bluuebunny/crossref_metadata_2025 huggingface dataset:
Mice fed vitamin A‐modified diets develop distinct levels of tissue retinol depending on growth stage and vitamin A exposure (645.23)
Authors: Wei Cheng‐Hsin , Chen Qiuyan , Ross A.
Published: June 2021

The objective of this study was to determine the effects of growth stage (age) and dietary vitamin A (VA) on retinoid levels in plasma and main retinoid storage organs in a mouse model that is commonly used for immunological research. A purified VA‐marginal diet was fed to Balb/C dams from pregnancy until pups were weaned. Post‐weaning pups (total n=46) were randomized to 4 diet groups and fed VA‐deficient, marginal, adequate, and supplement diets, which contains 0, 0.35, 4, and 100 μg retinol as retinyl palmitate/g diet, respectively. Tissues were collected from newborn (0 d), neonatal (14 d), young adult (4‐wk), and adult (8‐wk) mice and analyzed for total retinol (ROH) by UPLC. A dose‐response relationship was observed in liver and lung, but not plasma. Surprisingly, the liver and lung ROH levels in VA‐deficient and marginal groups for young adult and adult mice were higher as compared to those in rats previously studied in our laboratory. The results suggested that under VA‐adequate and supplemented conditions mice are capable of accumulating high levels of VA in tissues, especially in lung, with some accumulation even on a VA‐marginal diet. Additionally, there were small but significant differences in the percentage of immune cell populations in lung and spleen between VA‐deficient and supplemented mice. Understanding VA physiology in the growing mouse will be helpful in making the best use of this common animal model.

Grant Funding Source: NIH DK‐41479
Effect of a 2‐week intense life‐style intervention followed by 6‐month carotenoid supplementation on fat depots, aditonectin and palmitoleate: a 6‐month double blind placebo‐controlled pilot study in obese children (645.6)
Authors: Canas Jose , Lochrie Amanda , Galena McGowan Amy , Schettino Christopher , Hossain Jobayer , Balagopal Prabhakaran
Published: June 2021

Background and Objectives: Abdominal subcutaneous and visceral adipose tissues (SAT and VAT) contribute to the established association between obesity and metabolic risk, likely through signaling molecules that regulate insulin resistance (IR). High molecular weight adiponectin (HMW‐ADI) and C16:1n7‐palmitoleate (PAL) play crucial roles in this respect. Carotenoids have emerged as possible adiposity ameliorating agents, but their role in the regulation of these signals and their impact on fat depots remain less clear. Method and Design: Of the 21 obese children recruited, 17 (Age = 10.5 ± 5 years) completed a randomized double blind placebo controlled study of a 2‐week intense lifestyle intervention program (LSIP) followed by 6 months of carotenoid supplementation (Jarrow Formulas CaroteneAll®). β‐carotene, HMW‐ADI, glucose, insulin and PAL were measured. HOMA‐IR was calculated and the percent change in VAT and SAT were determined by cross‐sectional multi‐slice MRI (1.5‐T; GE Medical Systems) and DEXA (QDR‐4500W; Hologic) at baseline and 6 mo.

Results: β‐carotene was inversely related to BMI‐Z; VAT and SAT (p<0.05 for all). Supplementation resulted in 95 ± 66% increase in β‐carotene at 6 months as compared to 3 ± 19% decrease in the placebo group (p=0.001). HMW‐ADI increased by 43% ± 88% (p<0.03) with the supplement and decreased by 5.6 ± 37% (p = NS) in the placebo group. PAL was lowered by 16 ± 36% at 6 months (p = 0.03) in the supplement group. There were no significant changes in triglycerides or HOMA‐IR. There was a 4% decrease in mean SAT by MRI and an 8% decrease in mean estimated VAT by DEXA (p< 0.05 for all) in the supplement group.

Conclusion: The study results showing an increase in HMW‐ADI and a decrease in PAL along with beneficial changes in fat depots in response to the 2‐week LSIP followed by carotenoid supplementation are novel in obese children. The mechanistic role of these divergent changes warrants further investigation.
Iron biofortified beans improve iron status in Rwandan University women: results of a feeding trial (646.1)
Authors: Haas Jere , Luna Sarah , Lung'aho Mercy , Ngabo Fidel , Wenger Michael , Murray‐Kolb Laura , Beebe Steve , Gahutu Jean‐Bosco , Egli Ines
Published: June 2021

Objective: To examine the efficacy of an iron biofortified bean intervention on iron status in iron depleted young women.

Methods: 257 Rwandan university women (18‐27 y) with screening ferritin <20 μg/L were randomized to consume ~155 g/d, dry wt. of either iron biofortified beans (Fe=78 mg/g) or control beans (Fe=59 mg/g) per day for 135 days. Iron status (hemoglobin‐Hb, ferritin‐Fer, transferrin receptor‐TfR, body iron‐BI), inflammation (CRP, AGP), and anthropometric indices were determined at enrollment, 7 random midpoints and after 135 days. Difference‐in‐difference analysis plus repeated measures ANOVA and linear and binomial regression models were used to evaluate the effect of the intervention on iron status.

Results: At baseline, for the 231 study subjects who completed the study, 34% were anemic (Hb <120 g/L), 73% had Fer <15.0 μg/L) and 48% had negative BI stores (Cook’s algorithm). After 135 days of feeding, there was significant group difference in change in Hb (difference=3.5 g/L, p<.001) and BI (difference=0.45 mg/kg, p=.033) in the high iron compared to control group. Also, 54 of 109 women with negative BI at baseline increased their BI by 1.5 mg/kg, with 30% more responders in the high iron compared to the control group.

Conclusions: Consumption of iron‐biofortified beans over 4.5 months improved iron status in iron depleted Rwandan women.

