/m2 and CRP 0.9 ± 1.6 mg/L. By design all pair‐wise differences in 25(OH)D (nmol/L) were observed (low: 40.8 ± 6.7; adequate: 63.4 ± 7.6; robust 100.3 ± 22.4, P<0.001). Highest IL‐6 (low: 9.9 ± 13.0; adequate 9.9 ± 12.9; robust 14.0 ± 14.9 ng/L, P<0.02) and TNF (low: 6.8 ± 3.5; adequate 7.1 ± 3.1; robust 7.8 ± 3.4 ng/L, P<0.03) were observed in the robust group. The data suggests that robust vitamin D status supports innate immune function.

Grant Funding Source: Supported by Dairy Farmers of Canada
CFC1 gene mutation in tetralogy of Fallot and dextro‐transposition of great arteries in Pakistani population. (404.2)
Authors: Arif Afsheen , Zehra Sitwat , Qamarunissa Syeda , Patel Najma , Azhar Abid
Published: June 2021

Background: Congenital heart diseases (CHDs) are the most common of all birth defects and one of the leading causes of mortality in the first year of life. CFC1 gene is a cell signaling protein is a co receptor in nodal signaling pathway and involve in right and left axis determination during gastrulation.

Materials and Methods: This is a case‐control study, recruited 225 non syndromic patients and 140 controls, healthy unrelated individuals. The study, after formal approval includes patients from various pediatric cardiology centers in three years. A detailed family history was taken to elucidate the genetic and environmental factors. Pediatric cardiologist confirmed the diagnosis on the basis of all standard testing like chest X‐ray, CBC, ECG, ECHO, cardiac catherization reports etc. DNA extraction and sequencing was done and data was interpreted by multiple sequence alignment software. Statistical data was done by SPSS 17.0.

Results: The mean age for controls was 3.14±1.82 years, for TOF; 2.97±1.21, and for DTGA patients 1.84±2.26 years. TOF and DTGA were frequent in males. The study demonstrates frequency of this disease; with its variation in Pakistani population. Consanguinity affects the rate of CHDs, as it is 62% in patients and 25% in controls. Two novel mutations were found in CFC 1 gene.First in exon 5 at c. 396 C>A and second in exon 6 at c. 548 C>T.

Conclusions: The study demonstrates frequency and prevalence for TOF, and DTGA, there variation and association with other cardiac defects. Environmental factors are taken into consideration and consanguinity proves to be an element for the increasing number of disease. CFC1 plays a key role in cardiac malformation and its mutations.
SQS promotes metastasis through altered cholesterol contents and recruiting TNFR1 to lipid rafts for NFκB activation in lung cancer (405.1)
Authors: Yang Yi‐Fang , Jan Yi‐Hua , Yang Chih‐Jen , Liu Yupeng , Su Chia‐Yi , Tsai Hong‐Yuan , Lai Tsung‐Ching , Shen Chia‐Ning , Shew Jin‐Yuh , Lu Pei‐Jung , Huang Ming‐Shyan , Hsiao Michael
Published: June 2021

Identifying biomarkers that predict patient outcome might provide opportunities for developing better therapeutic strategies to prolong metastatic lung cancer patient survival. The first enzyme of the sterol branch of cholesterol synthesis, squalene synthase (SQS), is associated with lipid rafts and upregulated in several cancer types. Here, loss of SQS expression significantly suppressed the invasion potential of lung cancer cells, whereas SQS overexpression promoted invasion and metastasis in vitro and in vivo. Similar effects occurred after treatment with the SQS chemical inhibitor zaragozic acid A in vitro and in vivo. SQS assists in the recruitment of tumour necrosis factor‐α receptor 1 (TNFR1) to lipid rafts, which activates NF‐κB. Further analysis revealed that matrix metalloproteinase 1 (MMP1) is an important mediator of SQS‐induced invasion. Overexpression of SQS resulted in enhanced tumour invasion/metastasis via lipid rafts‐mediated signalling and subsequent MMP1 upregulation. Targeting SQS may have great potential as a novel therapeutic strategy to treat metastatic lung cancer.
Caveolin‐1 deficiency enhances cancer cell transendothelial migration and promotes lung metastasis through upregulation of VCAM‐1 in lung endothelial cells (405.2)
Authors: Fu Wei , Wang Chen , Liu Ziyang , Wang Enhua , Liu Jun
Published: June 2021

