bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro Analysis of Oligodendrocyte Differentiation Using Sox10‐Venus Mice (LB239)
Authors: Sekimoto Kaori , Suzuki Nobuharu , Mabuchi Yo , Akazawa Chihiro
Published: June 2021

In vitro Analysis of Oligodendrocyte Differentiation Using Sox10‐Venus Mice Kaori Sekimoto, Nobuharu Suzuki, Yo Mabuchi, Chihiro Akazawa: Depart. of Biochem. and Biophys., Grad. Sch. of Health Care Sci., TMDU, Tokyo, Japan Oligodendrocytes form myelin in the central nervous system (CNS) utilizing their highly branched cellular processes. However, detailed mechanisms of oligodendrocyte differentiation, including the process formation are poorly understood, particularly due to the difficulty of the purification of murine oligodendrocyte precursor cells (OPCs). We have recently established a transgenic mouse line that expresses a fluorescent protein Venus under the promoter of Sox10, whose expression is restricted to OPCs and oligodendrocytes. Here, we have characterized CNS Sox10‐Venus cells for analyzing oligodendrocyte differentiation. We first purified the Sox10‐Venus cells from the postnatal day 2 cerebral cortex by fluorescence activated cell sorting (FACS). Most of Sox10‐Venus‐positive cells expressed NG2, an OPC marker, although Sox10‐Venus‐negative cells did not. After induction of oligodendrocyte differentiation, an increased population of galactocerebroside (GalC)‐positive oligodendrocytes and decrease of OPCs were observed in the Sox10‐Venus‐positive culture, suggesting that the purification of OPCs, which are differentiable to oligodendrocytes, were succeeded using the Sox10‐Venus system. Furthermore, a time‐lapse analysis showed that Sox10‐Venus oligodendrocytes dynamically changed their morphology with highly branched cell processes during differentiation. In addition, we found that Sox10‐Venus OPCs were able to differentiate to type II astrocytes, which were previously found in normal OPC culture after an expose with a high serum concentration. Taken together, the Sox10‐Venus culture system is useful for analyzing the differentiation and cell process formation of oligodendrocytes and the multipotency of OPCs in mice.
Effect of cytochrome c modification with co‐polymer on its apoptotic activity for cancer treatment (LB248)
Authors: Figueroa Miranda Cindy , Morales‐Cruz Moraima , Suarez Bethzaida , Griebenow Kai
Published: June 2021

Treating cancer, while avoiding devastating side effects, has been a challenge throughout the years. Active targeted delivery has been extensively studied to overcome the inherent adverse effects of anticancer therapy. The apoptotic protein cytochrome c (cyt c) is highly effective at causing cancer cells cytotoxicity but as most proteins, its delivery is challenging. In this project, we synthesized a co‐polymer of hyaluronic acid (HA) and folic acid (FA) followed by cyt c binding. Both compounds of the co‐polymer were selected due to their receptors overexpression on cancer cells surface allowing endocytosis of the cyt c system. We used TNBSA assay to determine the amount of modified lysines in cyt c when reacted to HA‐FA: 2.1 ± 0.4. The residual activity of the protein was measured before and after its incorporation in HA‐FA using a caspase cascade assay which showed caspase activation by cyt c‐HA‐FA: 105 ± 1 % while HA‐FA did not activate caspase: 1.6 ± 0.8 %. CD spectroscopy showed minor changes in heme group and tertiary structure of cyt c following modification. HeLa cells were incubated with cyt c‐HA‐FA (0.1 mg/ml) to measure cell viability with MTS assay. Cell viability was measured after 2 and 6 hours of incubation: 108 ± 14 % and 83 ± 8 %, respectively. According to these results we successfully synthesized a promising drug delivery system for anticancer applications.

Grant Funding Source: Supported by NIH Research Initiative for Scientific Enhancement (RISE) Program with grant number: 2R25GM061151‐12.
Effects of 6‐hydroxynicotinate 3‐monooxygenase gene knockout (∆nicC) on nicotinate degradation and virulence modulation in Bordetella pertussis (LB266)
Authors: Chiaranunt Pailin , Sprano Jacob , Daniels Abigail , Snider Mark
Published: June 2021

Due to increased incidence of whooping cough, further investigation of the pathogenic mechanisms of its causative agent, Bordetella pertussis, is of medical importance. Previous in vitro assays indicate a correlation between elevated nicotinate concentrations and the down‐regulation of virulent toxins (e.g. pertussis toxin and adenylate cyclase) in B. pertussis. Furthermore, recent genomic analyses reveal a unique nicotinate degradation pathway in B. pertussis. This study analyzes the effects of disrupting the gene that codes for 6‐hydroxynicotinate 3‐monooxygenase (NicC), the enzyme predicted to catalyze the committed step in this pathway. A nicC knockout strain (ΔnicC) of B. pertussis Tohama I was created by insertion of a chloramphenicol‐resistance (CmR) gene via allelic exchange. Analyses by HPLC assays and RT‐PCR of nicotinate degradation and the resulting effects on virulence modulation, respectively, in ΔnicC and the wildtype Tohama I strains of B. pertussis are ongoing.
Analysis of the validity of a carbohydrate intolerance questionnaire II (LB304)
Authors: Goodenough C , Levers K , Galvan E , Coletta A , Dalton R , Jung Y , O'Connor A , Simbo S , Seesselberg C , Bonin B , Koozehchian M , 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, completed a CI, had body composition and health measures determined, and underwent a 75g, 2‐h OGTT. Having food cravings relieved with carbohydrate ingestion correlated with irritability (r=0.24), nervousness (r=0.32), forgetfulness (r=0.31), mental confusion (r=0.23), worrying (r=0.21), antisocial behavior (r=0.24), lack of sex drive (r=0.27), leg cramps/blurred vision (r=0.25), cravings for sweets (r=0.67), digestive disturbances (r=0.20), yawning (r=0.26), drowsiness (r=0.27), and dizziness/shakiness (r=0.20). Must have foods correlated with worrying (r=0.19), lack of sex drive (r=0.20), and food cravings (r=0.22). Having a waistline larger than hips correlated with age (r=0.23), waist circumference (r=0.28), lack of sex drive (r=0.21), and leg cramps/blurred vision (r=0.23). Gaining weight after age 30 correlated with glucose AUMC (r=0.22), age (r=0.64), DBP (r=0.24), waist circumference (r=0.37), BMI (r=0.21), fat mass (r=0.25), body fat % (r=0.36), insomnia (r=0.21), lack of sex drive (r=0.32), being more than 25 lbs overweight (r=0.20), and being overweight since youth (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)
Maternal high fat diet during pregnancy induces similar miRNA changes in F1 generation male germ cells and F3 generation female mammary glands (LB309)
Authors: Nguyen Nguyen , De Assis Sonia , Yin Chao , Wehrenberg Bryan , Hilakivi‐Clarke Leena
Published: June 2021