Grant Funding Source: HarvestPlus/IFPRI
Bifidobacterium bifidum R0071 decreases stress‐associated diarrhea‐related symptoms and ultimately self‐reported stress in a randomized, double‐blind, placebo‐controlled study (647.21)
Authors: Nieves Carmelo , Rowe Cassie , Culpepper Tyler , Christman Mary , Langkamp‐Henken Bobbi
Published: June 2021

Psychological stress is associated with gastrointestinal (GI) distress. This study determined whether probiotics could normalize stress‐associated GI distress and ultimately reduce overall stress. Undergraduate students (20±2 y) undergoing academic stress (final exams at wks 4 to 5) consumed a probiotic (Lactobacillus helveticus R0052, Bifidobacterium bifidum R0071, or Bifidobacterium longum ssp. infantis R0033) or a placebo (n=142 to 147/grp) and self‐reported stress (0=no stress to 10=extremely stressed) daily for 6 wks. Each wk participants scored 4 diarrhea‐related symptoms (DS, 1=no discomfort to 7=severe discomfort) using the GI Symptom Response Scale; scores were summed. Stress was positively related to DS (P=0.0562). DS were lower with B. bifidum vs placebo at wks 2 and 5 and vs L. helveticus at wk 5. There was an interaction between the intervention and antibiotic use in the prior wk where DS were higher with antibiotics with L. helveticus (P=0.056) and placebo (P=0.009). There was no difference in DS with or without antibiotic use with B. bifidum or B. longum. Only B. bifidum had an effect on stress scores (P=0.009) which was also dependent on hours of sleep where the stress score decreased by 0.13 for each additional hour of sleep. These data suggest that during a stressful period B. bifidum decreases DS, even in the presence of antibiotics, and ultimately decreases self‐reported stress scores.

Grant Funding Source: Lallemand Health Solutions
Kefir exhibits anti‐proliferative and pro‐apoptotic effects on colon adenocarcinoma cells with no significant effects on cell migration and invasion (647.24)
Authors: Rizk Sandra , El Khoury Nathalie , El‐Hayek Stephany , Tarras Omayr , El‐sibai Mirvat , El‐Sabban Marwan
Published: June 2021

Background: Previous studies showed that kefir, a fermented milk product, exhibited anti‐tumoral activity in vivo; yet its mechanism of action remains elusive. Recent studies have focused on the mechanism of action of kefir on cancer cells in vitro. The current study aims at examining the effect of kefir on cell survival, proliferation and motility of colorectal cancer (CRC) cells.

Methods: Kefir’s anti‐cancer potential was tested on CRC cell lines, Caco‐2 and HT‐29, through cytotoxicity, proliferation, and apoptotic assays. The expression of certain genes involved in proliferation and apoptosis was assessed using RT‐PCR and western blot analysis to determine the possible mechanism of action of kefir. To assess the effect of kefir on cancer metastasis, wound healing and time‐lapse movies, in addition to collagen‐based invasion assay, were used.

Results: Results have shown that cell‐free fractions of kefir exhibit an anti‐proliferative effect on Caco‐2 and HT‐29 cells. Analysis of DNA content by flow cytometry revealed the ability of kefir to induce cell cycle arrest at the G1 phase. Kefir was also found to induce apoptosis, as seen by cell death ELISA. Results from RT‐PCR showed that kefir decreases the expression of TGF‐α and TGF‐β1 in HT‐29 cells. Western blot results revealed an upregulation in Bax:Bcl‐2 ratio, confirming the pro‐apoptotic effect of kefir, and an increase in p53 independent‐p21 expression upon kefir treatment. MMP expression was not altered by kefir treatment. Furthermore, results from time‐lapse motility movies, wound‐healing, and invasion assays showed no effect on the motility of colorectal as well as breast (MCF‐7 and MB‐MDA‐231) cancer cells upon kefir treatment.

Conclusion: Our data suggest that kefir is able to inhibit the proliferation and induce apoptosis in HT‐29 and Caco‐2 CRC cells, yet it does not exhibit a significant effect on the motility and invasion of these cells in vitro.

Grant Funding Source: Supported by SRDC, Lebanese American University
Effect of probiotic BIO‐L6® on intestinal morphology, microbiota and serum cytokines in obese Zucker rats (647.33)
Authors: Torres Aguilar Lorena , Reyes Esparza Jorge , Rodríguez Fragoso Lourdes
Published: June 2021

The aim of this study is to investigate the effects of probiótico BIO‐L6® (a mixture of 10 strain of lactobacilli) on intestinal microbiota (lactobacillus and coliforms levels) as well as seric cytokines (TNF, IL‐6, IL‐10 and IL‐4) in Zucker rats. Intestinal microbiota plays a role in the development of obesity and its associated low‐grade inflammation; probiotics may be useful in preventing and/or improving obesity. Twenty male lean and obese (fa/fa) Zucker rats were assigned in four groups and administered v.o with BIO‐L6® (8.8x106colony‐forming units/rat) daily for 9 weeks. Fecal lactobacillus and coliforms levels and serum cytokines (TNF, IL‐6, IL‐10 and IL‐4) were measured. Intestinal stain of H&E was evaluated. BIO‐L6® treatment decreased total coliforms, 4.5 times in lean rats and 90% in obese rats (p<0.05), and increased 82% and 48% the lactobacillus levels (p<0.05). With respect with the obese groups, BIO‐L6® treatment decreased TNF 19.6% and increased IL‐4 and IL‐10 (35% and 53.8%). In the bowel histology was observed an increase in the number of goblet cells in both groups treated. In conclusion, the BIO‐L6® probiotic had anti‐inflammatory effect in rats Zucker, moreover, altered intestinal microbiota because increased lactobacilli and decreased coliforms and demonstrated an increase in the production of cells responsible for the secretion of intestinal mucus.
Failure of recovery from lead‐acetate‐induced oxidative stress and cardiotoxicity in Wistar rats (648.14)
Authors: Omobowale Temidayo , Ademola Oyagbemi , Saba Adebowale , Akinrinde Stephen , Daramola Temilade , Ogunpolu Blessing
Published: June 2021