Caveolin‐1 (Cav1) has been shown to play an important role in oncogene‐induced cell transformation and tumorigenesis. It has been reported that the level of Cav1 expression in tumors affected tumor cell proliferation, invasion and metastasis. In this study, we employed Cav1 knockout mice and tested whether genetic depletion of Cav1 in endothelial cells would affect cancer metastasis. Melanoma (B16F10) and Lewis lung cancer (LLC1) cells were inoculated in both wild‐type and Cav1 knockout mice. Tumor growth and metastasis in the lungs and livers were monitored for a period of four weeks. Although there was no significant difference of the onset and burden of the primary tumors between wild‐type and Cav1 knockout mice, Cav1 depletion significantly promoted tumor metastasis to the lungs and livers. Consequently, the survival rate was significantly lower in Cav1 knockout mice than the control group. Upon TNFα stimulation, Cav1 knockout endothelial cells showed more than 2‐fold increase in the expression of vascular cell adhesion molecule‐1 (VCAM‐1) over wild‐type cells. The increased VCAM‐1 expression was translated to a dramatic increase in the adhesion of B16F10 cells to Cav1 deficient endothelial cells by 5‐fold more than wild‐type cells. Furthermore, Cav1 deficiency resulted in an increase in the transendothelial migration of B16F10 cells. Our results suggest that the levels of Cav1 expression in endothelial cells plays important role in cancer metastasis by enhancing the interaction of cancer cells with endothelial cells and cancer cell extravasation.

Grant Funding Source: Supported by the American Cancer Society
Mice heterozygous for the F508del mutation in the cystic fibrosis transmembrane conductance regulator anion channel display attenuated cardiopulmonary dysfunction and lung injury after influenza H1N1 infection (406.4)
Authors: Aeffner Famke , Joseph Lisa , Amer Amal , Hickman‐Davis Judy , Davis Ian
Published: June 2021

Influenza A viruses cause a highly‐contagious respiratory disease associated with significant morbidity and mortality. The purpose of this study was to utilize F508del CFTR‐heterozygous mice to determine the contribution of CFTR‐mediated Cl‐ secretion to influenza pathogenesis and to evaluate CFTR as a therapeutic target.

F508del CFTR‐heterozygous progeny (HETs) and wild‐type (WT) littermate controls were infected intranasally with 50 μl virus diluent (mock) or with 10,000 FFU/mouse influenza A/WSN/33 (H1N1) for 1‐6 days. Body weight, lung mechanics, cardiopulmonary function and levels of pulmonary inflammatory mediators were determined over the time of disease.

Relative to WT littermate controls, infection‐induced hypoxemia and bradycardia were attenuated in HETs at 2‐6 days post‐infection (d.p.i.), although weight loss and viral replication did not differ. At 2 and 6 d.p.i., alterations in lung function in infected WT mice were absent in HETs at both 2 and 6 d.p.i. Bronchoalveolar lavage fluid from influenza‐infected HETs contained approximately 3‐fold greater numbers of alveolar macrophages and more than 10‐fold higher IL‐6 levels at 6 d.p.i.

Reduced CFTR expression resulting from F508del CFTR heterozygosity significantly alters influenza pathogenesis. CFTR‐mediated Cl‐ secretion is therefore a potential therapeutic target to decrease lung injury after influenza H1N1 infection.