Maternal dietary exposures during pregnancy can initiate epigenetic inheritance of increased breast cancer risk, perhaps including changes in miRNAs. In this study, we investigated whether the epigenetic inheritance occurs through paternal lineage by studying changes in miRNAs in the male germ cells. Pregnant C57BL/6NTac mothers were fed either a control (CON, 6% corn oil) or isocaloric high n‐6 polyunsaturated fat (HF) diet (18% corn oil) between gestation days 8 and 21. Offspring in the subsequent generations (F1‐F3) were not exposed to any further dietary modifications and were kept on CON diet. F3 female HF offspring exhibited significantly higher mammary tumor incidence (50% had mammary tumors) than the control offspring (32%). We extracted germ line RNA from the sperm cells of 50‐day‐old F1 male offspring and RNA from the mammary glands of F3 female offspring. Expression levels of miRNAs were examined between CON and HF sperm cells and between CON and HF exposed mammary glands. A total of 10 miRNAs were significantly different (p<0.05) in the germ cells of F1 male offspring and 25 miRNAs were different in the mammary glands of F3 females. In addition, four miRNAs were down‐regulated in both F1 male germ cells and F3 mammary glands in the HF offspring: miR‐451, miR‐486, miR‐877, and snoRNA‐202. Down‐regulation of miR‐451 and miR‐486 have been linked to increased susceptibility to many cancers, including breast cancer. Further, recent findings suggest that snoRNAs are new players in cancer. Our study shows that miRNA modifications in F1 generation male germ cells, caused by in utero exposure to a high fat diet, might be carried through to their granddaughters in the F3 generation.

Grant Funding Source: Supported by NCI RO1
Hyperlipidemia induces lipid accumulation and gene expression in adipocytes (LB317)
Authors: Li Yi , Conzalez Trevor , Fernandez‐Alegre Claudia
Published: June 2021

Obesity is becoming a more and more serious health issue in the US and worldwide. This situation only started since a few of decades ago. It is certain that the human genomic coding has not been changed so dramatically by evolution during such short period of time. Evidence indicates that environmental factors including diets and behaviors are involved in affecting gene expressing in developing of human diseases. Since adipose tissues are significantly expanded in individuals with obesity, adipocytes were cultured in the conditions to mimic hyperlipidemia conditions that happen during the developmental process of obesity. We have observed that saturated lipid at high concentrations induces lipid accumulation in adipocytes. A few of new candidate genes that are regulated by the mimic hyperlipidemia conditions have been identified. These genes will be validated by qRT‐PCR and studied further.

Grant Funding Source: Suported by startup funds from Texas A&M University‐Kingsville to YL
The effects of a multi‐herbal mixture HT042 on height gain in short children: a double‐blind, randomized, placebo‐controlled trial (LB322)
Authors: Song Jungbin , Lee Donghun , Kim Hyun Soo , Lee Hyun Jeong , Park Juyeon , Kim Hocheol
Published: June 2021

HT042 is a multi‐herbal mixture consisting of Eleutherococcus senticosus, Astragalus membranaceus and Phlomis umbrosa which have been used to treat children with developmental delay in traditional Asian medicine. A double‐blinded, randomized, placebo‐controlled study was conducted to investigate efficacy of HT042 for short children. We studied 99 children aged from 7 to 12 years, all of whom had an initial height below the 25th percentile for age and sex. The children were randomly assigned to receive 750 mg of HT042 or placebo twice daily for 12 weeks. Anthropometric data including weight, height, height percentile, height standard deviation scores (SDS), fat mass and fat free mass were collected at baseline and at week 4, 8, and 12. Serum levels of growth hormone (GH), insulin‐like growth factor (IGF)‐1 and insulin‐like growth factor binding protein (IGFBP)‐3; nutrient intakes and the amount of time spent exercising were measured at baseline and at week 12. Mean change in height from baseline to week 12 for children who received HT042 and placebo were 2.2 ± 0.7 and 1.9 ± 0.7 cm, respectively (p=0.047). Children in HT042 group showed significant increase in height percentile and height SDS from baseline to week 12 (both p< 0.001). Treatment with HT042 significantly increased fat‐free mass and decreased fat mass from baseline to week 12 (p< 0.001 and p=0.023, respectively) while treatment with placebo did not significantly change both parameters. There was a significant difference in the level of IGFBP‐3, but not GH or IGF‐1, between placebo and HT042 group at week 12 (3386.7 ± 737.0 ng/ml vs. 3064.7 ± 723.6 ng/ml, p=0.039). No significant difference was observed between two groups in nutrient intakes or the amount of time spent exercising at the end of the study. HT042 treatment for short children can increase height and produce positive changes in body composition, suggesting that HT042 would be an effective dietary supplement for increasing height of children.
Increased body mass index adversely affects dyslipidemias, blood pressure and inflammation in men and women with metabolic syndrome (LB365)
Authors: Andersen Catherine , Aguilar David , Jones Jennifer , Blesso Christopher , Al‐Sarraj Taif , Dugan Chrsitine , Barona Jacqueline , Fernandez Maria‐Luz
Published: June 2021