Lead poisoning continues to be a serious health problem and more significantly so in developing countries with ineffective waste disposal systems. Recent efforts at solving lead poisoning issues have seen entire towns being re‐settled from lead‐contaminated areas. This study was designed to investigate whether withdrawal of lead exposure results in a resolution of toxic effects of lead to cardiac tissues. Adult male Wistar rats were exposed orally to lead acetate at doses of 0.25, 0.5 and 1.0mg/ml for 6‐week duration, after which one‐half was sacrificed and the remaining left for a further 6 weeks without lead treatment. Exposure of rats to lead acetate produced significant decline (p<0.05) in the activities of antioxidant parameters, including Glutathione peroxidase (GPx), Glutathione S‐transferase (GST), Catalase (CAT), Superoxide dismutase (SOD) and Reduced glutathione (GSH), while Malondialdehyde (MDA) concentration was significantly elevated. Animals from the withdrawal period exhibited similar pattern of alterations, with significant (p<0.05) reduction in GSH, GPx and SOD and significant elevation in MDA and H2O2 concentrations. However, GST activity was elevated, while CAT activity remained unaltered in the recovery period. The results of this study showed that cardiotoxicity indicated by induction of oxidative stress and reduction in antioxidant parameters in this study failed to resolve upon withdrawal of lead exposure in male rats during the period of study.
Novel insights in post‐infectious irritable bowel syndrome in experimental giardiasis (650.15)
Authors: Halliez Marie , Le Goff Laetitia , Francois Arnaud , Colasse Elodie , Gargala Gilles , Villena Isabelle , Favennec Loic , Lapointe Tamia , Altier Christophe , Buret Andre
Published: June 2021

Giardia duodenalis infections can lead to post‐infectious irritable bowel syndrome (PI‐IBS), a disorder characterized by chronic visceral pain. Using a new model of post‐giardiasis IBS, this study aimed to reveal novel mechanisms responsible for PI‐IBS. Weaning rats were gavaged with live Giardia trophozoites (assemblages A or B). Visceral hypersensitivity was assessed 50 days post‐infection (DPI) by measuring blood pressure changes in response to jejunal/rectal balloon distension. Spinal cord expression of c‐fos was used as a marker of nociceptive signaling. Caco‐2 monolayers were used to assess non‐invasive E. Coli HB101 translocation, as well as the integrity of epithelial barrier structure and function at early time points. c‐fos activation was observed during the acute phase. While the infection was cleared by 21DPI, jejunal and rectal hypersensitivity was observed 50 DPI with either assemblage along with mast cell infiltration. In vitro challenge with Giardia also facilitated the paracellular translocation of E. coli, increased epithelial permeability, and disrupted the tight junctional (TJ) proteins occludin and claudin‐4. These data show that Giardia‐induced IBS is associated with TJ proteins disruption, translocation of a commensal E. coli and c‐fos activation during the acute phase of the infection, as well as mast cell infiltration and visceral hypersensitivity. Findings in this model will help identify the mechanisms of Giardia leading to post‐giardiasis‐IBS.

Grant Funding Source: Supported by the French ministry of secondary education and research, NSERC‐CREATE
Sensitization of sodium channels by cystathionine β‐synthetase activation in colon primary sensory neurons contributes to visceral hypersensitivity in adult rats with neonatal maternal deprivation (651.20)
Authors: Xu Guangyin
Published: June 2021

Aims: This study was designed to determine roles for sodium channels and endogenous H2S signaling in a rat model of chronic visceral hypersensitivityinduced by neonatal maternal deprivation (NMD). Methods: Neonatal male rats were exposed to a 3 hour period of daily maternal separation. Colon‐specific DRG neurons were acutely dissociated for measuring excitability and sodium channel currents under whole‐cell patch clamp configurations. Expression of NaV1.8 was analyzed by western blot. Results: NMD significantly increased AWR scores. TTX‐resistant sodium current density was greatly enhanced in colon DRG neurons, activation curves showed a leftward shift in NMD rats. Furthermore, NMD remarkably enhanced expression of NaV1.8 at protein levels. AOAA, an inhibitor of CBS, significantly suppressed the expression of NaV1.8. AOAA treatment also reduced the TTX‐R sodium current density, and reversed hyperexcitability of colon‐specific DRG neurons in NMD rats. Conversely, addition of NaHS, a donor of H2S, greatly enhanced TTX‐R sodium current density, leftshifted activation curve and enhanced excitability of colon DRG neurons in age‐matched healthy rats. Conclusion: Our data suggest that NMD enhances TTX‐resistant sodium activity of colon DRG neurons, which is most likely mediated by CBS‐H2S signaling, thus contributing to chronic visceral hypersensitivity.

Grant Funding Source: Supported by National Natural Science Foundation of China
UM206, a frizzled‐receptor antagonist attenuates adverse remodeling and cardiac function deterioration following myocardial infarction (652.10)
Authors: Hermans Kevin , Daskalopoulos Evangelos , Janssen Ben , Blankesteijn W. Matthijs
Published: June 2021

Activation of the Wnt/frizzled signaling pathway has a key role in wound healing and cardiac remodeling following myocardial infarction (MI). We have previously shown that Wnt/frizzled signaling blockade with UM206 immediately after MI suppresses adverse remodeling and prevents heart failure (HF) development in a mouse MI model. Here, we investigated the role of UM206 after short‐time early treatment and long‐term late treatment, in order to unveil the optimal anti‐HF regimen.

