bluuebunny/crossref_metadata_2025 huggingface dataset:
Familiarity, perceptions, and knowledge of commonly consumed ethnic spices among registered dietitians and dietetic students
Authors: Fernandes Nicolle V , Chezem Jo Carol , Kandiah Jayanthi
Published: June 2021

The health benefits of Complementary and Alternative Medicine (CAM) are gaining popularity. The practice of CAM includes herbs and spices. Registered dietitians (RDs) play a unique role in influencing their use. This study evaluated familiarity, perceptions, and knowledge of ethnic spices among RDs and Dietetic Students(DS). At a Midwestern dietetics conference, 144 participants (83 RDs and 61 DS) completed a 13‐item survey on black pepper, chili pepper, cilantro/coriander, cinnamon, clove, curry leaves, fenugreek, garlic, ginger, and turmeric. Among respondents, 89% were familiar with seven (P<0.05); fewer than 78%, 57% and 30% were familiar with turmeric, curry leaves and fenugreek, respectively. Cinnamon, garlic and ginger were perceived as most effective in health promotion and prevention/treatment of diseases (P<0.05). Most frequent associations made between spices and diseases were cinnamon with diabetes (71%), garlic with cardiovascular disease (50%) and ginger with gastrointestinal disorders (49%). Barriers to the use of spices included insufficient education (80%) and lack of scientific evidence (62.5%). All participants indicated a need for spice education. This may be through continuing education and academic preparation for RDs and DS, respectively.
Supplementation of reinforced galactooligosaccharides modulates intestinal l microbiota
Authors: Hong Ki‐Bae , Kim Kwang‐Min , Park Yooheon , Suh Hyung Joo
Published: June 2021

A dietary prebiotic is an important fermented ingredient in host health that is essential for positive microbiota composition, intestinal functions and immune system. Based on many studies so for, prebiotic effects associated with a selective stimulation of growth of one or a limited number of microorganisms and several prebiotics are now widely used commercially in food. In the present study, our aim was to characterize high concentration of galactooligosaccharides(GOS) and relationship with bifidobacteria. Galactooligosaccharides(GOS) are one of prebiotic groups that beneficially affect intestinal microbiota, barrier functions, and other health benefiting effects in the human and animal studies. However a line of recent studies did not suggest that effects of high content GOS between the fecal microbiota and intestinal regulation. Our present study utilizing whole blood measurements, fecal bacteria populations, and molecular markers. These results support a direct relationship between concentration of GOS and beneficial bacteria activity. We propose the hypothesis that prebiotic properties of high content GOS could use potential food application.
Protective effect of Cirsium setidens extract against the alcohol‐induced liver injury in rats
Authors: Kim Eun‐hye , Chung Jayong
Published: June 2021

Cirsium setidens is a medicinal herb that is rich in flavonoids. The aim of this study was to determine the effect of Cirsium setidens ethanol extract (CSE) against the alcohol‐induced liver injury in rats. Sprague‐Dawley rats were fed a Lieber‐DeCarli liquid diet with either alcohol alone, alcohol +100 mg/kgBW CSE, or alcohol + 500mg/kgBW CSE for 2 months. Control rats were fed the same amount of isocaloric liquid diet without alcohol. Chronic alcohol feeding resulted in the development of hepatic steatosis, and significantly increased the levels of ALT and AST in the serum, as compared with controls. The alcohol‐induced steatosis was associated with the up‐regulation of fatty acid synthase (FASN) gene expression. Further, alcohol feeding greatly stimulated the hepatic NF‐κB signaling pathway, a master regulator of inflammation. In contrast, CSE supplementation in alcohol‐fed rats ameliorated hepatic steatosis, and restored the levels of serum ALT and AST to control levels. Moreover, the CSE supplementation significantly blocked the alcohol‐induced hepatic FASN gene expression in a dose‐dependent manner. In addition, the CSE supplementation markedly suppressed the activation of NF‐κB in the liver tissues of alcohol‐fed rats. In summary, our study results suggest that CSE protects against alcohol‐induced liver injury, via the suppression of fatty acid synthesis and inflammation.
Effects of a leucine and pyridoxine‐containing nutraceutical on body weight and composition in obese adults
Authors: Zemel Michael B. , Bruckbauer Antje
Published: June 2021

