bluuebunny/crossref_metadata_2025 huggingface dataset:
BMI Distribution and Body Image of Korean High School Students in Gyeonggi Province
Authors: Chang Kyung Ja , Lee Suk‐young
Published: June 2021

In order to investigate BMI distribution and body image of Korean high school students for nutrition education, a cross‐sectional survey was conducted using a questionnaire. The subjects were 493 high school students (male 241, female 252) in Ansan, Gyeonggi Province, Korea. The data were analyzed with SPSS 12.0 program. According to the standard of the Korean Obesity Association, the students whose BMI was below 18.5 as underweight, between 18.5 and 22.9 as normal, between 23 and 24.9 as overweight, and 25 or more as obese. As for the distribution of weight by BMI, there was a significant difference between male and female students; 41.1% of the male normal, 31.5% overweight, 14.5% underweight, 12.9% obese and 51.6% of the female normal, 26.5% overweight, 16.3% underweight, 5.6% obese(p<0.01). As for body image, there was a significant difference between male and female students; 44.2% of the male students who have normal weight perceive themselves as fat, whereas 61.6% of the female students who have normal weight perceive themselves as fat(p<0.001). As for an ideal body shape, there was a significant difference between male and female students; 80% of the male students answered to have normal figure, 13.7% did thin figure, while 59.1% of the female students answered to have normal figure, 37.7% did thin figure(p<0.001). Therefore, personalized nutrition education by gender should be provided to correct distorted body image of high school students in Gyeonggi Province, Korea.
Common fSNP variants of fourteen Bardet‐Biedl syndrome genes and adult body mass
Authors: Birk Ruth Z , Ermakov Sergey , Livshits Gregory
Published: June 2021

Bardet‐Biedl syndrome (BBS) is a rare monogenic multi‐systemic disorder manifesting with marked obesity. Fourteen BBS genes have been identified to date and additional loci are expected. Mutations of several BBS genes were shown to affect fat cell differentiation. To date, association between common polymorphisms in all 14 genes as a group and body weight has not been tested.

Study objective and method

We investigated association between tagging single nucleotide polymorphisms (tSNPs) of 14 BBS genes, and body weight and fat in 2462 adult women from the UK Twins study. Significant results were further tested in a confirmation sample of 2003 women from the same cohort and additionally in the GIANT consortium population (n=123,865).

Results

105 SNPs in 14 BBS genes were selected and tested in the first cohort for association with body weight and fat phenotypes, i.e. weight, BMI, total body fat (assessed by DEXA), total fat/height2, and total fat/weight. We used principal component (PC) derived using the latter five traits as a primary phenotype for this study. Of the 105 SNPs, 3 variants in BBS9 and BBS11 showed evidence of nominally significant association with elevated body weight and fat. However, none of the associations survived multiple‐testing correction.

Conclusion

The results suggest that common variation in 14 BBS genes are unlikely to have a substantial effect on body weight and fat in the European population
Effect of hypercaloric diet on RAGE mRNA expression, oxidative stress and kidney function in rats
Authors: Pierine Damiana Tortolero , Luvizoto Renata Azevedo Melo , Francisqueti Fabiane Valentini , Ferreira Ana Lucia Anjos , Yeum Kyung‐Jin , Correa Camila Renata
Published: June 2021

Advanced glycation end products (AGEs) produced under oxidative stress can trigger porinflammatory responses by interacting with receptors for AGEs (RAGE). RAGE may link to the pathogenesis of chronic kidney diseases.

Objective

To evaluate the influence of hypercaloric diet on RAGE mRNA expression and oxidative stress in the kidney and its physiologic function.

Methods

Male Wistar rats were fed a control diet (C, n=14) or a hypercaloric diet (HD, n=14) for 12 weeks. The adiposity index (AI) was used as an obesity indicator. Kidney function was evaluated by glumerular filtration rate, kidney RAGE mRNA expression by real time PCR, and kidney oxidative stress by total antioxidant performance (TAP).

Results

RAGE mRNA expression was significantly (p<0,001) higher in HD (1.89±0.77) than C rats (0.76±0.37). A positive correlation between RAGE and AI (r=0.64, p <0.001) was found. There were no differences in glomerular filtration rate and TAP between groups.

Conclusion

The hypercaloric diet for 12 weeks resulted in an increase of RAGE mRNA expression, which may lead to inflammation in the kidneys from HD group.

Support: CAPES, FAPESP (2010/06100–9)
Precursors for ornithine and citrulline synthesis in neonatal pigs
Authors: Marini Juan C , Stoll Barbara , Didelija Inka Cajo , Burrin Douglas G
Published: June 2021

Citrulline (CIT) is an amino acid synthesized by gut and utilized for the synthesis of the conditionally essential amino acid arginine (ARG). In turn, the immediate precursor for CIT synthesis, ornithine (ORN), can originate from proline (PRO) and glutamine (GLN) via ornithine aminotransferase (OAT, i.e., de novo synthesis) or from ARG via arginase. To test the hypothesis that during the neonatal period de novo synthesis is the main source of ORN for CIT synthesis, neonatal pigs were infused iv or ig with [U‐13C6]ARG, [U‐13C5]GLN or [U‐13C5]PRO during the fasted and fed periods. (ureido)[15N]CIT and [D2]ORN were infused iv for the entire infusion protocol. During fasting plasma PRO (13%) and ORN (19%) were the main precursors for CIT synthesis, whereas plasma ARG (62%) was the main precursor for plasma ORN. During feeding enteral (27%) and plasma (12%) PRO were the main precursors for the ORN utilized in the synthesis of CIT, together with plasma ORN (27%). Enteral PRO and GLN were utilized directly by the gut to produce ORN utilized for CIT synthesis. ARG was not utilized by the gut, which is consistent with the lack of arginase activity in neonates. ARG, however, was the main source (47%) of plasma ORN and in this way contributed to CIT synthesis. During the neonatal period the directionality of OAT, together with the lack of arginase activity in the gut, determine that the de novo pathway is the main source of ORN for CIT production.
Effects of iron deficiency on the expression of insulin‐like growth factor II and its receptor (IGF‐IIR) in murine glial cells
Authors: Morales‐González Esmeralda , Contreras Irazú , Estrada José Antonio
Published: June 2021

Objective

To determine the expression of IGF‐II and IGF‐IIR on iron‐deficient murine glial cells.