Swiss male mice were subjected to MI by left anterior descending coronary artery permanent ligation. Two different regimens were followed: a) short‐time early treatment: 2 weeks of UM206 treatment followed by 3 weeks without treatment and b) long‐term late treatment: 3 weeks without treatment followed by 5 weeks of UM206 treatment. UM206 was administered via an osmotic mini‐pump (6µg/kg/day) and controls were administered normal saline.

Early UM206 treatment led to improved end diastolic volume (178±6 mm3) compared to saline (214±20 mm3, p<0.01) or to late UM206 treatment (278±24 mm3, p<0.01). Both effects were less pronounced compared to the full UM206 treatment (132±30 mm3). Ejection fraction was improved in early (24.4±0.1%) and late (23.2±2.6%) treatment compared to saline (16.5±0.2, p<0.01 and 11.0±3.2%, p<0.05 respectively) but full treatment was the optimal (31.4±0.4%), p<0.001).

Both short‐time early and long‐term late UM206‐treatment regimens demonstrate a cardioprotective effect by halting the progression to HF. Nevertheless, full UM206 treatment immediately after MI is the one eliciting the most beneficial effects for the injured myocardium.

Grant Funding Source: Supported by the Dutch Heart foundation (2010B196) and Cyttron (FES0908)
The role of the RNA binding proteins HuR and AUF1 in β1‐adrenergic receptor mRNA stability in heart failure (652.13)
Authors: Kraynik Stephen , Anthony Sarah , Kelley Melissa , Tranter Michael
Published: June 2021

Cardiovascular disease is the leading cause of death in the United States. It is known that heart failure is marked by a decrease in expression of key proteins that regulate cardiac contractility, including the β1‐adrenergic receptor (β1‐AR). Despite a compensatory increase in sympathetic tone, cardiac contractility and inotropic response are substantially impaired, primarily through β1‐AR desensitization and mRNA degradation. Here we aim to elucidate the mechanisms by which the β1‐AR is regulated via mRNA processing at the regulatory 3’‐end (alternative polyadenylation), and identify the role that RNA binding proteins, such as HuR and AUF‐1, play in β1‐AR mRNA stability. We demonstrate a decrease in β1‐AR mRNA in cardiac pathology models of pressure overload induced heart failure (transverse aortic constriction; TAC) and chronic (2 week) β AR stimulation. In addition, we show the effect that cardiac‐specific deletion of HuR has on heart function and β1‐AR mRNA regulation. We also investigate the mechanisms of agonist‐dependent β1‐AR mRNA degradation in murine embryonic fibroblasts treated with β1‐AR agonists and antagonists with overexpression and/or siRNA‐mediated silencing of HuR and AUF1. These findings provide important insight into the regulation of the β1‐AR in heart failure and may suggest new therapeutic targets for the restoration of healthy adrenergic function.
CaMKIIδ splice variants exert differential effects on heart failure development and myocardial ischemia/reperfusion injury (652.14)
Authors: Gray Charles , Xiang Sunny , Mishra Shikha , Heller Brown Joan
Published: June 2021

Mice that lack CaMKIIδ (KO) are significantly protected from the development of myocardial damage following ischemia reperfusion (I/R). Cardioprotection in KO animals was associated with the loss of NF‐κB‐mediated inflammatory responses. CaMKIIδ is alternatively spliced in the heart to generate the CaMKIIδB and δC subtypes. In vitro data also suggest that δC is deleterious while δB may be cardioprotective. Furthermore, mice expressing δC rapidly and spontaneously progress to heart failure and death while those expressing δB have a milder phenotype. We postulated that CaMKIIδB and δC play opposing roles in response to I/R injury. Mice expressing CaMKIIδB and δC were subjected to global ischemia in the Langendorff mode for 25 minutes followed by reperfusion for various times. CaMKII activation and substrate phosphorylation were assessed by western blot. Subcellular fractionation was performed to determine compartment‐specific CaMKII activation. Infarct size was used as a measure of myocardial injury. I/R led to activation of CaMKIIδC but not δB in the cytosolic compartment. IκB‐Kinase (IKK) phosphorylation was increased in mice expressing CaMKIIδC but not in those expressing δB. Mice expressing δC showed large infarcts compared to WT, while mice expressing CaMKIIδB show greatly diminished infarct formation following I/R. The observation that pharmacological inhibition of IKK mitigated the deleterious effects of δC overexpression suggests that selective activation of CaMKIIδC in the cytosol and subsequent activation of inflammatory signaling contributes to I/R injury.

Grant Funding Source: Supported by NIH Grant HL080101
Development of pharmacological and genetic tools to explore the potential of inward rectifier potassium channels as novel insecticide targets (654.9)
Authors: Raphemot Rene , Estevez‐Lao Tania , Rouhier Matthew , Hillyer Julián , Hopkins Corey , Piermarini Peter , Denton Jerod
Published: June 2021

Inward rectifying potassium (Kir) channels play important physiological roles in humans and are expressed in various insect species such as mosquitoes. However, their potential as insecticidal targets remain largely unknown. Current chemical vector control strategies are becoming less effective due to the emergence of insecticide resistance. We have shown recently that impairment of Aedes aegypti Kir1 channels with a small molecule inhibitor elicits “renal failure” and death in culicine mosquitoes, and thus could represent a novel insecticidal target. We are employing medicinal chemistry to improve the potency and physicochemical properties of new Kir1 channel inhibitors identified via high‐throughput screening and assessing their effects on mosquito mortality. Quantitative RT‐PCR analyses revealed that Anopheles gambiae Kir1 is expressed in various tissues and is highest in the ovaries. To elucidate a putative role of Kir1 channels in anopheline mosquito reproduction, we developed a RNA interference strategy to knockdown Kir1 expression and assess its effects on mosquito fecundity. The use of these novel molecular and pharmacological tools will be useful to uncover novel physiological functions and the insecticidal potential of Kir channels in vectors of disease‐causing pathogens. Funded by a grant from the Foundation for the NIH, VCTR program.
Electrically monitoring effects of cisplatin on transendothelial invasion of human ovarian cancer cells (655.11)
Authors: Lo Jun‐Chih , Lo Chun‐Min
Published: June 2021

