bluuebunny/crossref_metadata_2025 huggingface dataset:
Sorghum‐based dietary intervention enriches Faecalibacterium prausnitzii in fecal samples of overweight individuals
Authors: Seidel Derek Vaughn , Martínez Ines , Taddeo Stella S , Zoh Rszoh , Haub Mark D , Walter Jens , Turner Nancy D
Published: June 2021

The gut microbiota impacts host metabolism with implications for obesity and related metabolic disorders and affects intestinal inflammation. We have shown that a polyphenol‐enriched sumac‐sorghum diet alters microbiota and short chain fatty acid (SCFA) production in rats. The goal of this study was to determine if similar effects occur in overweight subjects predisposed to metabolic syndrome. Human subjects (n=25) were randomized in a crossover design with high (H) and low (L) intakes of sumacsorghum breakfast cereal incorporated in their normal diets. Fecal samples (pre and post treatment) were used to characterize microbial composition by 454 pyrosequencing of 16S rRNA PCR amplicons and to measure SCFA concentrations. We found that sumac‐sorghum induced compositional changes in the fecal microbiota of the subjects. Most importantly, the antiinflammatory commensal Faecalibacterium prausnitzii, which has been shown to be a major butyrate producer in the human gut, significantly increased in both treatment groups (p=0.0006). In response to the L diet, fecal butyrate tended to increase (+9.9%), whereas with the H diet, butyrate tended to decrease (−12.5%). This study revealed that sumac‐sorghum increased the putatively health promoting bacterium Faecalibacterium prausnitzii, which may inhibit intestinal inflammation in overweight subjects. Funded by United Sorghum Checkoff Program R0002–11.
Weight Control Attitude and Nutrition Knowledge of Korean High School Students by BMI
Authors: Chang Kyung Ja , Cho Ha‐Yun
Published: June 2021

The purpose of this study was to investigate the weight control attitude and nutrition knowledge of Korean high school students for nutrition education related to weight control. The subjects were 479 high school students (male 247, female 232) in Incheon metropolitan city, Korea. A cross‐sectional survey was conducted using a questionnaire from July 27 to 29, 2011. The data were analyzed statistically with SPSS 12.0 program. As for the distribution of weight by BMI, there was a significant difference between male and female students; 50.2% of the male normal, 19.8% overweight, 16.2% obese, 13.8% underweight and 61.6% of the female normal, 20.7% underweight, 9.1% obese, 8.6% overweight. As for satisfaction with body image, the underweight students were significantly more satisfied with their body image, and the male students were significantly more satisfied compared to the female students. Also the obese or female students that were less satisfied with body image had significantly more interest in weight control. The obese students had significantly more experience to lose weight. As for effective ways of reducing weight, 62.6% answered exercise, 34.8% did diet therapy. As for nutrition knowledge scores, the female students (3.46/5) showed significantly higher average score compared to the male students (3.01/5) and there was no significant difference in nutrition knowledge scores according to BMI. Therefore, nutrition education for personalized weight control according to gender and BMI should be provided in high school in Korea.
After‐school obesity prevention programs: Are engaged parents a myth or reality?
Authors: Spence Marsha , McDonald Aneisa , Perry‐Burst Carolyn
Published: June 2021

Evidence suggests that parental involvement in school‐based obesity prevention programs for elementary school students may improve outcomes. However, parental engagement can be difficult to achieve and maintain, especially among low‐income and rural families. As part of a continuous quality improvement process for an afterschool obesity prevention program for 4th and 5th grade students in East Tennessee, 10 parent focus groups (n=4–14 participants/group) were conducted at the conclusion of the programs to ascertain parents’ perspective about best practices to increase parental involvement and preferred program activities to assist in implementing behavioral changes. The focus group recordings were transcribed verbatim, coded, and emergent themes were extracted, using grounded theory methods. Preliminary findings suggest that although parents desire to make healthful changes for their families, conflicting nutrition messages in the media are major barriers to actually implementing changes. Additional themes that emerged include time constraints for attending parental programs and for meal preparation, lack of skills to prepare meals, and parents’ physical limitations that restrict physical activity. These findings provide insight into ways to engage parents and assist them in making nutrition and physical activity changes with their children.
Dentists′ Dietary Perception and Practice Patterns in a Dental Practice‐Based Research Network
Authors: Yokoyama Yoko , Kakudate Naoki , Sumida Futoshi , Matsumoto Yuki , Gilbert Gregg H , Gordan Valeria V
Published: June 2021

Objective

(1) To examine discordance between dentists’ perception of the importance of diet in caries treatment planning and their actual provision of diet counseling to patients, and (2) identify dentists’ characteristics associated with their provision of diet counseling.

Methods

The cross‐sectional study was conducted using US the National Dental Practice‐Based Research Network (PBRN) questionnaire. The study queried dentists working in outpatient dental practices who were affiliated with the Dental PBRN Japan (n=282).

Results

The majority of participants (n=116, 63%) recognized that diet is important to oral health. However, among participants who think diet is important (n=116), only 48% (n=56) provide diet counseling to more than 20% of their patients. Multiple logistic regression analysis suggested that several variables were associated with providing diet counseling: dentist gender, percentage of patients interested in caries prevention, and who receive blood pressure screening.

Conclusions

Some discordance exists between dentists’ perception of the importance of diet in caries treatment planning and their actual practice pattern regarding diet counseling to patients. Reducing this discordance may require additional dentist education, including nutritional and systemic disease concepts; patient education to increase understanding of the importance of caries prevention.