Grant Funding Source: Supported by C. Glenn Barber Fund and NHLBI R01‐HL102469
SerpinA3N accelerates wound closure in a murine model of impaired wound healing (413.1)
Authors: Hsu Ivy , Zhao Hongyan , Bleackley Chris , Granville David
Published: June 2021

Granzyme B (GzmB) is a serine protease that accumulates in the extracellular milieu during chronic inflammation and cleaves many extracellular matrix (ECM) proteins that are essential for proper wound healing to ensue. We hypothesized that GzmB contributes to the pathogenesis of impaired wound healing observed in the diabetic population through excessive degradation of the ECM. On the dorsal back of genetically‐induced type II diabetic db/db mice, 10 mm excisional wounds were created and covered with semi‐transparent dressing. SerpinA3N, a GzmB inhibitor, was administered topically and/or subcutaneously on the wounds every 3 days. GzmB was found to co‐localize with mast cells in wound sites at the dermal‐epidermal junction. In addition, preliminary results suggest that administration of serpinA3N accelerates wound closure as by day 35, 100% of serpinA3N‐treated compared to 55% of vehicle‐treated mice have fully re‐reepithelialized. In serpinA3N‐treated animals, an increase in full‐length ECM proteins involved in collagen fibrillogenesis was observed along with fewer cleavage fragments by immunostaining. These findings suggest that GzmB contributes to the pathogenesis of diabetic wound healing through the proteolytic cleavage of ECM proteins that are essential for normal wound closure.

Grant Funding Source: Supported by Canadian Diabetes Association and Canadian Institutes of Health Research
Relevance of GPCR functional selectivity/biased signaling to drugs of abuse (468.1)
Authors: Caron Marc , Urs Nikhil , Peterson Sean , Daigle Tanya , Snyder Joshua
Published: June 2021

We now understand that GPCRs can signal not only through activation of G proteins but also through the ability of β‐arrestins to scaffold intracellular signaling molecules with distinct temporal and pharmacological properties. G protein‐ and β‐arrestin‐dependent signaling have been shown to mediate distinct cellular responses. We have previously demonstrated that both subfamilies of dopamine receptors, D1R and D2R, which are downstream targets of psychostimulant drugs of abuse, can signal through both G protein‐ and β‐arrestin‐mediated pathways. We have used genetic approaches to delete β‐arrestins globally or in selective neuronal populations to determine which signaling pathways mediate specific behavioral responses to various psychostimulants. Collectively, our data suggest that both G protein‐ and β‐arrestin‐dependent signaling play an important role in psychomotor responses whereas only G protein signaling downstream of dopamine receptors contributes predominantly to the reinforcing effects of psychostimulants. These findings suggest that developing functionally selective receptor‐based therapeutic approaches may represent a viable avenue to manage drugs of abuse.
Chemical genetics to map kinase signaling pathways (473.3)
Authors: Shokat Kevan
Published: June 2021

Kinases are highly regulated enzymes with diverse mechanisms controlling their catalytic output. Inhibitor discovery efforts for kinases have produced ATP‐competitive compounds, allosteric regulators, irreversible binders, and highly specific inhibitors. These distinct classes of small molecules have revealed many novel aspects about kinase‐mediated signaling and some have progressed from simple tool compounds into clinically validated therapeutics. My presentation will explore several small‐molecule inhibitors for kinases highlighting elaborate mechanisms by which kinase function is modulated. A complete surprise of targeted kinase drug discovery has been discovery of ATP‐competitive inhibitors that behave as agonists, rather than antagonists, of their direct kinase target. These studies hint at a connection between ATP binding site occupancy and networks of communication that are independent of kinase catalysis. Indeed, kinase inhibitors have been found that induce changes in protein localization, protein‐protein interactions, and even enhancement of catalytic activity of the target kinase. The relevance of these findings to the therapeutic efficacy of kinase inhibitors and to the future identification of new classes of drug targets will be discussed.
Regulation of neuron function and metabolism by acyl‐CoA hydrolysis (476.1)
Authors: Wolfgang Michael
Published: June 2021