Cisplatin is a platinum based chemotherapy drug used to treat various types of cancers, including sarcomas, some carcinomas, lymphomas and germ cell tumors. In this study human ovarian cancer cell lines, OVCA429 and ALST, were challenged with different concentrations of cisplatin (1E‐6M, 1E‐5M, 5E‐5M and 1E‐4M) and the effects on cancer cell morphology and micromotion were monitored and analyzed by electric cell‐substrate impedance sensing (ECIS) and microscopy. Moreover, the transendothelial invasion of the ovarian cancer cells to human umbilical vein endothelial cells (HUVECs) was examined. Our measurements showed that 1E‐4M of cisplatin caused a decrease of junctional resistance between cells and the reduction in micromotion. In addition, 1E‐6M and 1E‐5M of cisplatin were not toxic to the integrity of HUVEC layers but effectively inhibited transendothelial invasion of ovarian cancer cells. Thus, we report a strategic approach for using ECIS to analyze potential drugs for their abilities to suppress metastasis, particularly at the level of transendothelial invasion.

Grant Funding Source: Supported by the National Science Council in Taiwan
Unmasking the the mechanism of action of some medicinal plants used in diabetes mellitus (657.20)
Authors: Obih Patience , Summers Brittinni , Ezebuenyi Michael , Jimoh Akeem , Ambush Evelyn
Published: June 2021

Diabetes mellitus is a chronic and metabolic disease affecting many parts of the world. It affects over 347 million people globally and about 25.8% of the American population. The drugs in current use for the management of diabetes mellitus are not free from side effects. As a result of limitations in many conventional antidiabetic agents, researchers are now investigating traditional remedies for better alternatives for diabetic treatment. Blueberry and dandelion are used as food and they have been reported by some workers to have therapeutic effect on diabetes mellitus. Some antidiabetic agents e.g. Acarbose reduce conversion of starch and disaccharides to monosaccharides, thereby reducing postprandial hyperglycemia. They do this by inhibiting alpha glucosidases, but they have their side effects. The objective of this study is to examine the alpha glucosidase inhibitory activity of some medicinal plants like blueberry and dandelion. The inhibitory effect of different concentrations of aqueous extracts of blueberry and dandelion separately on alpha glucosidase was studied in vitro. The inhibition study was carried out spectrophotometrically in a 96‐well plate and the absorbance was measured using a Thermo Scientific Multiscan spectrophotometer. Our results indicate that these extracts inhibited alpha glucosidase (EC50 of 32 mg/ml for blueberry and 45 mg/ml for dandelion). In conclusion, blueberry and dandelion inhibited alpha glusidase in our system. Acarbose and miglitol with similar mechanism of action are already marketed for the treatment of diabetes. Observations made in this project will go a long way to furnish more information needed for introduction of better alternatives in managing diabetes. However more studies are needed.
The soluble epoxide hydrolase as a novel target for chronic neuropathic pain (659.15)
Authors: Wagner Karen , Inceoglu Bora , Hammock Bruce D.
Published: June 2021

Neuropathic pain remains an unmet clinical need. In addition to nerve injury cases of neuropathy diabetes is increasing worldwide and diabetic neuropathy, the most frequent secondary condition to diabetes, will increase with it. Recent investigations reveal lipids as signaling mediators and their functional significance in nociception is receiving attention. Notably, there is evidence of antihyperalgesia mediated by epoxy fatty acids (EpFAs), metabolites generated by cytochrome P450 epoxygenase in an arm of the arachidonic acid cascade. These endogenous lipid mediators are subsequently degraded by the soluble epoxide hydrolase (sEH) enzyme to their mostly inactive diols. Metabolomic studies show that administration of sEH inhibitors (sEHI) stabilize EpFA to diol ratios in vivo and lead to decreased inflammation and pain related behavior in inflammatory pain models. Unexpectedly, the sEHI are also active against a model of type I diabetic neuropathy where the role of inflammation is minimal. The experiments here in rat and mouse demonstrate that potent sEHI mediate antihyperalgesia in both the sciatic nerve ligation (SNL) neuropathic pain and diabetic neuropathic pain (DNP) models. While mechanical withdrawal thresholds were used to assess both the SNL and DNP modeled pain, there are limitations to using acute stimulus reflex responses to assess chronic neuropathy. Therefore, the conditioned place preference (CPP) assay was used to evaluate the tonic nature of chronic pain and to address these limitations. Here, the CPP assay alongside withdrawal thresholds demonstrates the dose dependent antihyperalgesic efficacy of sEHI in the murine DNP model. Importantly, the CPP effect was absent both in control mice and in sEH knockout mice indicating the sEHI is not positively reinforcing or rewarding. Overall, these experiments indicate that inhibiting the sEH attenuates neuropathic pain and offers an alternative to current therapies to meet this clinical need.

Grant Funding Source: Supported by R01 ES002710, P42 ES004699, R21AR062866
Role of cAMP signaling in mucin production in response to IL‐13 in human bronchial epithelial cells (660.12)
Authors: Al‐Sawalha Nour , Senny Christina , Hernandez Adrian , Valdez Daniel , Pokkunuri Indira , Thanawala Vaidehi , Bond Richard , Knoll Brian
Published: June 2021

Asthma, a chronic airway inflammatory disease, is associated with mucus hypersecretion by airway epithelium. Accumulated mucus leads to airway obstruction and morbidity. We have shown that β2AR signaling has pro‐inflammatory effects in cultured normal human bronchial epithelial cells (NHBECs) in response to IL‐13(1). Our objective here was to elucidate the role of cAMP signaling in mucin production.