Grant Funding Source: NIH grants U01‐DE‐16746, U01‐DE‐16747, and U19‐DE‐22516
HSCCC isolation and characterization of walnut polyphenols with antioxidative and lipolytic activity
Authors: Grace Mary H , Neff Scott A , Warlick Charles , Ayoub Elaria , Esposito Debora , Wilson Mickey , Komarnytsky Slavko , Lila Mary Ann
Published: June 2021

In order to characterize the structures, biological activity, and mechanisms of action for walnut constituents, the key phytochemicals must be isolated in sufficient quantities. The objective of this research was to establish a method for rapid separation of walnut polyphenols which preserves structural integrity and yields high quantities and purities. A robust HSCCC method was developed to fractionate ethyl acetate and butanol extracts. This method efficiently yielded gallic acid, catechin, ellagic acid, casuarictin, pedunculagin, tellimagrindin I, 2,3‐O‐hexahydroxydiphenoyl‐ glucopyranoside (2,3‐HHDP‐glc), strictinin, and α‐hydrojuglone 4‐glucoside in 1 to 2 runs, and allowed us to test the ability of these walnut components to inhibit LPS‐induced ROS in human neuroblastoma (SH‐SY5Y) cells and reduce lipid accumulation in mouse adipocytes (3T3‐L1). The ethyl acetate and butanol extracts, as well as isolated ellagitannins, decreased ROS generation in SH‐SY5Y cells. Additionally, the 2,3‐HHDP‐glc decreased lipid accumulation in a dose‐dependent manner, resulting in 14% reduction of intracellular fat at 90 μM after 24 h of treatment. Treatment with 2,3‐HHDP‐glc increased basal oxygen consumption rate by 24.9% but had no effect on extracellular acidification rate, suggesting that it decreased lipid content of adipocytes by stimulating mitochondrial respiration. (Support from California Walnut Commission)
Is There Fruit In That? Children Are Misled By Television Food Advertisements
Authors: Heller Rebecca , Berhaupt‐Glickstein Amanda , Martin‐Biggers Jennifer , Byrd‐Bredbenner Carol
Published: June 2021

Fruit‐flavored food advertisements on television often contain verbal (e.g., words like fruit, fruity, juice) and visual (e.g., images, art) references to fruit despite a lack of real fruit in the product. This study determined how information in television food ads affected children's (4–7y; n=49; 65% male) perceptions of foods’ fruit content. Each child was shown the ads (n=16) in random order and, after each, asked, “Did they use fruit to make this?” The ad groupings were for 100% real fruit foods (n=8) and foods containing 0% real fruit with artificial fruit‐flavors as their only fruit “content” (n=8). Each ad grouping had 4 categories with 2 ads/category: neither fruit images nor verbal references, fruit images, verbal fruit references, and both fruit images and verbal references. All ads were for foods not distributed in the study area to control for familiarity. The presence of fruit images in ads led to the perception that the food contained real fruit, regardless of its true fruit content. The accuracy with which they could identify whether a food was made with fruit was significantly lower for ads of foods containing 0% fruit when the ad had a fruit‐related verbal (p=0.017), visual (p<0.0001), or both references (p<0.0001). Results suggest that children are deceived about foods’ real fruit content when ads for artificially fruit‐flavored foods containing 0% real fruit include fruit images.

Grant Funding Source: NJ Agricultural Experiment Station
Protective effects of different dietary proportion of fish oil on hepatic injury in chronic ethanol‐fed rats
Authors: Yang Suh‐Ching , Peng Hsiang‐Chi , Chen Ya‐Ling , Liao Wei‐Hsiang , Wang Xiang‐Dong
Published: June 2021

Excess consumption of alcohol is known to cause liver damage. The present study was to investigate the effects of different proportions of fish oil on hepatic aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities and oxidative stress in ethanol‐fed rats. Male Wistar rats were divided into six groups (n=5/group) and fed with either the control diet or the alcohol diet, in which the fat composition of both diets was adjusted with 25% or 45% fish oil substitution for olive oil. The groups were C (control), 25%CF (control with 25% fish oil), 45%CF (control with 45% fish oil), E (ethanol), 25%EF (ethanol with 25% fish oil), and 45%EF (ethanol with 45% fish oil). Rats were sacrificed after 8 weeks of the treatments. Results showed that plasma levels of AST, ALT activities, 8‐isoprostaglandinF2α, triglyceride and total cholesterol were significantly decreased and hepatic GSH/GSSG ratio was significantly increased in the 25%EF and 45%EF groups than those in the E group (p<0.05). In addition, the pathologic scores for microvesicular fatty change, degeneration and necrosis were significantly decreased in 25% EF and 45% EF groups as compared with that in the E group (p<0.05). The results suggested that fish oil substitution could prevent ethanol‐induced liver damage. Supported by NSC100–2320‐B‐038–023‐MY3.
Association of Lead Exposure and Untargeted Metabolomics with BMI and Hormones in Adolescence
Authors: Peterson Karen E. , Dolinoy D , Burant C , Lee J. , Sanchez B , Zhang Z , Yang T C. , Goodrich J , Ettinger A , Meeker J , Hu H , Solano M , Wang N , Tellez‐Rojo MM
Published: June 2021

Lead, an endocrine disrupting chemical (EDC), has been related to delays in weight and pubertal onset, but limited data account for metabolic mechanisms. We examined the additional variance in Body Mass Index (BMI), estradiol and testosterone among n=40 children aged 7–15 yr predicted by metabolomics, using a multi‐index model with variable selection to construct 2 indices for % methylation of CpG sites in utero and peripuberty of 4 candidate genes associated with physical growth and 2 indices each built from 317 metabolites with known functions and 440 so called “known‐unknown” compounds. After adjusting for child sex, age, leptin, blood lead at 4 yr and % methylation, the R2 for BMI increased from 0.81 to 0.95, when metabolites were included in the model. For estradiol and total testosterone, the R2increased from 0.51 and 0.33, without metabolites, to 0.95 and 0.94 with metabolites, respectively. Using Metscape 2.0 to visualize the pathways associated with lead exposure, we found bile acid, amino acid and nucleotide metabolism were enriched. Results suggest our capacity to predict BMI, and hormonal markers of sexual maturation related to EDCs is markedly improved with inclusion of metabolites. Research support: NIEHS/EPA P20 ES01817101/RD834800; NIEHS R01 ES007821; P30 ES017885; P30 DK 089503.
Fructose intake and circulating triglycerides: an examination of the roles of APOC 3 and FOXO1
Authors: Campbell Eric S , Castonguay Thomas W
Published: June 2021