Numerous neurological diseases are associated with dysregulated lipid metabolism; however, the regulation of fatty acid metabolism in neurons has been largely unexplored. The control of cellular fatty acid flux lies in the activation of fatty acids into acyl‐CoAs, and counterbalanced by the deactivation of acyl‐CoAs by acyl‐CoA thioesterases (ACOTs). ACOT7 is specifically and abundantly expressed in neurons, suggesting that it serves to antagonize fatty acid metabolism in neurons which have minimal capacity to oxidize or store lipids. To determine the role of ACOT7 in neural function and metabolism we have generated a nervous system specific ACOT7 knockout mouse, Acot7N‐/‐. Acot7N‐/‐ brains have increased fatty acid flux into complex lipid synthesis and increased phospholipid remodeling particularly under fasting conditions. The loss of neuronal ACOT7 causes physiological and behavioral abnormalities manifesting in a loss of loss of lean mass, hypermetabolism, hepatic steatosis, dyslipidemia, neurological dysfunction, neurodegeneration and behavioral hyperexcitability. The hyperexcitability of Acot7N‐/‐ mice is further exemplified by increased sensitivity to kainic acid induced seizures. These data show that ACOT7 mediated acyl‐CoA hydrolysis counter‐regulates fatty acid metabolism in neurons to protect against neurotoxicity and hyperexcitability.
Intestinal epithelial ecto‐5’‐nucleotidase CD73 regulates invasion by Salmonella typhimurium (488.4)
Authors: Kao Daniel , Saeedi Bejan , Vickery Thad , Wilson Kelly , Colgan Sean , Kominsky Douglas
Published: June 2021

Previous work has shown that adenosine is a key mediator of bacterial‐epithelial cell interactions. CD73 is an ecto‐5'‐nucleotidase expressed abundantly on the apical surface of intestinal epithelial cells (IEC) that catalyzes the conversion of 5’‐adenosine monophosphate (AMP) to adenosine (Ado). Here, we hypothesized that CD73 activity is important in invasion of IECs by the enteric pathogen Salmonella typhimurium. In this work, we demonstrate that pharmacologic inhibition of CD73 activity and lentiviral‐mediated knockdown of CD73 expression significantly attenuated Ado production by cultured T84 human colonic epithelial cells (approximately 7‐fold reduction in the ability to convert etheno‐AMP to etheno‐Ado). Furthermore, inhibition of CD73 activity by these two methods diminished invasion of T84 cells by Salmonella typhimurium. Based on these in vitro data, a transgenic mouse was generated with IEC‐specific knockout of CD73 (CD73f/f x Villin‐Cre). These CD73 tissue‐specific knockout mice were protected in an S. typhimurium model of infectious colitis compared to control animals. IEC‐specific knockout animals revealed attenuated weight loss and diminished histopathological severity of colitis at 24 and 48 hours after infection. Correspondingly, there was a significant decrease in disseminated bacteria isolated from mesenteric lymph nodes and spleen, demonstrating attenuated bacterial invasion in the absence of CD73. In conclusion, these results demonstrate that CD73 is a key regulator of infection by the enteric invasive pathogen S. typhimurium and implicates a prominent role for extracellular adenosine in Salmonella interactions with IEC.

Grant Funding Source: Supported by NIH and CCFA
Health Science & Technology Academy: an example of successful outreach and state infrastructure (531.23)
Authors: Klinkhachorn Penprapa , Radow Mark
Published: June 2021