Methods

Primary glial cultures were established from purified cells obtained from the brain of newborn BALB/c mice. Mixed glial cultures were plated at a density of 120,000 cells/cm2 in complete DMEM (10% FBS) and cultured for 14 days. Subsequently, half of the cells were treated with deferoxamine (100 μM/mL) for 24 hours, to simulate iron‐deficient conditions. After treatment, cells were lysed and their protein contents extracted. Protein concentrations were assessed by the Bradford method and adjusted at 1 μg/μL. 70 μg of protein per sample were used to detect expression of IGF‐II and IGF‐IIR by western‐blot.

Results

We detected decreased expression of IGF‐IIR in cells treated with deferoxamine, compared with controls. No difference was observed in the expression of IGF‐II under the same conditions.

Conclusion

Iron deficiency decreases the expression of IGF‐IIR, but not IGF‐II, in murine mixed glial cultures. Decreased expression of this receptor may have important consequences for the development and function of the central nervous system during iron deficiency.
Pharmacokinetics of a novel eicosapentaenoic acid derived anti‐leukemic prostaglandin, Δ12‐ prostaglandin J3 in a murine model
Authors: Kudva Avinash Kundadka , Kaushal Naveen , Paulson Robert F , Prabhu Sandeep K
Published: June 2021

Cyclopentenone prostaglandins are downstream metabolites of prostaglandin D series that exhibit anti‐inflammatory and anti‐carcinogenic activities. Recently, our laboratory has shown that Δ12‐prostaglandin J3 (Δ12‐PGJ3), produced from dietary omega‐3 polyunsaturated fatty acid (n‐3 PUFA), eicosapentaenoic acid (EPA), alleviates the progression of leukemia in three murine models with a remarkable selectivity towards leukemia stem cells (LSC), but not the normal hematopoietic stem cells (HSC). In the present study, we have evaluated the stability and pharmacokinetic properties of Δ12‐PGJ3in vivo models to establish efficacy and toxicology profile enabling better therapeutic strategies in the future. These studies should have a bearing on the nutritional intervention strategies using EPA‐rich diets in the above models of the disease.

Grant Funding Source: NIH
Influence of Diabetes on Plasma Pharmacokinetics and Brain Bioavailability of Grape Polyphenols in the Zucker Rat Model
Authors: Chen Tzu‐Ying , Janle Elsa M , Ferruzzi Mario G , Simon James E , Wu Qing‐Li , Talcott Stephen T , Weaver Connie , Wang Jun , Ho Lap , Pasinetti Giulio M
Published: June 2021

Grape polyphenols (GP) have been demonstrated to prevent Alzheimer's disease (AD) in a mouse model. Patients with diabetes mellitus are at greater risk of developing AD than non‐diabetic patients. To investigate the effect of the diabetic condition on bioavailability and brain distribution of flavan‐3‐ols, flavonols, and anthocyanins, Zucker diabetic fatty (ZDF) rats and their lean control (LN) were dosed orally with a combination of GP from grape seed extract, Concord grape juice and resveratrol (RES). Eight hour plasma pharmacokinetics was conducted after oral gavage of GP or water for 10 days. LC‐MS/MS analysis identified 3‐glucuronide (3‐glucur) and 3′‐methylglucuronide (3′‐OMe‐glucur) as the main metabolites for catechin (C), epicatechin (EC), quercetin (Q), and RES present in plasma and brains. ZDF rats exhibited significantly diminished maximum plasma concentrations for all above metabolites. ZDF rats showed significantly lower bioavailability by 77%, 79% and 45% in 3′‐OMeC‐glucur, 3′‐OMeEC‐glucur and RES‐3‐glucur relative to the LN. ZDF rats exhibited significantly lower levels than LN in all above metabolites except for Q‐3‐glucur in brain tissues. This study showed that the diabetic condition may impact absorption and distribution of polyphenols, suggesting that dosages required to reach therapeutic efficacy may need to be adjusted for disease condition.

Grant Funding Source: NIH Grant P01AT004511
Vitamin C status is inversely related to quercetin bioavailability in young adults
Authors: Guo Yi , Mah Eunice , Jalili Thunder , Bruno Richard S
Published: June 2021

Defining factors regulating quercetin (Q) bioavailability may help to understand its cardioprotective activities. We conducted a pharmacokinetic trial to investigate whether vitamin C status affects Q bioavailability in young adults. Participants (n = 7M/5F, 21.3 ± 3.2 y, 22.8 ± 4.9 kg/m2) ingested 1095 mg Q aglycone with a standardized meal. Plasma vitamin C and endotoxin were measured prior to ingesting Q whereas Q and its methylated metabolites, isorhamnetin (ISO) and tamarixetin (TAM), were measured at timed intervals for 24 h. Maximal plasma Q (1.21 ± 0.66 μM) occurred at 6.0 ± 3.3 h, Q half‐life was 12.2 ± 7.2 h, and inter‐subject variability of Q AUC0–24 h (15.51 ± 6.44 μM * h) was 42%. Plasma Q AUC0–24 h was related to AUC0–24 h of ISO and TAM (r = 0.91–0.96), indicating that greater Q bioavailability increases its biotransformation. Plasma vitamin C was inversely related to the AUC0–24 h of Q, ISO, and TAM (r = −0.58 to −0.72), suggesting that poor vitamin C status increases Q bioavailability and its propensity to be methylated. Vitamin C was also inversely related to endotoxin (r = −0.58), and endotoxin was correlated with total Q (Q + ISO + TAM; r = 0.50), suggesting that poor vitamin C status increases Q bioavailability by increasing intestinal permeability. Future dietary interventions are warranted to investigate whether alterations in vitamin C status regulate Q bioavailability. Funded by Donaghue Nutrition Research Program.
Beta cell dysfunction during hyperglycemia: protective role of erythritol?
Authors: Boesten Danielle , den Hartog Gertjan , de Cock Peter , Bosscher Douwina , Bast Aalt
Published: June 2021