We recently demonstrated leucine to modulate energy partitioning between adipose tissue and muscle. Further, leucine exhibits a synergy with B6, resulting in reduced adipocyte lipid storage coupled with increased muscle fat oxidation. A nutraceutical (NuShape) containing 2.25 g leucine and 30 mg B6 increased fat oxidation by >;30 g/d in a 28‐day RCT. To evaluate the efficacy of this combination in modulating body weight and composition, we conducted two 24‐week placebo‐controlled RCTs, one with weight maintenance (n=20) and one hypocaloric (−500 kcal/d; n=24). The supplement resulted in fat loss in the maintenance study (−1.12±0.36 and −1.82±0.70 kg at 12 and 24 weeks, p<0.01 vs placebo) while no change was found in the placebo‐group. In the hypocaloric study, the supplement group lost up to twice as much weight (6.18±1.02 vs. 3.40±0.81 kg at 12 weeks and 8.15±1.33 vs. 5.25±1.13 kg at 24 weeks, p<0.01) and fat (4.96±0.61 vs. 2.31±0.53 kg at 12 weeks and 7.00±0.95 vs. 4.22±0.74 kg at 24 weeks, p<0.01) than placebo. Thus, this nutraceutical combination results in significant fat loss in the absence of caloric restriction and markedly enhances weight and fat loss by 50–80% over a 24‐week period.
The Impact of Nutrition Education on Food Security Status and Food‐Related Behaviors
Authors: Farrell Jamie Allison , Cordeiro Lorraine , Qian Jing , Anliker Jean , Peterman Jerusha
Published: June 2021

Low‐income households are at risk of food insecurity and altered food related behaviors. This study examined the relationship between food security status and specific food purchasing practices. Participants in the Massachusetts Expanded Food and Nutrition Education Program (EFNEP) completed a pre‐curriculum survey. Household food security was assessed by the USDA 6‐item food security module. We asked additional questions about how often participants could afford healthy foods, fruits and vegetables, and the same foods throughout the month. In the pre‐EFNEP survey, 41% (n=58) were food insecure and 66% (n=38) had marginal/low/very low food security. Overall, 22% (n=13) that could always or often afford healthy foods, 26% (n=15) that could always or often afford fruit and vegetables, and 24% (n=14) that could always or often afford the same foods all month experienced low/very low/marginal food security. Those with marginal/low/very low food security were less likely than fully food secure to report they could always afford healthy foods, fruits and vegetables, and the same foods all month. Food purchasing behaviors can be affected by the strain of limited resources. Nutrition education can provide skills and knowledge to low‐income, food insecure populations so they may better manage resources to afford foods they need throughout the month.

UMass Amherst Office of Faculty Development
Women who participate in a structured weight loss program with resistance‐exercise experience more favorable changes in body composition when compared to other popular weight loss programs
Authors: Lockard B , Baetge C , Levers K , Galvan E , Jagim A , Simbo S , Byrd M , Jung YP , Oliver J , Koozehchian M , Dalton R , Khanna D , Sanchez B , Kresta J , Horrell K , Leopold T , Cho M , Springer S , Rivera A , Cerda C , Rasmussen C , Kreider R
Published: June 2021

127 sedentary women (46±12 yr, 45.5±5% body fat, 35.1±5 kg/m2) were randomized to participate in a control group (C) or the Curves Complete® program with online support (CC), Weight Watchers® Points Plus (WW), Jenny Craig® (JC), or Nutrisystem® Advance Select™ (NS) weight loss programs for 12‐wks. Body mass, DEXA body composition, and resting energy expenditure (REE) measurements were obtained at 0, 4, 8, & 12 wks and analyzed by MANOVA. Data are presented as changes from baseline after 12‐wks. Participants in the diet groups lost similar amounts of body mass (C 0.1±3.1; CC −6.1±3.2; WW − 4.3±3.5; JC −5.3±3.8; NS −5.1±4.5 kg, p=0.001). However, participants in the CC group experienced significantly greater loss in fat mass (C −0.0±2.0; CC −5.2±2.8; WW −2.2±2.6; JC −3.5±3.3; NS −2.3±2.5 kg, p=0.001), less loss in FFM (C 0.1±2.3; CC − 0.7±2.5; WW −1.8±2.3; JC −1.8±2.1; NS −2.4±2.2 kg, p=0.002), and greater reductions in percent body fat (C −0.1±1.7; CC − 3.3±2.3; WW −0.6±2.5; JC −1.4±2.4; NS −0.2±1.7%, p=0.001). REE tended to differ among groups (C 39±190; CC 0±154; WW − 108±159; JC −27±206; NS −43±206 kcal/d, p=0.07). Results indicate that participation in the CC program promotes more favorable changes in body composition than other popular weight loss programs.
Pediatric patients with Prader‐Willi syndrome have lower muscle mass than those with exogenous obesity
Authors: Buraso Chutdanaiphakorn , Wattanasirichaigoon Duangrurdee , Sae‐Chew Pattarana , Phoonlapdacha Phanphen , Dumrongwongsiri Oraporn , Sangcakul Areeporn , Suthutvoravut Umaporn , Chongviriyaphan Nalinee
Published: June 2021

Patients with Prader‐Willi syndrome (PWS) have morbid obesity and short stature. We conducted this cross‐sectional study to determine body composition in pediatric patients with PWS.