Fructose consumption can promote hypertriglyceridemia (HTG) in both humans and laboratory animals. We have recently reported that giving rats overnight access to a 16% fructose solution results in HTG. Several investigators have suggested that hepatic FOXO1 and/or APOC 3 may be implicated in promoting fructose‐induced HTG. We have found that neither FOXO1 nor APOC 3 expression in the livers of rats fed fructose differed from controls that were fed lab chow only. Similarly, neither FOXO1 nor APOC 3 expression in rats fed glucose, sucrose or high fructose corn syrup solutions were significantly different from controls. These results suggest that the changes in FOXO1 and APOC 3 following fructose consumption are secondary to the changes necessary to cause fructose‐induced HTG. Alternatively, hepatic FOXO1 and/or APOC 3 expression may have been returned to pre‐fructose levels prior to sampling. In future experiments we intend to monitor postprandial hepatic expression of FOX01 and APOC 3 in hourly intervals to better characterize the onset of HTG. Supported in part by a grant from the Maryland Agricultural Experiment Station.
Rice bran protein concentrate can reduce fat accumulation in rats fed high‐fat diet
Authors: Yu Jaeran , Lee Okhee
Published: June 2021

Rice is a staple food and major source for energy intake in Korean population. Furthermore, it would be the highest protein source in Korean diet, because the white rice is the most consumed food according to 2005 KNHANES. Rice bran, outer shell of the rice milling process has 10~16% highly nutritional protein itself. Rice bran protein has been reported to have anti‐obesitic effect, However the feasibility of rice protein is not well known. Therefore, we examined the effect of rice bran protein on body far accumulation in rats fed high‐fat diet based on a AIN 93M(CO), and high fat diet with rice bran protein 0% (CA), 10%(RB10) 20%(RB20), and with soy isolated protein(SP). After 50 days, the rats were sacrificed. The complete substitution of casein(CA) with rice bran protein(RB20) reduced significantly final weight and epididymal fat pad of rats fed high‐fat diets. The fecal fat excretion was the highest in the RB20 group among experimental groups. Total lipid and triglyceride concentration in liver of RB10 and RB20 groups were significantly lower than those of CA. The concentration of albumin and total protein in plasma and liver did not show any differences by the replacement of casein with rice bran protein. This study suggests that the reduction of fat accumulation can be achieved in rats by replacing casein protein with rice bran protein without reducing visceral protein content.

Grant Funding Source:No
Perturbed 1‐carbon metabolism alters bile acid pools and insulin signaling
Authors: Corbin Karen D , Mei Shuang , Cao Wenhong , Zeisel Steven H
Published: June 2021

Betaine homocysteine methyltransferase (BHMT) is responsible for the utilization of betaine, a metabolite of choline, as a methyl donor. We previously demonstrated that Bhmt knockout (KO) mice display several metabolic changes including alterations to the bile acid pool, insulin sensitivity, increased energy expenditure, hepatic steatosis and reduced fat pads. To begin exploring the signaling pathways integrating the observed phenotypes, we asked whether hepatic insulin signaling is altered in Bhmt KO mice. Our preliminary results demonstrate enhanced insulin signaling as evidenced by increased concentration of insulin receptor and phosphorylation of Akt at serine 473. Given that Bhmt KO mice also have hepatic steatosis, we asked whether the enhanced insulin signaling was accompanied by increased expression of SREBP1c. We found that SREBP1c precursor protein expression was increased. Our findings suggest that 1‐carbon metabolism intersects with glucose and lipid signaling. Ongoing studies are designed to test the hypothesis that the energy metabolism phenotypes associated with disruption of 1‐carbon metabolism are mediated by alterations in bile acid signaling via the farnesoid X receptor.
Assessment of folate, vitamin B6 and vitamin B12 status among newly diagnosed Omani cardiac patients
Authors: Waly Mostafa I. , Ali Amanat , Al‐Farsi Yahya M. , Ouhtit Allal , Al‐Nassri Amira , Arafa Mostafa
Published: June 2021

Background

Oman is currently witnessing an increase in the rate of morbidity and mortality from non‐communicable diseases, including cardiovascular diseases (CVD).

Objectives

This study was aimed to assess the nutritional and biochemical status of folate, vitamin B6 and vitamin B12 in relation to homocysteine level in newly diagnosed Omani cardiac patients.

Methods and Subjects

A case‐control study included 25 cases, recently diagnosed Omani cardiac patients, and 25 age and gender matched healthy subjects as a control group.

Results

As compared to control group, the enrolled Omani cardiac patients had lower dietary intake of folate, vitamin B6 and vitamin B12 that is concomitant with low serum levels of these three nutrients, P<0.05. The patents had hyperhomocysteinemia that was negatively correlated with the status of folate, vitamin B6 and vitamin B12, and they consumed high protein and saturated fats from animal sources in amount that exceeds the recommended daily dietary intake.

Conclusion

Hyperhomocysteinemia and low status of folate, vitamin B6 and vitamin B12 are involved in the aetiology of CVD among high risk groups.
Breastfeeding and Birth Order Modify Food Insecurity's Associations with Inadequate and Excessive Infancy Weight Gain
Authors: Metallinos‐Katsaras Elizabeth Sophia , Siu Elizabeth Colavito , Colchamiro Rachel
Published: June 2021

Objective

To assess the associations between household (HH) food security status (FSS), and excessive (EWG) and inadequate (IWG) weight gain among low income infants.