Diabetes is characterized by an alteration of insulin secretory function. The main cell type affected is the beta cell, the only insulin producing cell in the body. Hyperglycemia, as occurring during diabetes, has a harmful effect on the function and development of beta cells. Consequently, chronic hyperglycemia will result in beta cell dysfunction and ultimately cell death. In previous studies erythritol, a polyol bulk sweetener, has been shown to prevent loss of ex vivo endothelium dependent vasorelaxation in streptozotocin diabetic rats and protect endothelial cells during exposure to high concentrations of glucose in vitro1. Extending these studies, we investigated the potential of erythritol to protect beta cells against glucose induced cell death in two cell lines, the hamster beta cell line HIT‐T15 and the human beta cell line 1.1E7. Additionally, we investigated the insulin secretory function of the beta cells and determined the expression of genes involved in apoptosis and insulin secretion.

Research financially supported by Cargill Inc.
Quercetin induces growth arrest through transcription factor FOXO1 in EGFR‐overexpressing oral cancer
Authors: Huang Chun‐Yin , Chan Chien‐Yi , Chou I‐Tai , Lien Chia‐Hsien , Lee Ming‐Fen
Published: June 2021

The squamous cell carcinomas of the head and neck (SCCHN) with aberrant epidermal growth factor receptor (EGFR) signaling are often associated with poor prognosis and low survival. Therefore, efficient inhibition of the EGFR signaling could intervene with the development of malignancy. Quercetin appears anti‐tumorigenesis, but the underlying mechanism is unclear in oral cancer. Fork‐head box O (FOXO) transcription factors, Akt downstream effectors, are important regulators of cell growth. Here, we hypothesized that FOXO1 might be crucial in quercetin‐induced growth inhibition in EGFR‐overexpressing oral cancer. Quercetin treatment suppressed cell growth by inducing G2/M arrest and apoptosis in EGFR‐overexpressing HSC‐3 and TW206 oral cancer cells. Quercetin inhibited EGFR/Akt activation with a concomitant induction of FOXO1 activation. FOXO1‐knockdown attenuated quercetin‐induced p21 and FasL expression and subsequent G2 arrest and apoptosis, respectively. Likewise, quercetin suppressed tumor growth in HSC‐3 xenograft mice. Taken together, our data indicate that quercetin is an effective anti‐cancer agent, and that FOXO1 is crucial in quercetin‐induced growth suppression in EGFR‐overexpressing oral cancer.

This work was supported by the grants from China Medical University CMU97–303 and National Science Council NSC98–2320‐B‐039–021‐MY3
Increases in plasma lutein and zeaxanthin by egg intake is associated with the formation of larger HDL particles in participants with metabolic syndrome
Authors: Blesso Christopher N , Andersen Catherine J , Bolling Bradley , Fernandez Maria Luz
Published: June 2021

We investigated the effects of egg intake on lipoprotein subfractions and size and on plasma lutein and zeaxanthin in metabolic syndrome. Men and women following a carbohydrate restricted diet (25–30% energy) were allocated to consume 3 eggs per day (additional 534 mg of cholesterol, 427 ìg of lutein, 265 ìg of zeaxanthin) (n = 20) or the equivalent amount of egg substitute (additional 0 mg of cholesterol and carotenoids) (n =17) for 12 wk. All subjects increased large LDL (P < 0.05) and decreased small LDL subfractions (P < 0.01). However, increases in the large HDL subfraction were only observed in individuals in the egg group (P < 0.05) Post‐intervention, the egg group had increases in lutein (+ 21%) and zexanthin (+ 43%) while the substitute group had a slight decrease in these plasma carotenoids. Increases in plasma lutein and zeaxanthin were disproportionately due to increases in these carotenoids transported by HDL. We conclude that the higher concentrations of these polar carotenoids in HDL in the egg group may be attributed to a more efficient transport of lutein and zeaxanthin in larger HDL particles [Supported by the American Egg Board]
Fortification of sorghum sorghum vulgare with carrot Daucus carota in the production of sorghum flour
Authors: Banjoko Immaculata Oyeyemi , Oyelola Taiwo Oluwakemi , Onanuga Titilope Tawakalitu , Kujore Fowotade Mutiat
Published: June 2021

The use of cereal preparations in different forms as weaning foods is very common in Nigeria, West Africa. However, some cereals are deficient in vitamin A which has become a global concern in developing countries. Sorghum flour was produced and fortified with carrot powder in an attempt to satisfy this need. Different blends of this preparation were produced in the following ratios of sorghum and carrot respectively. Sample A (100:0w/w); B (90:10w/w); C (80:20w/w) and D (70:30w/w). Physiochemical tests and functional properties were determined by standard methods of analysis while β‐carotens was determined by the use of high performance liquid chromatography (HPLC) technique. Results indicate that all functional and physiochemical parameters were all within the Food and Agricultural Organization (FAO) standard for food fortification. Sensory evaluation using 9‐point hedonic scale showed that sample D(70:30w/w) with a β‐carotene value of 66.1μg was more preferred in terms of taste and overall acceptability.
Community mobilization activities for the Zambia 2012 Orange Maize Efficacy Trial
Authors: Kaliwile Chisela , Arscott Sara A. , Masi Cassim , Tanumihardjo Sherry A.
Published: June 2021

In some societies, blood drawing, vaccinations, and supplementation, are surrounded by myths and beliefs. If not clarified, they may affect study implementation and negatively impact well‐intended studies. Against this background, the implementation team of the “2012 Zambian Orange Maize Study” undertook community mobilization activities to dispel myths, including meetings, drama performances, critical collaborations with local government, and continuous monitoring. The primary aim, assessment of vitamin A status of Zambian children in response to feeding orange maize, was communicated to all stakeholders. This study followed a similar study conducted in the same communities in 2010 and the differences and current study design were discussed. Other activities included renovation of intervention sites, sharing of previous results, staff recruitment, equipment inventory, and phase‐out strategy development. Signing a memorandum of understanding between the stakeholders was critical. Blood drawing, remuneration, and menu items were a source of complaints by parents and slight adjustments were made to the menu based on parental feedback. In conclusion, with all the challenges encountered, the study was successfully implemented according to schedule. Subject participation was good, with few losses to follow‐up. Analysis of the quantitative data is on‐going.