We investigated the effects of body mass index (BMI) on the parameters of metabolic syndrome (MetS) in men (n = 97) and women (n = 165) with MetS. MetS parameters were determined using the National Cholesterol Education Program: Adult Treatment Panel III (NCEP: ATP III) criteria. We classified subjects into four BMI subgroups: normal MetS (BMI < 24.9 kg/m2, n= 11); overweight MetS (BMI 26‐29.9 kg/m2 n =57); obese MetS (30‐39.9 kg/m2, n =157) and morbidly obese MetS (BMI > 40 kg/m2, n = 35). In addition to the expected significant differences between groups in body weight and waist circumference (P < 0.001), there were significant differences in plasma lipids, blood pressure, plasma insulin and C‐reactive protein (CRP) associated with BMI classification.

HDL‐cholesterol was significantly lower in the morbidly obese MetS group (43 ± 9 mg/dL) when compared to MetS subjects with a normal BMI (51 ± 11 mg/dL). In contrast, LDL‐cholesterol was incrementally higher as BMI increased (normal MetS: 111.7 ± 16.9 mg/dL vs. morbidly obese MetS: 129.0 ± 12.5 mg/dL).

Similarly both diastolic and systolic blood pressures were lowest in the normal BMI group and higher in the other BMI groups. Plasma insulin and CRP were also significantly increased from the lowest to the highest BMI (P < 0.05). These findings suggest that even in those with MetS, as BMI increases, there is a higher risk of dyslipidemias, hypertension and inflammation

Grant Funding Source: Egg Nutrition Center and Dairy Council
Sex determination from the distal humerus of 3D model based on the morphometric assessment (LB38)
Authors: Jung Go‐Un , Kim Dong‐Ho , Kwak Dai‐Soon , Lee U‐Young , Kim Myeoung Ju , Kim Yi‐Suk
Published: June 2021

As for sexual dimorphism, the carrying angle has reported focusing on its evident disparity between sexes, so the distal humerus contributing to the formation of elbow joint is anticipated to be useful bone element for estimating sex in forensic anthropology. The purpose of this study is to develop sex‐determining equations from the distal humerus based on the 3D morphometrics and investigate whether the metric method for sexing the distal humerus could be helpful in diverse fields. The right 3D humeral models were obtained from 72 subjects (36 males and 36 females) from the CT‐based Digital Korean Collection. Seven variables including subtrochlear angle (SA), trochlear asymmetry (TA), angle of trochlear inclination (ATI), index (IOF) and depth (DOF) of olecranon fossa, dorsal projection of medial epicondyle (DPME), and distal humeral dimension (DHD) were measured. DOF, DHD, and DPME were selected for providing the equation and the prediction accuracy of 93.1% was postulated through the discriminant function analysis (Discriminant Function = 1.038 x DHD + 0.399 x DPME ‐ 0.383 x DOF). In conclusion, the equation prepared in this study would be expected as a useful method for sex determination especially when the bones are found as fragmentary conditions at excavation site.

Grant Funding Source: This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education (NRF‐2013R1A1A2009306) and also by the NRF‐Global Ph.D. Felloship Program (NRF‐2013H1A2A1033069).
Detection of cranberry and blueberry (Vaccinium sp.) DNA by PCR amplification of the MatK gene. (LB386)
Authors: Herbst Nash , Wilson Ted , Klein Jennifer , Cooper Scott
Published: June 2021

Potential liability related to food allergy and adulteration make documentation of food purity valuable for health and industry. MatK is a chloroplast gene in all plant species that can be used to identify a species based on the presence of an observed DNA sequence. This study sought to develop MatK primers and a PCR method for detection of cranberry (Vaccinium macrocarpon) DNA in cranberry food products. Genus specific MatK PCR primers were designed from multiple sequence alignments. DNA was isolated from homogenized whole fruit, juice, and solid foods followed by precipitation with isopropanol. DNA was amplified by PCR and detected with Sytox Green assays and gel electrophoresis. The MatK primers demonstrated specificity within the genus Vaccinium, but could not discriminate between V. macropcarpon and blueberry (V. corymbosum) after an optimized annealing temperature for PCR was established.

The assay could discriminate Vaccinium DNA from that of other fruits including grape, apple and pear. DNA was isolated from bottled cranberry juice and sweetened dried cranberries, but shearing of the cranberry DNA during processing may have been responsible for a lack of observed PCR product. We have developed primers that can detect DNA from cranberry and blueberry. Future work will redesign the primers to make them species specific DNA for cranberry fruit and processed cranberry food products.
Understanding the effects of herbal bael fruit juice on some medications (LB391)
Authors: Warisnoicharoen Warangkana , Panyapojsak Duangjai , Niwattisaiwong Nuansri , Lawanprasert Somsong
Published: June 2021

The information about interaction between food and medications is still useful for the consumers. A bael fruit juice is the water extract from plant Aegle marmelos Correa. Due to a very pleasant flavor of the fruit juice, it is normally used as a refreshing drink. Moreover, a bael fruit juice is recommended as herbal remedy for hyperglycemia, fever, dysentery and diarrhea. The aim of this study was to investigate the change in the activity of cytochrome P450 3A4 (CYP3A4) as a major drug metabolizing enzyme in human by the fruit juice. A level of furanocoumarins in a fruit juice was quantitatively determined by a reversed‐phase high performance liquid chromatography. A relationship between furanocoumarins and the enzymatic activity might provide understanding on the effect of the juice with medications metabolized by CYP3A4. From the results, the bael fruit juice scarcely decreased the activity of CYP3A4. Either freshly made juice or commercially processed juice contained a trace of furanocoumarins. The findings suggest that the bael fruit juice should not cause interaction with co‐administrative medications metabolized by CYP3A4. It was probably owing to very low amount of furanocoumarins in the juice and other constituted chemicals in the juice have no effects on activity of the enzyme.
β‐carotene inhibits neuroblastoma tumorigenesis by regulating cell differentiation and cancer stem cell stemness (LB400)
Authors: Lim Ji Ye , Kim Yoo‐Sun , Kim Kyung‐Mi , Min Soo‐Jin , Kim Yuri
Published: June 2021