HSTA is a WV program to increase health awareness, increase community engagement, and decrease health disparity. The goal of HSTA is to expose disadvantaged high school students to the STEM subjects to pursue a degree in health, science, and technology. The main programs offered are Fun with Science, Forensic Science, Biomedical Summer Institute, and Senior Preparation. Eligible students are African American (30%), low income (63%), females (67%), the first in their family to go to college (74%), and living in rural areas (83%). Anatomy is one of seven biomed groups offered, and the activities are fun, engaging, and interactive. The majority of the student survey responses were very positive. 94% of students learned a lot from their group inquiry and from visiting the gross lab. 75% agreed that HSTA was able to influence their decision to go to college and 84% to choose health careers. 82% are satisfied with the overall program. 97% of HSTA students go to college vs. 56% of all WV high school students. The gross lab tour was popular and the props used for heart and lungs helped to stimulate critical thinking skills. They learned the proper techniques to check vital signs and perform physical exams. Students had fun playing a Jeopardy game which was used as a tool to test what they learn that week. In conclusion, students participating in WVU HSTA Biomed Program learn a lot from hands‐on activities and competitive games in anatomy show and tell. This NIH‐funded infrastructure has been successful in providing opportunities for participating youths and in increasing the number of health science professionals in WV. Under supervision and guidance, these high school students can conduct activities to help increase awareness and better the overall health of their families and communities.

Grant Funding Source: Supported in part by NIH‐NCCR # 10009490.28.1003654R.
Using chick forebrain neurons to model neurodegeneration and protection in an undergraduate neurophysiology laboratory course (531.5)
Authors: Burdo Joseph
Published: June 2021

Since 2009 at Boston College, we have been offering a Research in Neuroscience course using cultured neurons in an in vitro model of stroke. The students work in groups to learn how to perform sterile animal cell culture and run several basic bioassays to assess cell viability. They are then tasked with analyzing the scientific literature in an attempt to identify and predict the intracellular pathways involved in neuronal death, and identify dietary antioxidant compounds that may provide protection based on their known effects in other cells. After each group constructs a hypothesis pertaining to the potential neuroprotection, we purchase one compound per group and the students test their hypotheses using a commonly performed viability assay. The groups generate quantitative data and perform basic statistics on that data to analyze it for statistical significance. Finally, the groups compile their data and other elements of their research experience into a poster for our departmental research celebration at the end of the spring semester.
An anatomical approach to teaching the clinical evaluation of musculoskeletal injuries to primary care providers (536.5)
Authors: Stamm Julie , Zumwalt Ann
Published: June 2021

A solid understanding of anatomy is essential for successful clinical evaluation of musculoskeletal injuries. However, reinforcement of basic anatomical knowledge is often not part of continuing medical education (CME) for providers in the primary care setting, a group that is often first to evaluate musculoskeletal injuries. To address this gap, two lectures, one on the knee and the other on the shoulder, were presented to primary care physicians, nurse practitioners, registered nurses, and medical assistants at a university student health center for CME credit. In pre‐lecture surveys the clinicians indicated that, on average, they were not confident to somewhat confident in their ability to evaluate knee and shoulder pain. A key feature of the lectures was their organization following the clinical examination process, with emphasis on palpation and testing of the underlying anatomy and differential diagnosis of common injuries. In post‐lecture surveys the clinicians reported increased knowledge of anatomy, palpation, testing procedures, and specific injuries as well as increased confidence in their evaluation of knee and shoulder injuries. This project demonstrates that basic science presentations for primary care clinicians given within the familiar context of the clinical examination can contribute to increasing their clinical knowledge and confidence with common musculoskeletal assessments.

Grant Funding Source: Supported by NIH NRSA Fellowship Award NINDS F31 NS081957
Ultrasound of femoral nerve: a gender comparison (536.9)
Authors: Benninger Brion , Abdulla Maha
Published: June 2021