Sample

infants on WIC (2001–2009) with complete data (n=31,423) on weight and length in early (2 months) and late (12 months) infancy, HH FSS in early (0–6 mos) and late (12 + 2 months) infancy, breastfeeding and covariates.

Outcomes

The following were based on changes in weight‐for‐length z‐scores (WHO) between early and late infancy: 1) EWG = >; 2 z‐score increase 2) IWG= >; −1 z‐score decline.

Analysis

Multivariate Logistic Regression.

Results

EWG: among first born infants, effects of late infancy HH food insecurity (FI) were modified by: 1) early HH FSS (p < .05), 2) whether the infant was breastfed (p < .01). Among subsequently born infants: 1) persistently low HH FSS (vs. persistent food security) increased risk of EWG (AOR=1.29, 95% CI= 1.10–1.52), 2). Among never breastfed infants, late infancy low HH FSS (vs. late food security) increased risk of EWG (AOR=1.24, 95% CI=1.05–1.47). IWG: Among first born infants, a significant interaction between late FSS and breastfeeding (p=.02) indicated that among never (but not ever) breastfed infants, any HH FI in late infancy (vs. food security), significantly elevated risk of IWG (AOR=1.60, 95% CI=1.19–2.16). Breastfeeding may buffer against adverse effects of FI on infant weight gain. Funding: ERS/USDA
Porcine prepubertal obesity is associated with altered adipokine mRNA in visceral and subcutaneous fat
Authors: Zhang Wei , Scheffler Jason , Escobar Jeffery , Reeves Heather , Van Eyk Greg , Fisher Kimberly , Seelenbinder Kirsten , Price Kathryn , Scheffler Tracy , Zhao Xiaoling , Gilbert Elizabeth
Published: June 2021

Our objective was to determine the effect of a high‐fat and high‐carbohydrate diet on body composition and adipokine mRNA in prepubertal pigs. Female 28‐d‐old Premium Genetics 1020 pigs were divided into 4 groups including 1) Control or highfat (HF) diets containing 2) complex carbohydrates (CC) or refined sugar in the form of 3) glucose or 4) fructose. At 56 d, subcutaneous, mesenteric and perirenal fat were collected. Weight gain was similar (P=0.8) among treatments whereas body fat composition increased (P<0.001) from 18% in controls to 28% in HF pigs. Diet (glucose >; control; P=0.03) and fat (perirenal >; subcutaneous; P=0.04) affected adiponectin mRNA. For monocyte chemoattractant protein 1 and tumor necrosis factor α (TNFα), mRNA was greater (P<0.04) in pigs that consumed CC compared with the control diet. Hypoxia inducible factor 1 was greater in perirenal fat as compared with subcutaneous (P<0.001) and mesenteric (P=0.03). The TNFα mRNA was more abundant (P=0.01) in perirenal as compared with mesenteric. Visfatin mRNA was greater in perirenal and mesenteric fat as compared with subcutaneous (P<0.01). Thus, HF diets are associated with increased adipokine mRNA with site‐specific differences.

Grant Funding Source: Virginia Tech Start‐Up Fund
Bromelain prevents mouse testis from tert‐butyl hydroperoxide‐induced dysfunction by inhibiting apoptosis, inflammation, and cellular senscence
Authors: Yeh Yueh‐Chiao , Tsai Ching‐Han , Liu Tsun‐Jui , Lai Hui‐Chin , Wang Li‐Chuan
Published: June 2021

Researches indicated the bromelain, an extract from pineapple stem, has anti‐inflammatory effects in several system models. In contrast, the lipid prooxidant tert‐butyl hydroperoxide (t‐BHP) has been referred to cause oxidative stress‐mediated testicular damage. This study was to investigate the effects of bromelain against t‐BHP‐induced testicular dysfunction. Male mice at 4‐week of age were given vehicle, t‐BHP alone, bromelain alone, or bromelain followed by t‐BHP. After 2 weeks, mice treated with t‐BHP alone displayed reduced sperm counts, elevated apoptotic indexed (upregulation of Bax, downregulation of Bcl‐2, release of cytochrome c to cytosol, activation of caspase‐3, and increase of cleaved caspase‐3 abudance and TUENL positive cells), intensified differentiated expressions of cell‐cycle regulators (p53, p21, p16, and pRb), activated cellular senescence (upregulated SA‐beta‐gal positive‐cells and downregulated telomerase activity), and impacted male fertility (inhibition of testicular FSH hormone receptor and elevation of serum FSH level). Interestingly, bromelain pretreatment could antagonize nearly all of these abnormalities caused by t‐BHP. In conclusion, bromelain could suppress the apoptotic actions, cellular senescence, and infertility in testis. Bromelain may be a potential natural product for ameliorating testicular failure in clinical practice.
Differential antibody responses to HPV16 in cervical and oropharyngeal cancer
Authors: Unger Elizabeth R. , Panicker Gitika , Wong Jessica , Wallstrom Garrick , Haddad Robert , D'Souza Gypsyamber , Posner Marshall , Anderson Karen S.
Published: June 2021

Background

HPV type 16 is associated with the majority of cervical precancers, invasive cervical cancer (ICC), and oropharyngeal cancers (OPC). Pap tests have lower sensitivity and higher specificity than HPV DNA testing for the selective detection of high risk cervical disease, and there are no biomarkers for OPC. Antibodies (Abs) to HPV16‐derived early proteins are potential biomarkers of HPV‐associated malignancies.

Methods

Using a custom bead array ELISA, Abs to the HPV16 proteome were measured in sera from patients with no cervical disease (CIN 0/1; n=121), high‐grade cervical precancer (CIN II/III; n=162), ICC (n=95), and OPC (n=50). The median fluorescent intensity (MFI) ratios of IgG specific for each HPV antigen to control antigen were determined and compared to cut‐off values from 50 healthy donors. HPV DNA detection and typing used L1 consensus PCR with type‐specific hybridization.