Neuroblastoma (NB) is the most common extracranial solid cancer in young children and malignant NB cells possess cancer stem cells (CSCs) characteristics. The successful elimination of CSCs is a good strategy for developing effective preventive and chemotherapeutic agent. CSCs possess characteristics, including differentiation and turmorigenicity. Although β‐carotene has been associated with many anticancer mechanisms, there is the lack of evidence in the efficacy β‐carotene on CSCs. In the present study, we investigated the effects of β‐carotene on tumor cell differentiation and tumorigenicity using xenograft model which were injected with SK‐N‐BE(2)C NB cells subcutaneously. We found that β‐carotene supplementation significantly inhibited tumor incidence as well as growth compared to tumor control. Furthermore, β‐carotene was shown to induce tumor cell differentiation by up‐regulating vimentin, peripherin, and neurofilament expression. On the other hand, β‐carotene significantly suppressed tumor stemness by down‐regulating CSCs markers such as Oct3/4 and DLK1 expression. It has been reported that the increased expression of hypoxia‐inducible factor1‐α (HIF1‐α) is associated with cancer stemness. Our result has shown that β‐carotene treatment significantly down‐regulated HIF1‐α expression and its downstream target, vascular endothelial growth factor (VEGF). Taken together, these results suggest β‐carotene has chemotherapeutic effects on NB cells by regulating cell differentiation and CSCs stemness.

Grant Funding Source: This research was supported by the National Research Foundation of Korea (Project No. 2011‐0008169) and Brain Korea 21 plus project (Project No. 22A20130012143).
Optimizing periconceptional folic acid supplementation: Steady‐state folate pharmacokinetics in pregnancy (LB416)
Authors: Shere Mahvash , Nguyen Patricia , Tam Carolyn , Stern Seth , Kapur Bhushan , O'Connor Deborah , Koren Gideon
Published: June 2021

Background: Folic acid supplementation before and during pregnancy reduces the risk of neural tube defects. Although previous studies have investigated folate status in non‐pregnant women of childbearing age, steady‐state folate pharmacokinetics have never been evaluated in pregnant women to inform healthcare recommendations regarding the timing of prenatal multivitamin supplementation.

Objective: To compare the steady‐state periconceptional and gestational RBC and plasma folate levels in pregnant women who supplement daily with prenatal multivitamins containing 1.1mg (regular dose) vs. 5mg (high dose) folic acid.

Methods: 37 women, between 18‐45 years of age, who were early in pregnancy or trying to conceive, and were not previously taking folic acid‐containing supplements, were enrolled in this open‐label, 2‐arm, randomized clinical trial after obtaining informed consent. Participants were randomly assigned to take either 1.1mg or 5mg of folic acid‐containing prenatal multivitamins daily till 30 weeks gestational age (g.a.). Plasma and RBC folate levels were measured at baseline, and at 6, 12 and 30 weeks of gestation using a competitive‐binding receptor assay.

Results: RBC folate levels significantly increased in both groups from baseline during pregnancy (p<0.0005). Sustained significant increase in RBC folate was observed in the 5mg group over the course of pregnancy between 6 and 30 weeks gestation, and between 12 and 30 weeks gestation (p<0.0001 and p<0.001), whereas a significant increase in RBC folate concentrations was observed in the 1.1mg group only between g.a.12 to g.a.30 (p<0.05). Plasma folate increased in both groups from baseline to 6 weeks gestation, and then decreased over the course of pregnancy between 6 and 30 weeks gestation, but the decrease was not statistically significant. Plasma levels at 30 weeks gestation in both groups were comparable to their respective baseline levels.

Conclusion: The pharmacokinetics of folate in pregnancy are different than among healthy, non‐pregnant adult women. Despite supplementation over an extended period of time, steady‐state does not seem to be achieved in either group. The decrease in plasma folate in pregnancy may have clinical relevance for women with poor folate status, and may reflect the increased folate demands during pregnancy.

Grant Funding Source: Duchesnay Inc.
hNAG‐1 increases survival by regulating metabolism and insulin/IGF‐1/mTOR signaling (LB432)
Authors: Wang Xingya , Kali Chrysovergis , Justin Kosak , Kissling Grace , Li ruifang , Eling Thomas
Published: June 2021

Metabolic dysfunction is a hallmark of aging. High fat diet (HFD) promotes mortality and decreases lifespan in laboratory animals. Nonsteroidal anti‐inflammatory drug‐activated gene‐1 (NAG‐1) has been shown to play a role in adiposity homeostasis and metabolism. However, it has not been determined if hNAG‐1 increases survival and longevity upon high fat diet. Here we show that hNAG‐1 mice have both significantly increased mean and median life spans than wild‐type (Wt) littermates on a high fat diet. Over the course of the study, hNAG‐1 mice displayed reduced body and adipose tissue weight, lower serum IGF‐1, insulin and glucose levels, improved insulin sensitivity, and increased metabolism. Gene expression analysis revealed significant differences in conserved genes that are important regulators of longevity. Phosphorylation of major components of IGF‐1/mTOR signaling was significantly lower in hNAG‐1mice. Our study shows that hNAG‐1 improves mammalian survival by increasing oxidative metabolism and reducing IGF‐1/mTOR signaling. Our study suggest that hNAG‐1 is an important regulator of mammalian longevity and may act as a survival factor with potential therapeutic uses in obesity‐related diseases where life span is frequently shorter.