NHBECs growing at air‐liquid interface (ALI) were subjected to various treatments, and levels of MUC5AC transcripts, intracellular mucin, cAMP levels, pCREB and pSTAT6 were measured on day 14 ALI.

Treatment with forskolin+IBMX (to raise global intracellular cAMP levels) increased MUC5AC transcripts and intracellular mucin in the presence or absence of epinephrine in response to IL‐13 pCREB and pSTA6 were also elevated only in the presence of epinephrine, IL‐13 and forskolin+IBMX. A compartmentalized increase in cAMP caused by treatment with roflumilast, a PDE4 inhibitor, did not affect MUC5AC expression but reduced intracellular mucin only in the presence of epinephrine+IL‐13. Roflumilast did not affect the levels of cAMP, pCREB or pSTAT6.

Our data provide evidence for the different influences of global and compartmentalized increases in cAMP on mucin production in human airway epithelium in response to IL‐13.

1.Pokkunuri et al, BPS, 2011 Winter Meeting

Grant Funding Source: Supported by NIH 1RO1A179236 awarded to RAB and BJK
Modulation by tetrodotoxin of action potential discharge in guinea pig vagal tracheal nodose afferent A‐delta cough receptors (660.15)
Authors: Herbstsomer Adam , Meeker Sonya , Undem Bradley
Published: June 2021

We used the isolated vagally innervated guinea pig trachea/bronchus preparation to characterize the inhibitory effect of tetrodotoxin (TTX) on action potential discharge evoked by electrical (concentric stimulating electrode), and chemical (citric acid) activation of capsaicin‐insensitive nodose A‐delta fibers. We previously described these nerves and bona fide “cough receptors” (Canning et al. J. Physiol, 2004). We have also previously noted at the mRNA, protein, and electrophysiological level that the cell bodies of these nerves express TTX‐sensitive (mainly NaV 1.7) and TTX‐insensitive (both NaV 1.8 and NaV 1.9) channels (J. Physiol. 2008). Here we evaluated 18 nodose fibers with mean CVs of 5.1 ± 0.5 m/s. The voltage threshold of the terminals was 13 ± 1.5 V. The peak following Hz (>95% success for 5s) averaged 30 ± 2 Hz. All fibers responsed to citric acid with an average peak AP discharge of 35 ±6 Hz. Over 95% (17/18) of the fibers were completely silenced with 1 µM TTX. At 0.1 µM, TTX increased the voltage threshold to 24 ± 4 V, reduced the acid response to 10 ± 2 Hz, but had no effect on peak following Hz (studied at 2x V threshold). Jugular neurons project capsaicin‐sensitive A‐delta fibers with CVs the same as the nodose fibers (CV = 6.2 ± 1.4 m/s) and C‐fibers (CV = 0.6 ±0.1 m/s) to the trachea/bronchi. In contrast to nodose A‐delta fibers, only about 50% of jugular A‐delta (5/11) or C‐fibers (5/9) fibers were silenced with 1 µM TTX. These data indicate that although the TTX‐resistant NaVs (1.8 and 1.9) are expressed in nodose tracheal A‐delta cough receptors , they cannot alone support action potential discharge in healthy animals; this is in contrast to the jugular A‐delta and C‐fibers in the trachea/bronchi where TTX‐resistant channels may play a more important role.

Grant Funding Source: NIH, Bethesda Maryland, USA
Alveolar type II cell‐localized p120‐catenin is an essential regulator of lung barrier function and innate immunity (660.8)
Authors: Chignalia Andreia , Vogel Stephen , Reynolds Albert , Malik Asrar , Liu Yuru
Published: June 2021

Maintenance of alveolar epithelial barrier (AEB) integrity is crucial for normal lung gas exchange function, fluid balance, and regulation of immune response. The mechanisms whereby AEB is kept are not clear but adherens junctions (AJ) are central. P120‐catenin (p120) is an important component of AJ that stabilizes E‐cadherin. The alveolar epithelium is composed of two types of cells: flat type I cells comprising ~95% of the gas‐exchange surface, and cuboidal type II cells (ATII), which secrete surfactant and play a role in modulating lung immunity. Here we addressed if p120 contributes to ATII regulation of AEB and immunity by developing an inducible ATII‐specific p120 knockout mouse (P120EKO). P120EKO lungs displayed persistently leaky alveolar barrier and chronic lung inflammation. The inflammatory phenotype included infiltration of inflammatory cells, increased MPO and NFκB activity, and enhanced ICAM1and MIP2 expression. p120EKO ATII displayed E‐cadherin instability and decreased transepithelial resistance. p120EKO lungs showed barrier leakiness as shown by increased 125I‐BSA permeability in vivo. P120EKO lungs had increased surfactant proteins (SpB, SpC and SpD) and TLR4 expression. These findings indicate that while ATII constitute only 5% of alveolar surface area, ATII‐localized p120 plays a critical role in maintenance of alveolar epithelial barrier and homeostasis and modulate lung immune response.

Grant Funding Source: Supported by CAPES fellowship (AZC), NIH grants HL105947‐01 (YL), HL07829‐16 (AM), HL090152 (AM)
The effect of aging on microvascular density and angiogenesis (665.4)
Authors: Sweat Richard , Chedister Lee , Sloas David , Stewart Scott , Murfee Walter
Published: June 2021