Results

At least one HPV16 Ab to E1, E2, E6, or E7 was detected in the sera of 26% of HPV16+ ICC cases, compared with 0% of HPV+ CIN 0/1 controls and 3% of HPV+ CIN II/III and 92% of OPC. In comparison to HPV16+ ICC sera, OPC sera had higher levels of Abs to at least one antigen (p<0.0001).

Conclusion

Serum IgG Abs to multiple HPV16 proteins are detected in HPV associated invasive carcinomas. In contrast to ICC, OPC sera contain strong E1, E2, and E7 Abs, suggesting differences in biology between these two HPV‐associated malignancies.
Mouse models of cancer – how to optimize their predictive value in cancer drug development
Authors: Herter‐Sprie Grit Sophie , Wong Kwok‐Kin
Published: June 2021

Continuously rising incidences of malignancy convert cancer into one of the major threats humankind faces. The current conventional treatment modalities like surgery, chemo‐ and radiation therapy result in encouraging response rates, but antineoplastic agents providing long‐term survival are still missing. Hence, efficient and accelerated translation of potential cancer drugs is urgently needed to antagonize increasing cancer mortality. Indispensable tools in anticancer component development are autochthonous mouse models of cancer and patient‐derived tumor xenografts. However, despite their continuous popularity the probability of successful translation from these preclinical animal models to clinical trials in humans remains low. Here we report recent advances in improved prediction of clinical trial outcome employing “mouse avatars” and co‐clinical trials.
Molecular histology with high resolution imaging mass spectrometry
Authors: Kellersberger Katherine , Kowalski Jane‐Marie , Becker Michael , Deininger Soeren‐Oliver , Cornett Shannon
Published: June 2021

Imaging mass spectrometry (IMS) compliments traditional histopathology by illuminating the biochemical processes that accompany visual changes in cellular structure. As an imaging technique, IMS is characterized by spatial resolution, but as a mass spectrometry technique IMS is characterized by the range of compounds detected. In an effort to improve the performance of IMS for high‐resolution, label‐free molecular histology we investigated two key aspects of IMS: instrumentation (laser focus) and sample processing (analyte diffusion).

Using a solvent‐free preparation, images of phospholipids acquired from rat brain exhibit virtually no analyte diffusion and allow the laser focus to be reduced to cellular dimensions. For imaging larger proteins, however, solvent‐free conditions are not effective and traditional solvent‐based preparations induce substantial analyte diffusion thereby limiting resolution. Rat brain sections were processed in a high‐humidity atmosphere instead of using liquid solvent and a large number of high resolution protein images were collected. This shows that such conditions provide efficient analyte extraction with minimal migration. High resolution IMS images viewed in conjunction with histopathology show a more detailed picture of cellular processes and promises a better understanding of mechanisms of disease progression and treatment response.
Liver microRNAs are differentially expressed in human simple steatosis and non alcoholic steatohepatitis with potential repercussions on lipid metabolism and inflammatory status
Authors: Comelli Elena Maria , Kim TaeHyung , Arendt Bianca M , Singh Natasha , Zhang Zhaolei , Allard Johane P
Published: June 2021

Non‐alcoholic fatty liver disease is associated with fat deposition in liver. It can present as simple hepatic steatosis (SS) or as non alcoholic steatohepatitis (NASH), which is characterized by inflammation leading to fibrosis. It is unclear which molecular mechanisms differentiate SS and NASH. Progression of SS to NASH is associated with altered hepatic microRNA (miRNA) expression in animal models, but human data are scarce. We analyzed the miRNA expression signature in histology‐proven healthy (H; n= 24), SS (n= 24) or NASH (n= 23) human adult livers by NanoString technology. We found that samples from the 3 groups cluster by disease state based on 142 differentially expressed miRNAs, suggesting that a specific liver miRNA signature distinguishes H, SS and NASH. Putative gene targets of these miRNAs were identified with 12 algorithms and used for gene ontology enrichment analysis. This showed that processes including gene transcription, apoptosis and the TGF‐β signaling pathway are commonly altered between H, SS and NASH. In addition, NASH was associated with altered regulation of lipid storage, IL‐6 and adipocytokine signaling pathways as compared to H and SS and regulation of cholesterol storage as compared to SS. These differences may have therapeutic potential.

Funding sources: American College of Gastroenterology, Canadian Institutes of Health Research
Functional Variations of Organic Cation Transporters associated to Inflammatory Bowel Disease (IBD) in a Manitoba Cohort
Authors: Eck Peter Karl , Serrano Alejandra , Yurkova Natasha , Berstein Charles
Published: June 2021

Polymorphisms in SLC22A4, SLC22A23 and IBD5 locus have been associated with pathogenesis of IBD. This study sought to investigate the association of common and rare single nucleotide polymorphisms (SNPs) within those genes in Manitoba's IBD population. DNA samples from 160 patients with Crohn's Disease (CD), 149 patients with Ulcerative Colitis (UC) and 142 healthy controls were genotyped by PCR‐RFLP analysis. The IBD5 rs11739136‐C and rs17622208‐A alleles are associated with increased susceptibility to CD (OR=1.8, 95%CI 1.28–2.5, p=0.0006; OR=1.4, 95%CI 1.01–1.92, p=0.04, respectively). Moreover, rs11739136‐CC homozygosity increased the susceptibility of UC (OR=4.74, 95% CI 1.74–12.9, p=0.0009). Both CD (62%) and UC (64%) showed higher frequencies of SLC22A23 rs17309827 T allele than controls (61%).

SLC22A4 rs1050152 showed strong Linkage Disequilibrium (LD) (D′ =0.98) with rs11739135. The risk alleles conform a haplotype (TACT) which is carried by 38.8% in CD patients and 29.8% in controls (p=0.02). We report that rs11739135 and rs17622208 best tag IBD5 haplotypes.