Funded by HarvestPlus.
13′‐Carboxychromanol, a long‐chain metabolite of delta‐tocopherol, has potent anti‐cancer effects by interrupting de novo sphingolipid synthesis in human cancer cells
Authors: Jang Yumi , Kuah Soo Yee , Jiang Qing
Published: June 2021

Natural forms of vitamin E are metabolized to long‐chain carboxychromanols and their sulfated counterparts and the carboxychromanols have been shown to have anti‐inflammatory activities. Although long‐chain carboxychromanols were recently reported to induce apoptosis in hepatocarcinoma HepG2 cells, whether these vitamin E metabolites are effective anticancer agents against other types of cancer remains to be determined. Here we showed that 13′‐carboxychromanol (13′‐COOH), a long‐chain metabolite of delta‐tocopherol (δT), inhibited the growth and induced death of human pancreatic (PANC‐1), breast (MCF‐7) and colon (HCT‐116, HT‐29) cancer cells with IC50 of 6–8 μM, which was much more potent than tocopherols and even gamma‐tocotrienol. Using liquid chromatography tandem mass spectrometry (LC‐MS‐MS), we found that 13′‐COOH induced accumulation of intracellular dihydroceramides and dihydrosphingosin, which are sphingoid bases in the de novo synthesis of sphingolipid pathway. Importantly, this alteration of sphingolipids occurred as early as after 2‐h incubation with 13′‐COOH and therefore preceded its induction of cell death. Meanwhile, an increase of ceramides was observed after 8‐h treatment when cell death became obvious. Our studies demonstrate that 13′‐COOH has more potent anti‐cancer effect than unmetabolized vitamin E forms by modulating sphingolipid metabolism in cancer cells.

Grant Funding Source: R21CA152588 and R01AT006882 from National Institutes of Health
Anti cancer effects of Cnidium officinale Makino extract mediated through Apoptosis and Cell cycle arrest in Human Colorectal cancer cell line (HT‐29)
Authors: dela Cruz Joseph Flores , Kim Gi Dae , Hwang Seong Gu
Published: June 2021

Natural products have recently attracted many researchers due to their potential use as therapeutic agent. Cnidium officinale Makino (COM) has multiple anti‐cancer properties, but its molecular targets and mode of action on human colorectal cancer cells are unknown. Through cell counting kit assay (CCK‐8), dose dependent cytotoxic effects were shown in HT‐29 cells treated with various concentrations of COM. This cytotoxic effect was not observed when COM was used to treat normal intestinal epithelial cells (IEC‐6). Flow cytometry analysis of the cell cycle after treatment with COM clearly showed that COM inhibited cellular proliferation of HT‐29 cells via G1 phase arrest of the cell cycle. The apoptotic effect of COM on HT‐29 cells was confirmed by annexin V‐propidium iodide apoptosis test. An increase in the expression of p53, pro‐apoptotic factor Bax and a decrease in the expression of anti‐apoptotic factor Bcl‐2 were also observed in a dose‐dependent manner. COM also down regulated the expression of pro‐caspase‐3 and pro‐caspase‐9 in a dose dependent manner. These data suggest a possible mechanism whereby COM could induce apoptosis and cell cycle arrest. This property of COM also suggests that COM could have possible therapeutic potential against human colon adenocarcinoma.
Adult‐onset obesity induced by early life overnutrition could be reversed by caloric restriction
Authors: Liu Hung‐Wen , Mahmood Saleh , Srinivasan Malathi , Smiraglia Dominic J , Patel Mulchand
Published: June 2021

Overnutrition during the suckling period (small litter, SL) results in the development of adult‐onset obesity. The aim of the study was to investigate if caloric restriction (CR) in the postweaning period can reverse this phenomenon. The control litter (CL) had 12 pups/dam and SL had 3 male pups/dam from the postnatal day 3 until 21. In the postweaning period rats consumed lab chow as indicated: (i) CL and SL groups were on ad libitum regimen up to day 140, (ii) SL/CR group received 24% reduction in daily food intake compared to SL, and (iii) SL/CR/AL group was on 24% CR up to day 94 and then switched over to ad libitum feeding. CR caused reduction in body weight gains and normalized serum glucose, insulin, and leptin levels in SL/CR and SL/CR/AL rats compared to CL. The expression of neuropeptide Y (Npy) and leptin receptor (Lepr) returned to control levels in the hypothalami from SL/CR and SL/CR/AL rats compared to CL. No differences in the expression of pro‐opiomelanocortin and insulin receptor (Insr) were observed among these groups. Altered methylation of specific CpG dinucleotides in the proximal promoter region was observed for the Npy and Insr among these groups. CR resulted in normalized metabolic phenotype and gene expression involved in controlling food intake. The results show that the CR can be a successful intervention for reversing obese phenotype induced by lactational overfeeding.