Grant Funding Source: NIEHS Intramural Research Program (Eling) Z01‐ ES010016‐14
Effects of a high‐protein low‐calorie intermittent‐fast diet on plasma toxins and oxidative stress following weight loss (LB435)
Authors: Gumpricht Eric , Ward Emery , He Feng , Schuler Bradley , Cho Suk , Arciero Paul
Published: June 2021

In obese subjects, the decrease in fat mass with weight loss is generally regarded as beneficial and may favorably alter oxidative stress. However, it also increases the release of polychlorinated biphenyls (PCBs) from fat depots. Thus, the purpose of this study was to examine the effects of a 10 week higher protein‐low calorie diet incorporating intermittent fasting (HPLC‐IF) on fat mass, plasma toxins (PCBs) and oxidative stress biomarkers, thiobarbituric acid reactive substances (TBARS) and total antioxidant capacity (TAC) in 43 obese men (n=22) and women (n=21). Fat mass, plasma PCBs and oxidative stress biomarkers were assessed at the beginning (pre) and end (post) of the intervention. Following the 10 week HPLC‐IF diet, fat mass decreased significantly (P<0.01) in both groups (males, n=21: 47.5 ± 14.8 vs 37.4 ± 13.3; females, n=19: 49.4 ± 9.6 vs 40.8 ± 9, kg) with males losing more fat mass (‐21.8 ± 5.5% vs ‐17.7± 5.3%, P<0.05; respectively). Eight of 11 individual PCB congeners and total PCB concentration increased significantly overall (Pre: 86.7 ± 45.6 vs Post: 115.6 ± 65.9 ng/g lipid; P<0.01). There were no gender‐specific differences in either individual PCB congeners (with the exception of one) or total PCBs. In addition, TAC increased (Pre: 19.0 ± 2.6 vs. Post: 19.9±2.6 nmol/mL; P<0.05), and TBARS decreased (Pre: 0.24 ± 0.16 vs. Post: 0.18±0.11 μM; P=0.01) overall, with no gender‐specific differences. Interestingly, the percent increase in TAC and total PCB concentration were significantly correlated with the decrease in total fat mass (r=‐0.35, P<0.05; r=‐0.42, P<0.01, respectively). These results suggest that an HPLC‐IF diet induces significant reductions in fat mass resulting in elevated PCB concentrations. Nonetheless, this fat loss‐induced increase in circulating toxins occurred in the presence of enhanced antioxidant status. Although additional study is required to determine the relevance of PCB release during weight/fat loss, our results suggest that an HPLC‐IF diet results in significant improvements in both body composition and antioxidant status.

Grant Funding Source: Isagenix International, LLC
Alcohol metabolism are improved by yeast extract mixtures than Hovenia Dulcis mixtures in healthy men: a double‐blind, randomized crossover, placebo‐controlled trial (LB441)
Authors: Moon Min Sun , Cho Bo Ram , Joo Kyung Ho , Nam Choong Woo , Kim Kwang Soo , Choung Se Young
Published: June 2021

Alcohol hangover is an unpleasant aftermath of heavy alcohol drinking. Hangover onset starts after the oral administration of alcohol since ethanol, a key component of the alcoholic drinks, metabolized to acetaldehyde. In this study, we investigated whether the supplement formula might improve the alcohol metabolism in healthy adult men. In a double‐blinded, randomized, crossover study, subjects were received yeast extract mixtures (YEM, n=21), Hovenia Dulcis mixtures (HDM, n=21), placebo (PLC, n=21), and control (CON, n=21) in separate 4 week supplementation periods at a single oral dose followed by one week washout periods. At each visit (0, 1, 2, 3, 4 weeks), the subjects drank 450 mL 23 % alcohol simultaneously with mixtures and blood was drawn periodically (0, 0.25, 0.5, 1, 2, 4, 6, 15 hr). Fifteen subjects completed the protocol and included in the analysis. The plasma ethanol concentrations were tended to lower in the YEM (10 %) and the HDM (5 %) groups. The area under the curves (AUCs) and Cmax for plasma ethanol were significantly decreased only in the YEM group, when compared with the CON group. The AUCs and Cmax for plasma acetaldehyde concentration were significantly decreased in the YEM (45 % and 54 %) and the HDM (35 % and 53 %) groups respectively, when compared with PLC (P<0.01). These findings demonstrated that alcohol metabolism and hangover syndromes might improve by YEM than HDM.
Assessing diet and physical activity related to obesity risk among Ugandan school children (LB462)
Authors: Christoph Mary , Grigsby‐Toussaint Diana , Baingana Rhona , Ntambi James
Published: June 2021

In Uganda, 10.4% and 3.2% of adolescent girls and boys are considered overweight, even in the midst of malnutrition. However, few studies have explored risk factors associated with obesity among adolescents. We conducted a cross‐sectional survey of 151 randomly selected children aged 11‐ 16 years in two central Ugandan rural and urban schools in November 2013. Sociodemographic characteristics, diet, and physical activity were surveyed; height, weight, and waist circumference were measured (kg/m2, cm). Mean age of participants was 12.1 years (±1), with a mean BMI percentile of 43% (49% male). Sixty‐two percent of the children owned TVs, primarily driven by the urban sample, 73% of whom owned TVs. Soda consumption (64% having consumed within the past week) did not differ by school. Underweight was more common than overweight, with 10% of children being below the 10th percentile, and only 5.3% above the 80th percentile. Meat and fruit (but not vegetable) consumption and physical activity differed by rural‐urban residence (P<0.01), with rural children reporting lower physical activity and meat consumption, but higher fruit intake. This study is one of the first to assess the impact of diet and activity among rural and urban school children in Uganda. It provides a basis for understanding the risk factors associated with overweight and obesity in adolescents in Uganda.