Two selected rare polymorphisms in SLC22A4 (rs11568500 and rs11568510) were not detected in our population. Rare variations might be relevant in populations with different ethnical background. Further studies on the role of solute transporters in IBD are needed to develop therapeutic interventions.
Endothelial dysfunction accentuates the loss of function of beta adrenergic receptors in the smooth muscle of the rat aorta during maturation
Authors: Lopez‐Canales Oscar , Lopez‐Canales Jorge , Guevara‐Balcazar Gustavo , Castillo‐Hernandez Maria Carmen , Escalante‐Acosta Bruno
Published: June 2021

It is well established that as age increases rat, the relaxing function of beta‐adrenergic receptors is lost gradually, however, the specific involvement of smooth muscle during this process is not yet well established. In an attempt to provide additional information on the topic, we evaluate the beta adrenergic relaxation response in an in vitro model, with two groups: rats of 3 and 9 weeks (maturation period) using arteries with and without endothelium. Our results indicate that during this period of maturation the relaxation obtained with isoproterenol decreases slightly (90 ± 3% vs 81 ± 2%) in aortic rings with endothelium, whereas the response in rings without endothelium the decrease is much larger (87 ± 4% vs 21 ± 2%). The next step was to try to establish that molecular changes smooth muscle suffers during this period, to this, there were concentration‐response curves in the presence of a wide range of inhibitors, and the results indicate that the signaling pathway used by these receptors does not change with age. In conclusion, the absence of endothelium increases the loss of function of beta adrenergic receptors, this effect cannot be explained by a change in the signaling pathway, which probably explained by changes in the expression and translation rate of the molecules involved.
Additive antiemetic efficacy of Δ9‐THC with vanilloid TRPV1 receptor agonists in the least shrew (Cryptotis parva)
Authors: Brar Rajivinder Singh , Chebolu Seetha , Zhong Weixia , Darmani Nissar A.
Published: June 2021

Previous studies have shown that cannabinoid CB1 and vanilloid TRPV1 agonists ( Δ9‐THC and resiniferatoxin (RTX), respectively) can attenuate the emetic effects of chemotherapeutic agents such as cisplatin. In this study we used the least shrew to demonstrate whether combination of varying doses of Δ9‐THC with resiniferatoxin can produce additive antiemetic efficacy against cisplatin‐induced vomiting. RTX by itself caused vomiting in a bell‐shaped dose‐dependent manner with maximal vomiting at 18 ug/kg when administered sc but not ip. Δ9‐THC up to 10 mg/kg can only protect least shrews from cisplatin‐induced emesis by 80% with an ID50 of 0.3 – 1.8 mg/kg. Combinations of 1 or 5 ug/kg RTX with varying doses of Δ9‐THC completely suppressed both the frequency and the percentage of shrews vomiting with ID50 dose values 5–50 times lower than Δ9‐THC doses tested alone against cisplatin. A less potent TRPV1 agonist capsaicin, by itself did not cause emesis (ip or sc), but did significantly reduce vomiting induced by cisplatin given after 30 minutes but not at 2 hours. TRPV1 antagonist ruthenium red attenuated cisplatin‐induced emesis at 5 mg/kg, however capsazepine did not. In summary, we present evidence that combinations of CB1 and TRPV1 agonists have the capacity to completely abolish cisplatin‐induced emesis at doses lower than when used individually.
Interactions between mu opioid receptor agonists and cannabinoid receptor agonists CP55940 and WIN55212–2 in rhesus monkeys: evaluation of treatment‐ and abuse‐related effects
Authors: Maguire David R , Yang Wenjuan , France Charles P
Published: June 2021

Mu opioid receptor agonists and cannabinoid receptor agonists can be combined to possibly enhance therapeutic effectiveness (e.g., pain treatment); however, it is unclear whether combinations of these drugs enhance abuse liability. These studies examined the effects of cannabinoid receptor agonists CP55940 and WIN55212‐2 on the antinociceptive, discriminative stimulus, and positive reinforcing effects of mu opioid receptor agonists in rhesus monkeys. In one group (n=3), CP55940 (0.001–0.056 mg/kg), and WIN55212‐2 (0.1–1.78 mg/kg) shifted morphine dose‐effect curves leftward up to 8 fold. In monkeys discriminating 3.2 mg/kg morphine from saline (n=3), CP55940 and WIN55212‐2 shifted the morphine discriminative stimulus dose‐effect curves rightward up to 7 fold. In monkeys (n=4) self‐administering heroin (0.32–32.0 μg/kg/infusion), CP55940 and WIN55212‐2 shifted the heroin dose‐effect curves rightward (up to 15 fold) and downward. Cannabinoid receptor agonists CP55940 and WIN55212‐2 enhanced the antinociceptive effects but not the discriminative stimulus or positive reinforcing effects of mu opioid receptor agonists. These data support the view that combining mu opioid receptor agonists and cannabinoid receptor agonists might enhance treatment effectiveness without enhancing abuse liability. Supported by USPHS Grants R01DA05018, K05DA017918 (CPF) and T32DA031115 (DRM).
Induction of CYP2D6 Expression during Pregnancy Is Associated with an Increased Activity of Hepatocyte Nuclear Factor 4α
Authors: PAN XIAN , Koh Kwihye , Yu Ai‐Ming , Jeong Hyunyoung
Published: June 2021