Grant Funding Source: NIH DK61518
Cav1.3 does not contribute to active 1,25D‐regulated intestinal Ca absorption
Authors: Fernandez Perla Citlali Reyes , Replogle Rebecca , Li Qiang , Fleet James C
Published: June 2021

Intestinal Ca absorption occurs through a 1,25(OH)2D3 (1,25D)‐regulated transcellular pathway when habitual dietary Ca intake is low. Recently the L‐type Ca channel, Cav1.3, has been proposed to mediate active Ca absorption. We examined the role of Cav1.3 in Ca absorption using mouse models. Study 1 examined the relationship between Cav1.3 mRNA level and Ca absorption in a genetically diverse population of mice fed a normal (0.5%) or low (0.25%) Ca diet from 3–12 wks of age. Ca absorption was measured by oral gavage of a buffer containing 10 mM fructose and Ca45. Low Ca diets induced an increase in 1,25D and Ca absorption. Cav1.3 mRNA was detected in the duodenum, however, it was not associated with Ca absorption (r=−0.09, p=0.3) nor was it increased by low Ca intake (p=0.4). Study 2 examined whether glucose‐induced depolarization of the apical membrane of enterocytes enhances Ca absorption through Cav1.3. 10 wk‐old mice were fed high (1%) or low (0.125%) Ca diets for 1 wk. Ca absorption was examined using transport buffers containing Ca45 and either 25 mM glucose or fructose. While low Ca intake doubled Ca absorption, the sugar in the transport buffer did not increase Ca absorption efficiency (p=0.6) regardless of diet (p=0.19). Our data provide no support for the hypothesis that Cav1.3 and glucose‐induced depolarization are critical components of Ca absorption physiology.

Grant Funding Source: NIH awards DK54111 and ES019103 (JCF)
Early growth response‐1 in the pathogenesis of cysteamineinduced duodenal ulcer
Authors: Khomenko Tetyana , Deng Xiaoming , Patel Khushin N , Sandor Zsuzsanna , Szabo Sandor
Published: June 2021

We showed earlier that the immediate‐early gene & its zinc finger transcription factor early growth response (Egr) product were enhanced, including the expression of target genes (e.g., bFGF, PDGF, VEGF) in early stage of cysteamine (C)‐induced DU. Our recent studies demonstrated that deletion of egr‐1 gene led to 4.4‐fold increase in C‐induced DU in egr‐1 knockout (KO) vs. wild‐type (WT) mice. mRNA expression of bFGF, VEGF & PDGF was still increased but protein expression of bFGF, VEGF & PDGF was diminished in egr‐1 KO mice. We hypothesized that specific miRNAs in egr‐1 KO mice might be activated & inhibit protein synthesis of egr‐1 target genes. Using miRXplore Microarray study we found that 11 miRNAs (e.g., mir‐696, mir‐467E, mir‐152 & mir‐301B) were elevated by 1.8‐ to 1.9‐fold in egr‐1 KO mice vs. WT mice. Mir‐1892 decreased by 1.7‐fold in egr‐1 KO vs. WT mice. These miRNAs partially share validated targets relating to growth factor responses, e.g., FGFR3 & VEGF. We also demonstrated that DU was significantly reduced in rats treated with adenovirus encoding egr‐1 on days 7 & 14 (p < 0.05) respectively. The mRNA & protein expressions of egr‐1, bFGF, VEGF & PDGF markedly increased in rats after egr‐1 gene therapy. We conclude that Egr‐1 plays a preventive role in the pathogenesis of DU & further studies are needed to identify miRNA expression in egr‐1 KO & WT mice with DU.

(This work was supported by the Veterans Health Administration Merit Review Grant to S. Szabo).
Combined Foxp3 and IgG4 staining reveal distinct subsets of inflammatory bowel disease
Authors: Seidman Michael A , Bellizzi Andrew M
Published: June 2021

The two disease model in idiopathic inflammatory bowel disease (IBD), i.e. ulcerative colitis (UC) vs. Crohn's disease (CD), insufficiently captures the diseases' genetic and phenotypic diversity. We have utilized immunohistochemical staining for two markers of “regulatory” immune cells, IgG4 and FoxP3, to better characterize IBD, hoping to improve diagnosis, prognostication, and therapy. These stains were examined in cases of CD (n=11), UC (n=11), and demographically equivalent disease‐free patients (n=22). Biopsies taken from sites throughout the colon at all levels of disease activity were evaluated. The maximal number of cells staining per high power field (hpf) was scored. Neither IgG4 nor FoxP3 counts correlated with diagnosis (p>;0.05). Both correlated with disease activity (both p≈0.01). Neither showed correlation with age, gender, or anatomic site. Generalized linear models did not determine IgG4 or FoxP3 counts to be good predictors of diagnosis, even when corrected for disease activity. However, supervised hierarchical clustering revealed that the combination of the two stains identified clinically meaningful subsets of disease (Figure 1).

These data provide insight into IBD that will help guide future understanding and therapies, going beyond the currently prevailing two disease paradigm.

Research was supported entirely by internal (i.e. departmental) funds.
Identification and assessment of regulatory T cells (Treg*) associated with inflammation and tumorigenesis in Inflammatory Bowel Disease (IBD) patients
Authors: Downey Katherine J , Wang Yanhua , Sun Katherine , Lin Elain , Wu Yuanyuan , Liu Qiang
Published: June 2021

The pathogenesis of Inflammatory Bowel Disease is multifactorial. A widely accepted theory for the pathogenesis of IBD lays blame on the host's exaggerated immune response to commensal bacteria, implying a loss of self tolerance. Chronic inflammation is a well known risk factor for the development of dysplasia and cancer. Regulatory T cells (Treg*) therefore play a critical role in the regulation of immunity and a pivotal role in tumorigenesis of several cancer types by suppressing anti‐tumor immune responses, thereby promoting tumor progression. We set out to identify and assess Treg* expression in IBD and Cancer patients vs. disease‐free individuals which may help explain tumor progression.