We enrolled 3 groups of children aged 2–15 years: 12 diagnosed with PWS, 18 with exogenous obesity, and 18 healthy with normal weight for height as controls. Body composition was determined using bioelectrical impedance analysis (BIA).

We found that the PWS group had significantly higher ratio of fat mass to fat‐free mass than the exogenous obesity group and normal controls (1.1, 0.6, and 0.2, respectively; p = 0.001 and less than 0.001, respectively). The median percentage of muscle mass in the PWS group was significantly lower than that in the exogenous group (47 vs. 58%; p = 0.003) and that in the normal control group (47 vs. 80%; p < 0.001).

In conclusion, pediatric patients with PWS had higher fat mass and lower muscle mass than those with exogenous obesity. The intervention for this group of patients should include not only controlling weight but also increasing muscle mass.

This study was funded by the Nutrition Association of Thailand (Nestle Nutrition Institute Research Award).
Maternal/household characteristics and child stunting in Ethiopia: A secondary analysis of the 2011 Ethiopian Demographic and Health Survey
Authors: Girma Meron , Stoecker Barbara J
Published: June 2021

Characteristics related to child (<60 months, n=9,892) stunting and underweight were examined using data from the nationally representative 2011 Ethiopian Demographic and Health Survey. Stunting (height‐for‐age Z < −2) and underweight (weight‐for‐age Z < −2) were categorized using World Health Organization standards. Both stunting and underweight were significantly associated with maternal education, BMI, residence and wealth. Children of underweight mothers (BMI < 18.5) were significantly more likely [adjusted odds ratio (AOR)=1.16] to be stunted than children of mothers with normal BMI. Furthermore, mothers with no formal education [AOR=1.67] and mothers with primary education [AOR=1.52] were significantly more likely to have a stunted child than mothers with secondary or tertiary education. Rural children were more likely than urban to be stunted [AOR=1.34], and children from the lowest wealth category were more likely to be stunted than those from the highest wealth category [AOR=1.57]. Likewise for underweight, mothers with no education were more likely to have an underweight child [AOR= 2.05] compared to mothers who had more than primary education. Children from the lowest wealth category were nearly twice as likely [AOR=2.04] to be underweight compared to children from the highest category. Maternal and household characteristics remain important predictors of child nutritional status in Ethiopia. (Supported by Oklahoma State University)
Dietary patterns, instant noodles intake, and cardiometabolic risk factors
Authors: Shin Hyun Joon , Cho Eunyoung , Lee Hae‐Jung , Fung Teresa , Rimm Eric , Rosner Bernard , Manson JoAnn E , Hu Frank
Published: June 2021

To find the relationship between dietary patterns, instant noodles and cardiometabolic risk, we analyzed 10,711 adults (females 54.5%) between 19–64 years of age in the Korean National Health and Nutrition Examination cross‐sectional Survey (KNHANES) IV 2007–2009. A “meat and fast food pattern”(MP) rich in intakes of meat, soda, fried food, and fast food including instant noodles and a “traditional pattern”(TP) rich in intakes of fruit, vegetables, and fish were identified with principal component analysis. The highest MP quintile was associated with higher odds for abdominal obesity(OR:1.41, 95% CI 1.05–1.90), LDL≥130mg/dL(OR:1.57, 95% CI 1.26–1.95), lower odds for low HDL(OR:0.64, 95% CI 0.52–0.78), and TG≥150mg/dL(OR:0.73, 95% CI 0.57–0.93), but was not associated with metabolic syndrome. The highest TP quintile was associated with lower odds for elevated blood pressure(OR:0.73, 95% CI 0.59–0.90), marginally lower trends of abdominal obesity(p trend 0.055) and in males, was associated with lower trend of metabolic syndrome (p trend 0.049). Two or more servings/week consumption of instant noodles was associated with a higher odds for metabolic syndrome (OR:1.68, 95% CI 1.10–2.55)in females, but not in males(p for interaction 0.04). In conclusion, instant noodles were associated with metabolic syndrome independent of major dietary patterns especially in females.
Anti‐inflammatory and anti‐hyperalgesia activities of KMUP‐1 in a rat chronic constriction injury model
Authors: Wu Bin‐Nan , Lin Ting‐Chun , Chia Soo‐Lin
Published: June 2021