Age related pathologies are commonly associated with vessel loss or excess vessel growth. An important question is whether or not angiogenesis, defined as the growth of new vessels from existing ones, is altered during aging. Angiogenesis in aged populations is commonly thought to be impaired. However, much of this supporting evidence is based on local changes in angio‐regulator molecules. The objective of this study was to quantitatively compare microvascular network remodeling metrics during angiogenesis in adult versus aged rat strains. Mesenteric tissues from adult (9 months) and aged (24 months) male Fischer 344 rats were harvested according to 4 experimental groups: 1) Adult Unstimulated (n=4 rats, 16 tissues), 2) Aged Unstimulated (n=4 rats, 16 tissues), 3) Adult Stimulated (n=4 rats, 16 tissues), 4) Aged Stimulated (n=4 rats, 16 tissues). For stimulated groups, tissues were harvested 3 days post compound 48/80‐induced mast cell degranulation stimulation. Unstimulated aged microvascular networks displayed larger mean vascular area per tissue area compared to the unstimulated adult networks. Following angiogenic stimulation, both adult and aged networks displayed similar increases in vascularized area and vessel length density compared to their respective unstimulated control groups, indicating that both adult and aged networks are capable of undergoing angiogenesis. However, capillary sprouting was significantly lower in stimulated aged networks compared with stimulated adult networks, supporting the hypothesis of impaired angiogenesis during aging. Interestingly, our results provide metric specific evidence for both comparable and impaired angiogenesis during aging and offer a possible explanation for conflicting results from the literature.

Grant Funding Source: Supported by Tulane Center for Aging and NIH 5‐P20GM103629‐02
A novel low pHo‐induced Cl‐ conductance in nodose ganglion neurons is neuroprotective during simulated ischemia: implication for vagal afferent activity in heart failure (667.9)
Authors: Wang Runping , Lu Yongjun , Cicha Michael , Chapleau Mark , Benson Christopher , Abboud Francois
Published: June 2021

We found that vagal afferent neurons isolated from nodose ganglia exhibit a large and prolonged low pHo‐conditioned Cl‐ current (25±5 pA/pF, n=60), when exposed to 3 brief pulse of low pHo (6.0) solution delivered at 20sec intervals. The current was similar to the one carried by the “Swell‐Activated Cl‐ Channel” (SAC) during hypoosmolarity. The activation pathways of this Cl‐ conductance are through NADPH oxidase (NOX)‐dependent production of ROS (dihydroethidine fluorescence). Since a similar Cl‐ conductance was found to be protective during ischemic preconditioning in cardiomyocytes (Draz et al 1999) we tested the hypothesis that the activation of this current may also be protective in cultured neurons exposed to “simulated ischemia” (low glucose, hypoxia and pH5.0). We measured the LDH release as an indication of cell injury. Without low pHo preconditioning the supernatant LDH concentration averaged 82±4 μU/ml and 441±62 μU/ml after exposure to 1 and 3 hours of simulated ischemia. In contrast, supernatant LDH was significantly reduced to 58±10 μU/ml and 227±25 μU/ml at 1 and 3 hours when neurons were preconditioned by activating the Cl‐ current before simulated ischemia. The protective effect of preconditioning was partially reversed by SAC channel inhibitor tamoxifen (10μM) (393±99 μU/ml at 3hours).

Conclusions and Clinical Implications: A low pHo‐induced activation of a Cl‐ conductance in nodose neurons is NOX mediated and provides a putative autocrine protective mechanism for preservation of cardiac vagal afferents during ischemia or heart failure.

Grant Funding Source: Supported by NIH HL 14388
Evidence for altered DNA methylation as a major regulator of gene expression in calcific aortic valve disease (671.15)
Authors: Nwachukwu Nneka , Hagler Michael , Kafa Rami , Roos Carolyn , Miller Jordan
Published: June 2021

Aging is a major risk factor for calcific aortic valve disease (CAVD). While recent work suggests that DNA methylation patterns are dramatically altered with aging, changes in DNA methylation patterns in age‐related cardiovascular diseases remain largely uncharacterized. Thus, we tested the working hypothesis that de novo DNA methyltransferase levels (DNMT3b in particular) are significantly increased in stenotic human aortic valves and would serve to repress expression of protective genes. We used human aortic valves that were either normal (NORM) or stenotic (STEN). qRT‐PCR and Western blotting were used to examine analyze changes in DNMT3b levels and markers of osteogenesis. DNA methylation was examined using ELISA‐based methods for total methylation and Illumina 450k Beadchip assays for site‐specific methylation between NORM and STEN valves. The pro‐osteogenic marker expression, Sp7, was increased by 4±1 fold in STEN valves compared to NORM tissue. While mRNA levels of DNMT3b only tended to be increased in STEN valves, protein levels of DNMT3b were dramatically increased in STEN valves compared to NORM valves (13.9±3‐fold increase). Increases in DNMT3b levels were associated with an increase in global DNA methylation, and 450k beadchip analysis identified more than 6,000 differentially‐methylated sites between NORM and STEN tissue. Collectively, these data identify DNA methylation as a potentially novel and major regulator of gene expression in end‐stage calcific aortic valve stenosis.

NHLBI

Grant Funding Source: NHLBI
Effects of stenosis on hemodynamics in carotid artery bifurcation using 2D MR‐imaging computational fluid dynamics: simulated versus real cases (671.4)
Authors: Chatpun Surapong , Prakobkarn Arpapan , Ina Natee , Saeheng Sakchai
Published: June 2021

Carotid artery stenosis is one of major risks of stroke. Magnetic resonance (MR) imaging combined with computational fluid dynamics becomes a powerful tool to investigate hemodynamic changes in circulatory system for specific patients. In this study, hemodynamics obtained from a real stenotic model of carotid artery from two patients was compared with a simulated stenotic model from two healthy subjects. MR angiographic image of carotid artery was reconstructed as an input geometry for two‐dimensional computational fluid dynamics. Inlet velocity used in the calculation was obtained from Doppler ultrasound measurement. This calculation was based on the simple assumptions i.e. incompressible and Newtonian fluid, rigid vascular wall and non‐slip condition at wall. The computational results showed that stenosis in common carotid artery increased blood flow velocity in internal carotid artery which was higher than that in external carotid artery. Increased severity of stenosis also enhanced blood flow velocity but lowered wall shear stress on the inner wall of internal carotid artery. Multiple stenotic points decreased velocity of internal carotid artery compared with single point stenosis. This study demonstrated that the simulated stenotic model of carotid artery from healthy subjects has conditionally used to represent the effects of stenosis found in the real stenotic model.