Clinical evidence indicates increased metabolism of CYP2D6 substrates during pregnancy; the underlying mechanisms are unknown. The objectives of this study were (1) to evaluate the utility of CYP2D6‐humanized transgenic (Tg‐CYP2D6) mouse as a model for CYP2D6 induction during pregnancy, and (2) to investigate the role of hepatocyte nuclear factor 4α (HNF4α), a known transcriptional activator of CYP2D6, in CYP2D6 induction. The genome of Tg‐CYP2D6 mouse harbors human CYP2D6 gene with its 2.5‐kb upstream regulatory region. qRT‐PCR and western blot results revealed that CYP2D6 mRNA and protein expressions was 2–3 fold higher in Tg‐CYP2D6 mice during pregnancy than pre‐pregnancy, consistent with an elevated CYP2D6 metabolism shown in pregnant women. In contrast, mRNA expression of Cyp2d22, the mouse homologue of human CYP2D6, did not increase during pregnancy. While hepatic HNF4α mRNA or protein levels were not altered in Tg‐CYP2D6 mice at pregnancy, ChIP‐qPCR study demonstrated an increased recruitment of HNF4α (and RNA polymerase II) onto CYP2D6 promoter region (~3 fold) in livers of pregnant mouse vs. those of virgin or postpartum mice. An increased recruitment was also shown for another HNF4α target gene, ApoC2. Our results suggest that CYP2D6 induction during pregnancy may be due to the increase of HNF4α activity that enhances transcription expression of CYP2D6. This study is supported by NIH (R01HD065532).
Pterostilbene Inhibits VEGF Production in Human Breast Cancer [MDA‐MB‐231] Cells
Authors: Sanghani Arpitkumar , Kandra Theja , Bavadekar Supriya , Vansal Sandeep
Published: June 2021

Pterostilbene, a stilbenoid and natural analog of resveratrol, is present mainly in blueberries, and has been shown to have anticancer, anti‐inflammatory, and antioxidant properties in various cancer cell lines. However the anti‐angiogenic potential of pterostilbene has not yet been established in breast cancer. The objective of this research was to investigate the effect of pterostilbene on pro‐angiogenic factors including vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP‐2 and MMP‐9), and inflammatory cytokines (IL‐6 and IL‐8) in human metastatic, breast cancer cells, MDA‐MB‐231. The cells were treated with increasing concentrations of pterostilbene ranging from 10 μM to 100 μM for 24hr, and secreted VEGF concentrations were measured using ELISA. Pterostilbene caused a dose‐dependent decrease of secreted VEGF [~45% inhibition at 100 μM] when compared with control. Further studies are underway to understand the molecular mechanism involved. These results indicate that pterostilbene possesses anti‐angiogenic properties that may critically contribute to its observed anticancer effects. This work is supported by the Division of Pharmaceutical Sciences at Long Island University and the Touro College of Pharmacy.
The Ameliorative Effect of Silymarin on Cadmium‐Treated Human Lung Carcinoma Cells
Authors: Webster Camille A , Latinwo Lekan M. , Badisa Veera L.D. , Odewumi Caroline O. , Darling‐Reed Selina F.
Published: June 2021

Purpose

Cadmium is a non‐essential metal and environmental hazardous pollutant. The exposure of cadmium causes damage to all organs of human body and mainly accumulates in the liver and lungs due to cigarette smoke that contains cadmium. This leads to cancer in different organs of the body. Silymarin is a polyphenolic antioxidant flavonoid complex found in the herb, Silybum marianum (milk thistle seed).

Methods

In this study, the ameliorative effect of silymarin on the viability and the DNA damage was evaluated in cadmium‐induced human lung carcinoma cells (A549) by comet assay.

Results

The results showed that treatment of human lung carcinoma cells with cadmium chloride alone for 24 h resulted in decrease in cell viability; while co‐treatment with silymarin showed an increase in viability. The human lung carcinoma cells treated with cadmium chloride alone increased the tail DNA and tail length; while co‐treatment with silymarin decreased the tail DNA and tail length in both single and double strand DNA comet assays.

Conclusion

Our results clearly show the ameliorative effect of silymarin on the viability and DNA damage in human lung carcinoma cells and suggest that it can be used for prevention and treatment of cadmium toxicity.
Development of selective inhibitors of the isoprenoid biosynthetic pathway and their effects on human‐derived chronic myelogenous leukemia cells
Authors: Reilly Jacqueline E , Zhou Xiang , Tong Huaxiang , Wiemer David , Hohl Raymond J
Published: June 2021

Proteins dependent on post‐translational geranylgeranylation by the isoprenoid biosynthetic pathway (IBP) are key regulators of malignant cell properties, such as migration, invasion, and proliferation. Nonspecific IBP inhibitors, including statins, have beneficial effects on these malignant cell properties. However, they broadly deplete all mevalonate derivatives and thus affect farnesylated proteins and cholesterol. We have developed two isoprenoid bisphosphonates, XZ‐1–269 and XZ‐1–277, to selectively inhibit geranylgeranyl diphosphate synthase (GGDPS) and diminish only geranylgeranylated proteins. In K562 human‐derived chronic myelogenous leukemia cells XZ‐1–269 impairs geranylgeranylated proteins at 2.5μM as compared to 10μM for digeranyl bisphosphonate (DGBP), and reduces cell viability below 80% at 0.1μM as compared to 10μM for DGBP at 72hrs. Comparatively XZ‐1–277 is inactive, explained by the examination of the crystalline structure of GGDPS. Our findings present a novel, potent compound for selectively reducing geranylgeranylated proteins. Expansion of this family with in silico and experimental structure‐activity relationships will allow for development of even more potent IBP inhibitors. NIH Pharmacological Sciences Training Grant T32 GM067795.
Fibroblast growth factor 21 is required for alcohol‐induced adipose tissue lipoatrophy in mice
Authors: Liu Yanlong , Zhao Cuiqing , Wang Yuhua , McClain Craig , Feng Wenke
Published: June 2021