Neuropathic pain is characterized by spontaneous pain, hyperalgesia and allodynia. Pharmacological interventions have had limited success so far. In this study, we investigate whether KMUP‐1 could improve pain hypersensitivity and reduce inflammatory mediators, and also explore the possible mechanisms in rats’ sciatic nerve with chronic constriction injury (CCI) to induce neuropathic pain. Sprague–Dawley rats were divided into four groups, including sham, sham+KMUP‐1, CCI and CCI+KMUP‐1. KMUP‐1 (5 mg/kg, i.p.) was injected once daily starting at day 1 after surgery. Mechanical and thermal responses were assessed before surgery and at day 3, 7, 14 after CCI. Thermal hyperalgesia and mechanical allodynia were reduced in CCI+KMUP‐1 compared with CCI group. Sciatic nerves were isolated for western blots and enzyme‐linked immunosorbent assay to analyze proteins and cytokines levels. Inflammatory proteins (COX2, iNOS, nNOS) and proinflammatory mediators (IL‐1β, TNF‐α) induced by CCI were decreased in KMUP‐1‐treated group at day 7 after surgery. KMUP‐1 also inhibited neuropathic pain‐related mechanisms, including p38 and ERK activation, but not JNK, which are members of MAPKs. KMUP‐1 also blocked IκBα phosphorylation and NF‐κB translocation to nuclei. We suggest that KMUP‐1 has anti‐inflammatory and anti‐hyperalgesia properties in CCI‐induced neuropathic pain via inhibition of MAPK and NF‐κB.
Suppression of phenylephrine contraction by AMPK inhibition in isolated carotid artery from normotensive WKY and hypertensive SHR rats
Authors: Holder Peter Elliot , Rush James W E
Published: June 2021

AMP‐activated protein kinase (AMPK) is emerging as an important regulator of vascular function and has differential effects on vascular tone in arteries from normotensive Wistar Kyoto (WKY) and spontaneously hypertensive (SHR) rats. The current study aims to determine: if AMPK modulates the vasocontraction response to phenylephrine (PE); if AMPK effects are mediated by a downstream ROCK effect; and, if hypertension alters the regulatory effects of AMPK. Isolated common carotid artery (CCA) segments (denuded of endothelium) from WKY and SHR were used to determine vasomotor dose‐responses to PE (10−8–10−4 M) after control (CON) incubation, or incubation with: AMPK inhibitor Compound C (CC; 20 μM); ROCK inhibitor Y26732 (1 μM); or, CC+Y26732. PE contraction was suppressed in SHR vs WKY CON (EC50 55±6 vs 17 ± 4 nM, P <0.05). CC or Y26732 each suppressed PE contraction (increased EC50 vs respective CON) in CCA of each of WKY and SHR. The CC + Y26732 effect was greater than either individual drug effect in both WKY and SHR, but was not completely additive. While the absolute values of EC50 were higher in SHR vs WKY for each condition, the effects of CC and Y26732 on PE EC50 were relatively (to respective CON) greater in WKY vs SHR. Results suggest that AMPK may contribute to modulating PE contraction in denuded CCA via Rock‐dependent and –independent mechanisms, and that hypertension suppresses this effect of AMPK. Supported by NSERC
Genetically Engineered Mesenchymal Stem Cells that Overexpress ACE2 or Angiotensin‐(1–7) Show Enhanced Nitric‐Oxide Production
Authors: Shenoy Vinayak , Liu Meng , Rathinasabapathy Anandharajan , Qi Yanfei , Katovich Michael J , Raizada Mohan K
Published: June 2021

Administration of MSCs has been proposed for the treatment of cardiopulmonary diseases. This study was undertaken to determine whether overexpression of Angiotensin converting enzyme2 (ACE2) or Angiotensin‐(1–7) [Ang‐(1–7)] could enhance the production of nitric oxide (NO). MSCs were isolated from the femur and tibial bones of male SD rats. The cultured cells tested positive for the surface markers CD44 and CD90, and negative for CD34 and CD45 by flow cytometry. Also, they differentiated into adipocytes, confirming their mesenchymal stem cell origin. Passage‐3 MSCs were engineered to overexpress ACE2, Ang‐(1–7) or GFP following lentiviral vector‐mediated transduction. Quantitative real time PCR showed significant elevation in ACE2 levels in infected cells. Also, enzymatic assays and ELISA confirmed increased activity of ACE2 and secreted levels of Ang‐(1–7) respectively. After 72hrs of infection, the bathing medium was collected to determine NO levels using the Griess reagent system. NO production was significantly increased in MSCs overexpressing ACE2 and Ang‐(1–7) as compared with GFP or uninfected cells ( Uninfected MSCs: 4.6+0.7μM; GFP‐MSCs: 3.7+0.3μM; ACE2‐MSCs: 6.7+0.3μM Ang‐(1–7)‐MSCs: 8+0.5μM; p<0.05). These results suggest that ACE2/Ang‐(1–7) stimulates the basal production of NO in MSCs, which could enhance their therapeutic potential for the treatment of cardiopulmonary diseases.
Renal denervation prevents renal T cell activation in mice during angiotensin II‐induced hypertension
Authors: Xiao Liang , Zhang Feng , Kirabo Annet , Harrison David G
Published: June 2021