Retrospective sequential in‐house cases (n=53): 34 IBD, 9 Colon cancer, and 10 normal. Immunohistochemistry used to delineate Treg* cells. Blinded counts performed using the average of 3 HPF's (40X) in areas of highest concentration. Data analysis by unpaired t‐test and ANOVA reveal significant differences in Treg* expression: IBD >; normal (p<0.001); Cancer >; normal (p<0.001 ); Cancer >; IBD (p<0.003); no significant difference between UC and Crohn's (p=0.371). ANOVA analysis showed statistically significant difference amongst all groups (p<0.001: Cancer >; IBD >; normal).

Our results show increased Treg* cells in the cancer arm (cancer >; IBD >;>; normal), thereby supporting an important role of Treg* cells in inflammation and tumorigenesis in IBD. Therapeutic manipulation of Treg* cells (i.e. selective depletion) holds great potential in treating IBD and preventing subsequent tumor progression.

Funding provided by Montefiore Medical Center Clinical Research Committee
Pharmacological inhibition of p38 mitogen‐activated protein kinases affects KC/CXCL1‐induced intraluminal crawling, transendothelial migration, and chemotaxis of neutrophils in vivo
Authors: Liu Lixin , Hossain Mokarram , Qadri Syed M
Published: June 2021

Leukocyte recruitment to the inflammation site is a multi‐step process governed by specific signalling cascades. After adhesion in the lumen, leukocytes crawl to optimal sites at endothelial junctions and transmigrate to extravascular tissue in a Mac‐1‐dependent manner. The signalling mechanisms that regulate post‐adhesion steps of intraluminal crawling, transmigration and chemotaxis in tissue remain incompletely understood. The present study explored the effect of p38 MAPK inhibitor SB203580 on various steps of neutrophil recruitment triggered by chemokine KC (CXCL1) gradient in microvasculature of murine cremaster muscle using intravital microscopy and time‐lapsed video analysis. SB203580 (100 nM) did not change leukocyte rolling but significantly attenuated neutrophil adhesion, emigration, transmigration time and detachment time, and impaired the initiation of crawling and transmigration. In response to KC chemotactic gradient, SB203580 significantly reduced the velocity of migration and chemotaxis index of neutrophils in tissue. The upregulation of Mac‐1 expression in neutrophils stimulated by KC was significantly blunted by SB203580 in vitro. Collectively, our findings demonstrate that pharmacological suppression of p38 MAPK significantly impairs multiple steps of neutrophil recruitment in vivo.
Kinin B1 receptor regulates inflammatory cell recruitment during low laminar shear stress
Authors: Martin Katherine , Perretti Mauro , Ahluwalia Amrita
Published: June 2021

The kinin B1 receptor (B1R) is an inducible receptor that is only weakly expressed, if at all, under normal physiological conditions but is induced under situations of stress. We have previously shown that when endothelial cells are exposed to inflammatory stimuli or low laminar shear stress (LSS), two factors commonly associated with atherosclerosis, B1R expression is induced. In vivo, B1R agonists stimulate inflammatory cell recruitment including neutrophils and macrophages. We now show in vitro that subjecting human aortic endothelial cells to low LSS enhances leukocyte adherence compared to physiological LSS. Moreover, this effect is enhanced in the presence of the cytokine interleukin‐1β, both conditions associated with increased endothelial B1R expression. Importantly treatment of endothelial cells with a B1R antagonist reduced cell adherence in cells exposed to low LSS but had no significant effect on the response under physiological LSS conditions. These results suggest that B1R may play an important role in inflammatory cell recruitment at sites of low LSS and targeting this protein might prove beneficial in the treatment of atherosclerosis.

This work and KRM are funded by The British Heart Foundation
Role of Toll‐Like Receptor 4 in Legionella pneumophila infection with C3H/HeJ Mouse Model
Authors: Wu Wei‐wei , Wang Su‐ping , Zhang Fan , Gao Xiang‐dong , Dong Jieming
Published: June 2021

Objective

To explore the relationship between TLR4 and Legionella infection in a mouse model.

Methods

C3H/HeJ and C3H/HeN mice were used as experimentl animal. Infection group and contol group were tracheal injection of bacterial suspension or sterile saline nasal respectively. All mice were sacrificed at 12h, 24h, 48h, 72h respectively. Left lung upper lobe was taken to do fixed biopsy. TLR4 protein expression level of each group on PBMC were detected by flow cytometry.

Results

Pathological examination showed that the C3H/HeJ and C3H/HeN infection group in lung tissue were showed infiltration of inflammatory cells and alveolar vascular bleeding, the control group was not found inflammatory reaction. Legionella pneumonia model established successfully. There was significantly difference of TLR4 protein expression in three groups, each timing and the interaction of groups and timing(P<0.001). Comparison between groups found that C3H/HeJ infection group and C3H/HeN infection group, C3H/HeJ infection group and C3H/HeN control group shows statistical significance(P<0.001). But C3H/HeN infection group and C3H/HeN control group had no statistical significance(P>;0.05).

Conclusion

The TLR4 gene mutations make it low reaction to Legionella. However, the anti‐inflammatory effects of TLR4 to Legionella needs further study.

This study was supported by the National Natural Science Funds of China (grant no. 30872154)
Group B Streptococcus Induces a Caspase‐Dependent Apoptosis in Fetal Rat Lung Interstitium
Authors: Kling David Eric , Murphy Miriam P , Newburg David S
Published: June 2021

Group B Streptococcus (GBS) is an important fetal, neonatal, and infant pathogen. Using an ex vivo infection model, GBS induced profound reductions in fetal lung; explant size, airway branching, and erythroblast island areas. Elevated levels of apoptosis subsequent to GBS infections were observed by whole‐mount confocal immunofluorescence using activated‐caspase‐3‐antibodies and terminal deoxynucleotidyl transferase dUTP nick end‐labeling (TUNEL) assays. The caspase inhibitor Z‐VAD‐FMK abolished the increase in TUNEL‐positive cells associated with GBS infections, indicating that the GBS‐induced apoptosis was caspase‐dependent. Digital image analyses revealed that both GBS and the active form of caspase‐3 were distributed primarily within the lung interstitium, suggesting that these tissues are important targets for GBS. Antibodies to the active form of caspase‐3 colocalized with both macrophage‐and erythroblast‐markers, suggesting that these hematopoietic cells are GBS targets. These studies suggest that GBS infections profoundly alter lung morphology and that caspase‐dependent hematopoietic cell apoptosis within the lung interstitium.
Identifying Mechanisms of Interleukin‐19‐Mediated HuR‐Dependent Reduction in Vascular Cell mRNA Stability
Authors: England Ross Nevin , Kelemen Sheri , Ellison Stephen , Autieri Michael
Published: June 2021