Grant Funding Source: This was was supported by the Agriculture and Food Research Initiative of the USDA National Institute of Food and Agriculture as part of the AFRI Childhood Obesity Prevention Challenge (2011‐67001‐30101) to the UIUC Division of Nutritional Sciences.
n‐3 PUFA habitual intake is not associated with immunological biomarkers in Mexican Type 2 diabetes population. (LB466)
Authors: Martinez‐Carrillo Beatriz , Guadarrama‐López Ana , VALDÉS‐RAMOS ROXANA
Published: June 2021

Background: Changes in lifestyle have led to an increase in the global incidence of type 2 diabetes mellitus (T2DM). In Mexico there are 6.4 million type two diabetic patients. It is widely recognized that insulin resistance and T2DM are associated with low‐grade chronic inflammation. There is evidence about the beneficial effect of polyunsaturated fatty acids (PUFA) n‐3 in patients with chronic inflammatory diseases. Objective: To investigate the relationship between habitual fatty acid intake and circulating biomarkers of inflammation in patients with T2DM in the population of the State of Mexico. Methods: We conducted a cross sectional study in a sample of 120 diabetic adults to whom we performed anthropometric assessments (BMI, waist circumference, body fat percentage and blood pressure) as well as PUFA dietary intake, biochemical analyses and inflammation biomarkers (IFN‐γ, TNF‐ α, IL‐1 β). Results: A low intake of n‐3 was found in our population (0.68 ± 0.55 g/day). Moreover higher serum concentrations of pro‐inflammatory cytokines were present. No correlations between the regular consumption of fatty acids and lipid profile and biomarkers of inflammation were found. Conclusions: Mexican diabetic patients had a low PUFA intake, but there is no association between the consumption of PUFA and lipid profile or pro‐inflammatory or anti‐inflammatory biomarkers.
Macrophage migration inhibitory factor has a role controlling colorectal cancer (LB491)
Authors: Rodriguez‐Sosa Miriam , Pacheco‐Fernández Thalia , Juárez‐Avelar Imelda , Rodríguez‐Monroy Marco , Nieto‐Yañez Oscar , Chirino Yolanda , León‐Cabrera Sonia , Pérez‐Plasencia Carlos , Avila‐Moreno Federico , Terrazas Luis
Published: June 2021

Chronic inflammation has been related with cancer development by counteracting and unbalancing the immune response. The inflammatory cytokine macrophage migration inhibitory factor (MIF) is overexpressed in many carcinogenic tumors, stimulates angiogenesis and metastasis promoting proliferation and survival in malignant cells. In the inflammatory disease ulcerative colitis MIF is early expressed and favors the inflammatory cytokine production like TNF‐α, and IL‐1β. As intestinal chronic inflammation may proceed to colorectal cancer, we are interested in determining the role of MIF on development and pathology on colorectal cancer. To elucidate this point we used MIF‐KO and WT mice, both with BALB/c genetic background, to develop a colorectal cancer model of Dextran Sodium Sulfate and Azoxymethane. As expected, MIF‐KO mice showed low levels of serum inflammatory cytokines TNF‐α, IL‐1β and IFN‐γ compared to WT mice. Strikingly, we found significantly more tumor and in a higher development stage on MIF‐KO mice (24.4+/‐2) compared to WT mice (12.6+/‐2). Higher number of activated macrophages and NK cells were detected in intestinal epithelium on WT mice compared to MIF‐KO mice. Therefore, although MIF has been recognized as a promoter of inflammation, which could favor the development of cancer; our results suggest that MIF is involved in controlling the development of colorectal cancer, possibly by its chemokine properties.

Grant Funding Source: Supported by CONACyT (152224) and PAPCA‐2013. The student TPF holds a scholarship from CONACyt, #20465.
Systemic platelet‐activating factor‐receptor activation augments experimental lung cancer growth and metastasis (LB496)
Authors: Sahu Ravi , Hackler Patrick , Konger Raymond , Travers Jeffrey
Published: June 2021

Pro‐oxidative stressors including cigarette smoke (CS) generate novel lipids with Platelet‐activated Factor‐receptor (PAF‐R) agonistic activity and mediate systemic immunosuppression, one of the most recognized events in promoting carcinogenesis. Our previous studies have demonstrated that PAF‐R agonists mediated systemic immunosuppression augments the growth of experimental murine B16F10 melanoma tumors in a PAF‐R dependent manner. As CS generates PAF‐R agonists, the current studies sought to determine the impact of PAF‐R‐agonists on lung cancer growth and metastasis using an established experimental murine lewis lung carcinoma (LLC) model. We show that systemic PAF‐R agonist, CPAF augments the growth of subcutaneously implanted LLC tumors in a PAF‐R dependent manner as this effect was not observed in PAF‐R deficient (PAFR‐KO) mice. We next determined the ability of CPAF‐mediated PAF‐R dependent enhanced LLC tumor growth on lung metastasis. We observed increase number of lung mets only in CPAF‐treated WT and not in PAF‐R‐KO mice. In order to investigate the role of tumor or host cell PAF‐R in these effect, we first determined the expression of the PAF‐R in LLC tumor cells. Our results demonstrate that LLC tumor cells lack functional PAF‐R expression as measured by qRT‐PCR and functional Ca2+ mobilization assay and compared with arrays of PAF‐R sufficient and deficient cells. In addition, CPAF treatment failed to induce increase proliferation of LLC cells, indicating the role of host rather tumor cell PAF‐R in these events. This finding has clinical implications as it provides insight into a mechanism through which lung cancer cells evade immune surveillance.