INTRODUCTION: Femoral nerve blocks are common procedures in peri‐operative and acute pain settings. Limited research has been conducted on the factors which lead to incomplete nerve block or inadequate analgesic effect in some individuals. The objective of this pilot study was to investigate point of nerve arborization in males and females as a factor in this variability, and determine whether a trend exists. METHODS: Literature search was conducted on anatomy texts, atlases, and journals regarding femoral nerve morphology and gender variability. Femoral nerves of live subjects (20F‐40sides:13M‐26sides) were visualized with ultrasound and measured from the inguinal ligament to point of bifurcation. RESULTS: Literature search revealed anomalous branching patterns and mapping of the femoral nerve. No comparison of morphological female to male discrepancies was found. Ultrasound imaging indicated an average difference of approximately 1cm between genders. Arborization in males was more distal to the inguinal ligament than females. CONCLUSIONS: Optimal placement of femoral nerve blocks is imperative and may need to be modified based on patient gender to achieve maximum anesthesia. Adjusting injection sites for male and female patients could avoid the need for higher anesthetic volumes and may reduce occurrence of incomplete blocks. This study reveals a gender difference of the femoral nerve morphology, which suggests a further study to formally develop standardized protocols for each gender.
Wine and juice and oral cavity morphometric evaluation experimental study (54.1)
Authors: Costa Eduardo , Rodrigues Tiago , Cavalcanti Karen , Farinha Rodrigo , Cabrita Silvério
Published: June 2021

The foods used for feeding, as well as the way they’re cooked and mixed in the meals, are elements of great importance to the health of populations.舲 The aim of this study is to evaluate the existence of morphological changes in the periodontal ligament after administration of red wine and an extract of Genipa americana.

In this study were used 30 Wistar male rats with eight weeks at baseline, kept under physiological conditions of maintenance and handling, in accordance with the legislation in force. The Wistar male rats were distributed into three groups of 10. Group I was the control group continued for 8 weeks with no manipulation. The Group II was submitted to the administration of mature red wine, five times per week, for 8 weeks at a dose of 3.57 ml / kg each takes, and Group III was subjected to 8 weeks administration of an extract of G. americana. The extract was prepared by carrying out the juice of the fruit, the husk and seeds.

All animals were sacrificed and necropsy was made at the end of the administration.舲 In group I the ratio between the hyalinization areas and the total ligament area (R1) was 0.017 ± 0.015 and the ratio of the Tooth‐hard tissue area and periodontal ligament area (R2) was 0.896 ± 0.497. In group II the value of R1 was 0.03 ± 0.01 and the R2 of 0.87 ± 0.49. In group III the value of R1 was 0.03 ± 0.03 varying from 0 to 0,129 and R 2 0.83 ± 0.3, ranging from 0.39 and 1.63.

According to these results the administration of wine interferes with the tooth structure reflecting in the periodontal ligament and administration of genipapo doesn’t have a significant action on the dental structures studied.
Bacterial lipopolysaccharide inhibits VEGFR2 expression and cell migration in fetal mouse lung mesenchyme (540.1)
Authors: Greer Rachel , Prince Lawrence
Published: June 2021

Infection and inflammation inhibit normal lung development. We have previously measured multiple alterations in growth factor expression and signaling that may link inflammation with abnormal development. Here we report data investigating the effects of the bacterial endotoxin lipopolysaccharide (LPS) on developmental gene expression in fetal mouse lung mesenchyme. Three separate mesenchymal cell lines were established from the embryonal lungs of ImmortoMice at E11, E15, and E18. We used these lines to measure persistent changes in gene expression (using Affymetrix MoGene 1.0st arrays) following LPS treatment. While each line showed similar acute inflammatory responses to LPS, only a small subset of 285 genes was down regulated. Further analysis identified a smaller set of 9 genes that consistently decreased in all lines tested, including the VEGF receptor VEGFR2. While mesenchymal cells can transdifferentiate into endothelial cells, they also migrate along newly formed blood vessels to form vascular pericytes. Treating mesenchymal cells with LPS reduced VEGF‐induced cell migration, but did not decrease the response to PDGF. These data suggest that persistent inflammation may alter mesenchymal cell biology in the developing lung by altering normal VEGFR2 expression and disrupting the spatial and temporal relationships required for normal alveolar capillary formation.