IL‐19 reduces TNF‐α‐driven mRNA abundance and protein abundance of intercellular adhesion molecule‐1 (ICAM‐1) and vascular cell adhesion molecule‐1 (VCAM‐1) (p<0.01) in cultured coronary artery endothelial cells (ECs) as measured by RT‐PCR and immunoblot. IL‐19 treatment produced no change in activation of the transcription factor NF‐κB, as measured by immunoblot analysis of p65 phosphorylation, translocation, and IkB degradation. IL‐19 decreases stability of ICAM‐1 and VCAM‐1 mRNA in actinomycin‐D treated cells. IL‐19 treatment inhibits cytoplasmic translocation of the mRNA stability factor HuR as assayed by immunoblot and immunofluorescence. siRNA knock‐down of HuR also decreases ICAM‐1 and VCAM‐1 mRNA stability. IL‐19 decreases HuR cytoplasmic translocation and phosphorylation as measured by phospho‐serine immunoprecipitation of HuR. In cultured coronary artery smooth muscle cells (SMCs) and ECs, IL‐19 increases expression of putative HuR regulator microRNA‐133, measured using the miScript PCR system (Qiagen), and treatment with miR‐133 mimic reduced expression of HuR mRNA.

These data support our hypothesis that IL‐19 exerts its anti‐inflammatory effects by HuR‐mediated decrease in mRNA stability. We propose inhibition of HuR phosphorylation and translocation and upregulation of miR‐133 as possible mechanisms.
Modulation of the receptor ALX/FPR2 in vascular inflammation
Authors: Perretti Mauro , Bena Stefania , Brancaleone Vincenzo , Flower Roderick J
Published: June 2021

The receptor ALX/FPR2 conveys the tissue protective properties of lipoxin A4 and Annexin A1 (AnxA1). Here, we tested if ALX was modulated during ischemia/reperfusion (IR) to attenuate secondary organ injury. A mouse colony nullified for ALX and with a GFP reporter was used. Acute myocardial infarct (25/120 min IR) was characterised by high mortality (~40%) in Alx null, but not wild type, mice. Using 90 min reperfusion revealed a higher degree of heart necrosis in the null mouse. In the lung, Alx promoter activity was evident in resting conditions, with signal in bronchial epithelia and scattered macrophages, and was markedly upregulated after IR (qPCR and immunohistochemistry). Higher KC, TNFα and IL6 levels were quantified in lung extracts of Alx null mice and were associated with augmented MPO activity values. These data indicate profound modulatory functions of ALX/FPR2 in IR with rapid up‐regulation and non‐reduntant functions in distant organs such as the lung.

Supported by The Wellcome Trust
Differential expression of pro‐inflammatory and pro‐resolving lipid signaling genes in ARDS
Authors: Abdulnour Raja‐Elie Edward , Howrylak Judie A , Dolinay Tamas , Choi Augustine , Levy Bruce D
Published: June 2021

RATIONALE

Lipid mediators play important roles in the pathophysiology of the acute respiratory distress syndrome (ARDS), so we investigated the differential expression of genes associated with lipid signaling in patients with ARDS.

METHODS

For patients that met clinical criteria for sepsis with ARDS (n= 19) and sepsis alone (n=29) whole blood was obtained on admission to the medical intensive care unit (day 0) and on day 7. RNA was extracted and microarray analysis performed to compare the expression profile of genes related to eicosanoid biosynthesis, function, and regulation between the two groups. Validation of select candidate genes was performed in experimental ARDS in BALB/c mice (50μl 0.1N HCl, pH 1.0 or saline) by semi‐quantitative PCR on lung RNA.

RESULTS

Several pro‐inflammatory lipid genes were upregulated at ARDS day 0 and downregulated at day 7 including PLA2G2A, ALOX5, ALOX5AP, PTGS2, PTGER3, and LTB4R. In contrast, genes linked to resolution displayed distinct patterns of expression; ALOX15, ALOX15B and PTGER4 were downregulated whereas AHR, GPR32 and ALX/FPR2 were upregulated. Genes involved in phosphatidic acid, sphingomyelin, and polyisoprenyl diphosphate metabolism were differentially regulated. For example, PLD1 and SGMS2 were upregulated at day 0 and downregulated at day 7, while PPAP2C and PPAPDC2 were upregulated at both time points. Analysis of gene expression in a murine model of acute lung injury confirmed selected target genes.

CONCLUSION

These findings emphasize that pro‐inflammatory and pro‐resolving lipid signaling genes are subject to temporally distinct regulation in the early phases of ARDS, and have uncovered several new potential points of regulation. The work is supported in part by the NIH (T32‐HL07633, R01‐HL068669, P01‐GM095467)
Identification of differentially expressed microRNAs in rat brain neuronal culture induced by Angiotensin II
Authors: Singh Neha , Guo Lirong , Zhong Tiecheng , O'Rourke Stephen T. , Sun Chengwen
Published: June 2021