Grant Funding Source: NIH R01 HL062996, AICR 09A062, ACSIRG 4185607, Showlater Research Trust Fund 4485602
Investigating the effect of antibiotics on gut microbiota components and subsequent Clostridium difficile infection (LB516)
Authors: Schenck Louis Patrick , Hirota Simon , Armstrong Glen , MacDonald Justin , Beck Paul
Published: June 2021

C. difficile (Cdif) infections (CDI) are a world‐wide epidemic. Recent small clinical trials have generated excitement for the use of fecal microbial transplants as a therapeutic option; however, the exact components of the microbiota needed for protection against CDI have remained elusive. This study assessed the specific intestinal microbiota components associated with prevention or exacerbation of CDI in a mouse model of disease. C57/Bl6 mice from two different vendors (A or B) were exposed to broad‐spectrum antibiotics before oral gavage with Cdif spores. Mice from A and B were co‐housed for 14 days, followed by exposure to antibiotics and development of CDI. Microbiota diversity and composition were analyzed using Illumina MiSeq Next‐Gen Sequencing of 16S rDNA in fecal samples. Mice from different vendors had distinct microbiota profiles both before and after antibiotic exposure. Mice from A developed severe CDI as evidenced by weight loss, histological damage, increased MPO levels and inflammatory cytokines, including KC and IL‐1β. Mice from B were resistant to CDI with minimal histological and cytokine changes from baseline. While similar at naïve levels, vendor B retained a high abundance of Bacteroidetes phyla after antibiotics. Following cohousing, mice from vendor A had less severe CDI and mice from vendor B had more severe CDI. Our data are the first to identify specific patterns in the intestinal microbiota that confer susceptibility to CDI. Furthermore, the bacterial populations that may be critical for preventing or reducing the severity of CDI were revealed. This study may lead to more targeted bacteriotherapy for the treatment and prevention of CDI.

Grant Funding Source: Supported by Canadian Institute of Health Research and Alberta Innovates ‐ Health Solutions
Impaired microvascular function and increased vascular endothelial cell angiopoeitin‐2 and tie‐2 mRNA expression in human sepsis (LB527)
Authors: Wegman‐Points Lauren , Lane Abbi , Doerschug Kevin , Delsing Angela , Pierce Gary
Published: June 2021

Sepsis is a systemic inflammatory syndrome resulting in acute organ failure attributable to heterogeneous tissue perfusion and microvascular endothelial vasodilatory and barrier dysfunction. Angiopoeitin‐2 (Ang2), a growth factor released into circulation that binds to the Tie‐2 receptor during sepsis, is a robust predictor of clinical disease severity. However, there is currently no direct evidence in humans that Ang2 and Tie2 mRNA/protein expression are elevated in the endothelium of critically ill patients with sepsis. Microvascular function (finger peripheral artery tonometry) was reduced 44% in septic patients (n=4) vs. age‐matched (n=8) controls (1.480.14 vs. 2.630.17%, P<0.01). Interestingly, Ang2 (+1.7 fold P<0.03) and Tie2 (+2.7 fold, P<0.04) mRNA (qRT‐PCR) were higher (vs. controls), in human aortic endothelial cells incubated 24 hrs in vitro with media + 10% plasma from sepsis (n=6) vs. control (n=6) humans. In a subset, there was no difference in Tie‐2 (n=3, 6.4e5 ± 1.5e5 vs. n=4, 4.4e5 ± 7.4e4, P=0.23) protein expression (corrected fluorescence) in isolated endothelial cells (J wire sampling/immunofluorescence), but Ang2 tended to be lower (n=2, 1.6e5 ± 7.8e3 vs. n=4, 2.1e5 ± 1.6e4, P=0.10) in sepsis vs controls. These data suggest that Ang2 and Tie‐2 gene expression are upregulated in the septic endothelium and lower protein Ang2 expression may indicate extracellular Ang2 protein release.

Grant Funding Source: Supported by NIH 5R01HL092056‐05
Molecular and cellular mechanism of systemic vaso‐occlusion in Sickle Cell Disease (LB544)
Authors: Jimenez Maritza , Sundd Prithu
Published: June 2021

SCD is a genetic disease caused by a single point mutation in the β‐globin gene of hemoglobin, which leads to sickling of RBCs under deoxygenated condition. Sickle RBCs (sRBCs) are not only less deformable, but also express enhanced levels of major adhesion molecules compared to normal discoid shaped RBCs of healthy patients. In addition to RBC sickling, SCD is also associated with a hyper‐inflammatory state, which accounts for enhanced adherence of leukocytes to vascular endothelium. Leukocyte‐endothelium adhesion starts with leukocyte rolling mediated by P‐selectin‐glycoprotein‐ligand (PSGL)‐1 on leukocytes binding to P‐selectin on endothelium. Rolling is followed by firm arrest, which is medicated by activated β¬2‐integrins (LFA‐1 and Mac‐1) on the leukocytes binding to inter‐cellular‐adhesion‐molecule (ICAM)‐1 on endothelium. Although neutrophils have been shown to play a role in the onset of vaso‐occlusion by interacting with sRBCs in systemic venules of SCD mice, the cellular and molecular mechanism that enables systemic vaso‐occlusion in human patients is not completely understood. Heparinized blood from SCD patients and race matched control subjects is perfused through a microfluidic flow chamber with a glass bottom coated with either human microvascular endothelial cells or a cocktail of P‐selection, ICAM‐1 and CXCL‐8 at a physiological shear stress (2‐10 dyn cm‐2). Fluorescent Abs against CD16 and glycophorin‐A are added to the blood to stain neutrophils and sRBCs, respectively and cellular interactions recorded using multi‐color Quantitative Dynamic Footprinting or epifluorescence microscopy. Preliminary results reveal that neutrophil arrest is facilitated by the formation of a thick trunk like structure at the rear, which also enables capturing of free flowing sRBCs by arrested neutrophils. Understanding the molecular mechanism behind vaso‐occlusion will be useful in designing prophylactic interventions for SCD patients.