Fibroblast growth factor 21 (FGF‐21) is a member of FGF superfamily and has been identified as a potent metabolic regulator. Studies have shown that FGF‐21 plays a critical role in adipose tissue lipolysis. FGF‐21 KO mice and controls (C57BL/6 mice) were fed Lieber deCarli liquid diet containing 5% alcohol or pair‐fed with isocaloric maltose dextrin for 12 days. Six hours before sacrificing, a binge dose of alcohol at 6g/kg body weight was given to the mice by gavage. Alcohol significantly increased serum and hepatic FGF‐21 in wild type mice. Alcohol decreased white adipose tissue mass in wild type mice, but not in FGF‐21 KO mice, indicating a requirement of FGF‐21 for alcohol‐induced adipose tissue lipoatrophy. A mechanism underlying adipose tissue lipolysis is the cAMP‐mediated hormone sensitive lipase (HSL) pathway, which is regulated by catecholamines, nicotinic acid, and insulin. Perilipin (PLIN) is a coating protein for lipid droplet and protects lipids waiting to be degraded. Alcohol increased phosphorylation of PLIN and HSL in wild type mice, which may be responsible for the observed adipose lipolysis by alcohol. Alcohol did not affect PLIN and HSL phosphorylation in FGF‐21 knockout mice. Our results suggest that FGF‐21 is a potential regulator of alcohol‐induced white adipose tissue hyperlipolysis via HSL/PLIN pathway, which may contribute to alcoholic fatty liver formation.
Decreased transglutaminase activity in aorta from hypertensive rats, measured by in situ detection of a free amine donor
Authors: Petersen‐Jones Humphrey G , Johnson Kyle B , Watts Stephanie W
Published: June 2021

Transglutaminases (TGs) catalyze protein modifications in vasculature by forming covalent bonds between protein‐bound glutamine residues and free amine groups. TG protein remodeling has been implicated in hypertension. Studies measuring in situ TG activity with glutamine donor peptides K5 (TG1) and T26 (TG2) show a decrease in TG1 and TG2 activity in aorta from hypertensive rats vs normotensive rats. To measure TG activity we used Pentylamine‐Biotin (BaP), a free amine TG substrate. We hypothesized in situ TG activity will be decreased in aorta from hypertensive rats vs normotensive rats. Using aorta from deoxycorticosterone acetate (DOCA)‐salt, sham, spontaneously hypertensive stroke‐prone (SHRSP), and Wistar‐Kyoto (WKY) rats, (mean BP 212, 115, 202, and 117 mmHg, respectively) we performed in situ detection of TG activity using BaP visualized with Alexa Fluor 555‐linked streptavidin. We observed decreased TG activity in aorta from hypertensive rats, versus normotensive (DOCA: 0.47±0.034, sham: 0.92±0.047; SHRSP: 0.76±0.077, WKY: 1.4±0.13 mean fluorescence/mm2; p<0.05; n=4). We are perfecting a protein assay using biotinylated K5 and T26 peptides to identify TG1 and TG2 specific substrates. In conclusion, TG activity is decreased in aorta from hypertensive rats vs normotensive rats, suggesting a decrease in TG activity plays a role in the pathology of hypertension.

The APS supported this research.
A pivotal role for pericytes in non‐steroidal anti‐inflammatory drug‐induced toxicity
Authors: Peppiatt‐Wildman Claire M , Kennedy‐Lydon Teresa , Crawford Carol , Wildman Scott S P
Published: June 2021

Pericytes regulate vasa recta diameter in response to numerous endogenous vasoactive agents [1] and in so doing regulate medullary blood flow (MBF). Renal pericytes have also been implicated in numerous forms of kidney disease. Non‐steroidal anti‐inflammatory drugs (NSAIDs), routinely prescribed to alleviate pain and inflammation, have established nephrotoxic side effects, such as reduced MBF, that are often associated with chronic renal disease. NSAIDs inhibit cyclooxygenase (COX) enzymes and the production of endogenous vasodilatory prostanoids, which we have previously shown to exert their effects on vasa recta diameter via their action at pericytes. We propose renal pericytes are pivotal in NSAID‐mediated reduction in MBF and associated nephrotoxicity.

We have utilized the live kidney slice model [1] to demonstrate NSAID‐mediated reduction in vasa recta diameter occurs specifically at pericytes and have identified NSAID‐mediated inhibition of vasodilatory PGE2 and PGI2 as critical in this process. We identify renal tubules as the source of vasodilatory prostanoids and suggest pericyte‐mediated tubulovascular cross‐talk mechanisms are vital for MBF regulation, and dysregulation of tubulovascular cross‐talk is central in NSAID‐associated nephrotoxicity.
Role of connexin 45 in the autoregulatory response of the afferent arteriole
Authors: Sorensen Charlotte Mehlin , Braunstein Thomas Hartig , Holstein‐Rathlou Niels‐Henrik
Published: June 2021

Connexins expressed in renal arterioles are believed to influence the regulation of arteriolar tonus and renal blood flow. Connexin 45 (Cx45), which is primarily expressed in vascular smooth muscle cells of the renal afferent and efferent arterioles, has been suggested to play a role in the regulation of renin secretion. Also, Cx45 may be involved in the transmission of the signal initiating the tubuloglomerular feedback (TGF) response.

In mice lacking Cx45 expression in vascular smooth muscle cells we examined the renal autoregulatory response by measuring changes in afferent arteriolar diameter in response to step increases in perfusion pressure. We used the isolated blood‐perfused juxtamedullary nephron preparation to visualize afferent arterioles. Cx45 deficient mice did not seem to have a higher mean arterial blood pressure than age‐matched litter‐mate wild type controls. The reduction in afferent arteriole diameter in response to step increases in renal perfusion pressure was significantly reduced in Cx45 deficient mice compared to the wild‐type litter‐mates. Removal of the TGF response did not seem to affect the ability to regulate afferent diameter in Cx45 deficient animals compared to wild‐type litter mates. These results suggest that Cx45 plays a major role in the myogenic component of the autoregulatory response of the afferent arteriole.