Grant Funding Source: Supported by Facualty of Medicine, Prince of Songkla University and NSTDA
Evaluation of mesenchymal stem cell contribution to adipose health in the context of high fat diet and exercise (673.2)
Authors: Pincu Yair , Huntsman Heather , Zou Kai , De Lisio Michael , Mahmassani Ziad , Boppart Marni
Published: June 2021

Objective: To investigate the role of mesenchymal stem cells (MSCs) in adipose tissue (AT) inflammation and fibrosis during high fat diet and exercise training. Methods: C57BL/6 mice were fed high fat (HF, 60%) or low fat (LF, 10%) diets for 8 weeks. Half of the mice were then exercised for 16 weeks. Sca1+CD45‐ MSCs were extracted from epididymal fat, isolated by FACS and analyzed by Q‐PCR. AT inflammation and fibrosis were evaluated using Q‐PCR and histology. Results: HF diet (HFD) increased body weight and exercise attenuated weight gain. AT F4/80, TNFα, and IL‐10 gene expression increased with HF (p<0.05). Exercise did not attenuate this increase. Genes associated with fibrosis (TGFβ1 and Col1α) increased with HFD, and Col1α expression was attenuated with exercise (p<0.05). HFD decreased the percentage of MSCs in AT, but the total number was higher compared to LF fed mice (p<0.05). Exercise did not impact MSC content in AT. MSC gene expression was altered by HFD, but not by exercise. Specifically, Col1α, MMP2, Timp1,VEGFa and IGF‐1 increased while MMP9 decreased (P<0.05). MSC inflammatory gene expression was not altered with HFD or exercise. Conclusion: MSCs do not contribute to AT inflammation, but do express genes necessary for extracellular matrix remodeling and angiogenesis in obese mice. Overall, these data suggest that MSCs provide a mechanism for AT expansion when challenged by HFD.
Loss of cerebral blood flow autoregulation and assessment of the critical cerebral perfusion pressure at high ICP (674.14)
Authors: Nemoto Edwin , Bragin Denis , Statom Gloria , Bragina Olga
Published: June 2021

Objective: To study the effect of increased intracranial pressure (ICP) on the critical cerebral perfusion pressure (CPP) and its relationship to the loss of cerebral blood flow (CBF) autoregulation.

Background: Earlier studies showed that the critical cerebral perfusion pressure (CPP) fell from 60 to 30 mmHg as determined by decreasing arterial pressure as opposed to increasing intracranial pressure (ICP), respectively. We hypothesized that the decrease in the critical CPP at high ICP was due to microvascular shunting resulting in a pathological higher CBF at a lower CPP.

Methods: Using two‐photon laser scanning microscopy (2PLSM) we measured microvascular (3‐25 m diam) flow velocity in rat cerebral cortex by increasing ICP or decreasing arterial blood pressure. In separate studies we also increased ICP progressively while monitoring the capillary (CAP) to microvascular shunt (MVS) ratio to study the effect of increasing ICP on the development of microvascular shunts.

Results: The reduction in critical CPP with increasing ICP was due to increased microvascular shunting resulting in a sustained CBF at a lower critical CPP, which therefore decreased the critical CPP from 60 to 30 mmHg and associated with increased microvascular shunting. A progressive increase in ICP resulted in a parallel increase in the ratio of MVS/CAP ratio.

Conclusion: The decrease in critical CPP obtained when ICP is increased as opposed to decreasing arterial pressure is due to the opening of microvascular shunts thereby creating low resistance microvascular conduits through the brain. The progressive increase in MVS with increasing ICP leads to the loss of CBF autoregulation, which differs from that observed at high CO2.

Grant Funding Source: NIH RO3 NS051639;AHA 2BGIA11730011
Ultrasound‐mediated nanoparticle delivery across the blood‐brain barrier (676.17)
Authors: Timbie Kelsie , Nance Elizabeth , Zhang Clark , Miller Wilson , Song Ji , Hanes Justin , Price Richard
Published: June 2021

Introduction: The intact blood‐brain barrier (BBB) presents a major obstacle for drug delivery to the brain, as it prevents the vast majority of therapeutics from entering brain tissue. Noninvasive focused ultrasound (FUS) in conjunction with microbubbles (MB) has been shown to cause safe, reversible, localized disruption of the BBB. This work examines the ability of MR guided FUS to deliver specialized brain‐penetrating nanoparticles (NP) across the BBB. NPs were 60 nm in diameter and covered with a dense PEG coating to permit diffusion through brain tissue.

Materials and Methods: Anesthetized rats were coupled to the 1.14 MHz FUS system and given an IV co‐injection of fluorescent NPs and MBs immediately before sonication. Peak negative pressure was either 0.4 MPa or 0.6 MPa. Immediately following sonication, T1‐weighted contrast enhanced MRI images were captured to verify BBB disruption. Animals were placed on a heating pad at 37°C for one hour to allow increased NP uptake. Mounted brain sections were stained with BS‐I lectin to reveal endothelial cells (ECs) and imaged with confocal microscopy.

Results and Discussion: Treatment with 0.6 MPa tripled NP coverage within the brain and produced larger NP “clouds” extending into the brain parenchyma, in addition to significantly increasing the occurrence of large NP clouds (>100 μm2) Furthermore, treatment with higher US pressure increased NP delivery to brain parenchyma rather than to the endothelial cells (ECs) lining the vasculature. Control regions showed no NP delivery.

Conclusions: Our results indicate that varying US pressure can divert NP delivery between the endothelium and brain parenchyma.

Grant Funding Source: Supported by NIH R01 CA164789 and The Focused Ultrasound Surgery Foundation