Grant Funding Source: This study is supported by 11SDG7340005 from the American Heart Association (P.S.), VMI start‐up funds (P.S.) and CBTP‐T32 fellowship (M.J).
Hyperoxia and Regulatory T cell (Treg) development in neonatal mice Hardik Patel1, Angela Leon Hernandez1, Mark Pezzano2 and Mohamed Ahmed1 1Neonatology Research Lab, The Feinstein Institute for Medical Research, Manhasset, NY. 2Department of Biology, The City College of NY, New York, NY (LB546)
Authors: Patel Hardik , Leon Hernandez Angela , Pezzano Mark , Ahmed Mohamed
Published: June 2021

Oxygen is the most common therapy used in Neonatal Intensive Care Unit (NICU). The potential effect of hyperoxic exposure has become a focus of concern. In our preliminary findings, we found decreased levels of CD4+ cells and Tregs (FOXP3+) in the thymus of hyperoxia exposed neonatal mice. Our aim is to find the mechanism by which hyperoxia decreases Treg cells in neonatal mice thymus. Chemokines such as CCL19 and CCL21 and their receptors CCR7, CCR9 and CxCR4 are crucial for development of Treg cell in the thymus. Therefore, we hypothesized that hyperoxia disrupt Treg cell development through an imbalance in chemokine expression or its target receptors.

Neonatal mice were housed either at room air (FiO2 21%), or hyperoxia (FiO2 95%) for five days. After exposure, thymus immunostainined for FOXP3, CD4, CCL19 and K14. Chemokines CCL19 & 21 expression was done using RT‐qPCR. CCR7, CCR9, CxCR4 and FOXp3 were assayed using for flow cytometery.

Immunostaining showed arrest of FOXP3 positive cells at the cortico‐medullary junction and increased expression of CCL19 in cortical area of thymus of hyperoxia group. CCL19 and CCL21 mRNA levels were higher in hyperoxia group. CCR7, CCR9 and CxCR4 chemokine receptors were significant decrease on double negative (CD4‐CD8‐) cells. CCR7 expression was also significantly decreased on FOXP3 expressing double positive cells.

Suppression effect of hyperoxia on Treg development and maturation is possibly due to down regulation of receptor (CCR7, CCR9 and CXCR4) expression in the early developmental stage. Treg cells arrest in the cortico‐medullary junction, even with higher expression of chemkines, is possibly related to low CCR7 expression on CD4+ CD8+ double positive cells.
Discovery of thienoquinolin urea transporter inhibitors acts as a potential diuretic (LB587)
Authors: Li Fei , Yang Baoxue
Published: June 2021

The motivation for this study was the need for novel diuretics in order to circumvent the side effects associated with long‐term use of conventional diuretics. Urea transporters (UTs) are a family of membrane channel proteins that are specifically permeable to urea and play an important role in intrarenal urea recycling and in urine concentration. Using an erythrocyte osmotic lysis assay, we screened a small‐molecule library for inhibitors of urea transport. A novel class of thienoquinolin UT‐B inhibitors were identified, of which PU‐14 had potent inhibition activity on human, rabbit, rat, and mouse UT‐B. The half‐maximal inhibitory concentration of PU‐14 on rat UT‐B‐mediated urea transport was ~0.8 μmol/l, and it did not affect urea transport in mouse blood cell lacking UT‐B but inhibited UT‐A‐type urea transporters. PU‐14 showed no significant cellular toxicity at concentrations up to its solubility limit of 80 μmol/l. Subcutaneous delivery of PU‐14 (at 12.5, 50, and 100 mg/kg) to rats caused an increase of urine output and a decrease of the urine urea concentration and subsequent osmolality without electrolyte disturbances and liver or renal damages. This suggests that PU‐14 has a diuretic effect by urea‐selective diuresis. Thus, PU‐14 or its analogs might be developed as a new diuretic in diseases associated with water retention without causing electrolyte imbalance. PU‐14 may establish ‘chemical knockout’ animal models to study the physiological functions of UTs. Further studies will be focused on the structural modification and characterization of pharmacokinetics and the absorption, distribution, metabolism, excretion, and toxicity properties of PU‐14.
Teriflunomide is a specific and clinically relevant BCRP probe at the blood‐brain barrier (LB591)
Authors: Nagy Ildikó , Beéry Erzsébet , Erdő Franciska , Sziráki István , Sike Mirabella , Molnár Éva , Krajcsi Péter
Published: June 2021

Background: Disease modifying antirheumatic drug (DMARD’s), leflunomide, is a non‐cytotoxic proliferation inhibitor of mitogen‐stimulated T and B lymphocytes. Leflunomide is a prodrug that undergoes rapid conversion to its active form, teriflunomide.The latter is poorly metabolized and is eliminated mainly through direct biliary excretion of unchanged drug as well as renal excretion of metabolites. It can take several weeks or months for DMARDs to display a clinical effect which often leads to loss of efficacy in patients over time. Resistance to DMARDs may be a multifactorial phenomenon including enhanced drug efflux by ABC transporters. Recently it was reported that leflunomide and its metabolite, teriflunomide, are high‐affinity substrates of BCRP transporter in vitro. Aim: Assess whether teriflunomide can used as a specific and clinically relevant BCRP probe at the blood‐brain barrier, using in vitro and in vivo models. Methods and results: Efflux assays in Caco2 and MDCKII monolayers overexpressing BCRP were used to determine whether teriflunomide transport is BCRP specific in‐vitro. In these systems teriflunomide was successfully transported and could be inhibited by BCRP specific inhibitor Ko143. Subsequently, brain penetration of teriflunomide was tested in anesthetized mice in dual‐probe microdialysis experiments. Comparing teriflunomide levels in dialysate samples from treated and control animals, administration of Ko143 resulted in an increased brain penetration of teriflunomide. Conclusion: Taken together our results are well correlated, and confirmed that BCRP plays an important role in the pharmacokinetics of teriflunomide which meets the criteria for being a clinically relevant BCRP probe.