The brain renin–angiotensin system (RAS), particularly Angiotensin II (Ang II) plays an important role in the central control of blood pressure. However, the role of microRNAs (miRs) in this process is unclear. Our aim was to identify the miRs that are altered by Ang II treatment in rat brainstem and hypothalamus neurons. A genome wide miRNA array Quantitative PCR was performed and analyzed in neuronal cultures treated with or without Ang II (100 nM, 24h). Analysis of miRNA arrays revealed several miR levels were altered by Ang II (100 nM, 24h) as compared to control neurons. Five miRs that were most up‐regulated by Ang II were miR‐219–1‐3p (18.4 fold), miR‐466c (15.7 fold), miR‐136 (10.2 fold), miR‐27a* (10.1 fold), miR‐25* (10 fold); and those that were most down‐regulated were miR‐9* (139 fold), miR‐20b‐5p (89.5 fold), miR‐20a (83.6 fold), miR‐26a (74.3fold) and miR‐124 (74.1 fold). Several critical genes have been identified by bioinformatic analysis for the validated target genes of these miRs, such as cannabinoid receptors (CB1R), Neuregulin 1 and STK39, suggesting that Ang II might be acting through these differentially expressed miRs to regulate other gene expression. In this study, we identified several miRs altered by Ang II treatment, which may shift neuronal gene profile, contributing to the long‐term blood pressure regulation.

Research Support: NDSU‐COBRE, ND‐EPSCoR
In vivo Evans blue dye infusion allows monitoring of auto‐reperfusion in a rat model of cardiac ischemia
Authors: Liao Kate Hsiurong , Kuo Eva Yuhua , Chen Kuen‐Bao , Lai Ted Weita
Published: June 2021

One major problem with animal models of cardiac ischemia is the huge variability in the degree and the actual duration of coronary occlusion, in large because of auto‐reperfusion that results in early‐onset partial‐reperfusion and the subsequent partial‐ischemia. The Evans blue dye (EBD) is a commonly used tracer for defining the boundary of ischemia in excised heart with re‐occluded coronary artery. This same dye is also commonly used in vivo for blood‐brain barrier research, and has been used safely in human subjects. In this study, we intravenously injected the rat with EBD immediately following left coronary arterial ligation in vivo, and observed its initial clear‐cut exclusion from the ischemic region. This initial exclusion was later followed by slow leakage of EBD into the ischemic region, at a pace depending on the “tightness” of the initial occlusion. Moreover, complete reperfusion of the heart resulted in the accumulation of EBD in the ischemic region, after which perfusion of the excised heart resulted in a visible clear‐cut boundary showing EBD‐filled ischemic region, which closely resembles the initial pre‐marked ischemic region. In conclusion, our refined rat model of cardiac ischemia allows for in vivo detection of early auto‐reperfusion and post hoc ex vivo detection of initial area of ischemia, and thus can effectively reduce technical variability in pre‐clinical experiments. (NSC of Taiwan)
Interplay between complement factor B and Toll‐like receptors and its role in septic cardiomyopathy
Authors: Zou Lin , Li Yan , Feng Yan , Zhang Ming , Chen Chan , Chao Wei
Published: June 2021

Objective

The goal of the current study was to determine whether complement factor B (cfB) would interplay with Toll‐like receptors (TLRs) during sepsis and whether or not such interplay would contribute to septic cardiomyopathy.

Methods

Cardiomyocytes (CMs) were isolated from wild‐type (WT) mice or mice deficient of TLR2 or TLR4. Polymicrobial sepsis was created by cecum ligation and puncture (CLP). cfB mRNA and protein were determined by qRT‐PCR and Western blot, respectively. Cytokines were determined using a fluorescent multiplex assay. Left ventricular function was assessed in a Langendorff perfusion system.

Results

Both Pam3cys and lipopolysaccharide (LPS) markedly increased cfB mRNA expression in CMs via TLR2‐ and TLR4‐dependent manner (fold, 217±54 and 578±54, respectively), but not complement 3, 4 or 5. TLR2, TLR3, TLR4 ligands also induced marked cfB protein expression in and release from CMs. In vivo, CLP led to a marked and time‐dependent increase in cfB mRNA and protein expression in septic heart via MyD88‐dependent manner. Importantly, compared with septic WT mice, septic cfB‐KO mice had improved LV function, better survival rate, and decreased cytokine production.

Conclusion

cfB is up‐regulated in response to TLR activation and polymicrobial sepsis via TLR‐dependent manner. cfB deficiency confers a beneficial effect in cardiac function during sepsis.

This research has been supported by NIH R01‐GM080906 and R01‐GM097259.
Angiotensin II receptor blockade, but not ACE inhibition, reduces nocturnal hypertesion and natriuresis in autonomic failure patients with low renin activity
Authors: Arnold Amy C , Okamoto Luis E , Gamboa Alfredo , Shibao Cyndya A , Raj Satish R , Robertson David , Biaggioni Italo
Published: June 2021

At least half of autonomic failure [AF] patients exhibit paradoxic supine hypertension. However, the underlying mechanisms and optimal treatment strategies for this condition are unknown. While renin activity is often undetectable, we recently showed that circulating Ang II levels are elevated in AF patients. Thus, we tested the hypothesis that endogenous Ang II contributes to the hypertension of AF. The AT1 receptor blocker losartan (50 mg) was administered at bedtime to 11 hypertensive AF patients in a randomized, double‐blind, placebo‐controlled study. Seven AF patients also received the ACE inhibitor captopril. Plasma Ang II was increased in AF patients [52±5 pg/ml] relative to matched healthy subjects [27±4 pg/ml; p=0.002], despite similar low renin activity. Losartan maximally reduced systolic blood pressure by 32±11 mmHg [p<0.05], decreased nocturnal urinary sodium excretion [0.111±0.015 losartan vs 0.151±0.008 mmol/mg placebo, p=0.046] and did not worsen morning orthostatic tolerance [404±163 losartan vs 530±194 mmHg*min placebo, p=0.687]. There was no effect of captopril on supine blood pressure, sodium excretion or orthostatic tolerance in AF perhaps suggesting that the Ang II generation is independent of ACE activity. These findings suggest that elevations in Ang II contribute to the hypertension of AF and provide new rationale for use of AT1 receptor blockers in treatment of these patients. Support: NIH grants P01 HL‐056693, UL1 TR000445 and AHA 11POST7330010.