Increased sympathetic outflow has been implicated in the pathogenesis of hypertension. Recent evidence has also suggested a role of T lymphocytes in hypertension. The link between the central nervous system and immune activation in hypertension is however unclear. We hypothesized that renal sympathetic nerves activate T cells in the kidney in hypertension. We performed unilateral renal denervation in C57BL/6 mice by applying phenol to the left renal artery and cutting large visible nerves. On the same day mice received an osmotic minipump for angiotensin II infusion. Denervation decreased renal norepinephrine content from 162±13 to 42±11 ng/g (p < 0.05) as measured by HPLC. Analysis of T cells was performed using flow cytometry in digested kidneys. T cell (CD3+) number was 1.57±0.23×105 in the innervated kidney of sham infused mice and this was doubled by Ang II infusion (3.50±0.89×105). Unilateral renal denervation prevented this increase (1.60±0.31×105vs. intact side, p<0.05) from the same animals. This was accompanied by reduction in the T cell memory marker CD44 and T cell IFN‐γ production. These results suggest a potential role of renal sympathetic nerve traffic linking central command and peripheral inflammation in hypertension. Our data further show that renal sympathectomy might have anti‐inflammatory roles contributing to blood pressure control.
Cardiorespiratory responses to hypercapnia in chickens after embryonic exposure to CO2
Authors: Espinha Livia P , de Souza Fernando A , Bícego Kenia C , Macari Marcos , Gargaglioni Luciane H.
Published: June 2021

We evaluated cardiorespiratory responses to 7% CO2 of 10 (d10) and 21 (d21) days old broiler males (M) and females (F) after embryonic hypercapnia (1.5% CO2) and control situation (CS). Under normocapnia no difference in ventilation (VE), tidal volume, respiratory frequency, mean arterial pressure (MAP) and heart rate (HR) was observed between groups. Hypercapnia promoted a higher VE in Fd10 and Fd21 incubated in 1.5%CO2 compared with FCS (Fd10 control: 1497±213.4 vs Fd10 1.5% CO2: 1834±111.6; Fd21 control: 179±36.24 vs Fd21 1.5% CO2: 389±86.4 mLkg−1min− 1). Only Md21 incubated with 1.5% CO2 presented higher VE compared with CS (Md10 control: 1728±289.6 vs Md10 1.5% CO2: 1823±209; Md21 control: 239±40.4 vs Md21 1.5% CO2: 388±77.5 mLkg−1min−1). MAP was not different between groups, however, HR was higher in Fd10 incubated in 1.5% CO2 compared to Fd10 incubated in CS during hypercapnic exposure (Fd10 control: 351±25.1 vs Fd10 1.5% CO2: 405±14.8; Fd21 control: 324±11.8 vs Fd21 1.5% CO2: 331± 19.6 beats.min−1). Therefore, cardiorespiratory responses to hypercapnia are higher in animals incubated with 1.5% CO2 compared to control animals and these responses are more pronounced in females. Financial Support: FAPESP, CNPq and INCT of Comparative Physiology.
Enteric Glial Cells Activate Colon Cancer Stem Cells to Promote Tumorigenesis
Authors: Valès Simon , Biraud Mandy , Marionneau‐Lambot Séverine , Oullier Thibauld , Neunlist Michel , Van Landeghem Laurianne
Published: June 2021

Colon cancer stem cells (CSCs) are regulated by their cellular microenvironment that delivers paracrine signals crucial to CSC maintenance and tumor growth. Among CSC neighboring cells are enteric glial cells (EGCs) that are potent regulators of intestinal epithelium functions, but whose impact on CSCs has never been studied. We hypothesized that EGCs modulate CSC functions and associated tumorigenesis.

In vitro. CSCs were FACS‐isolated from HT29 colon cancer epithelial cells (CD44High/CD133High) and 3D cultured in Matrigel in presence of EGCs seeded on Transwell filters. Impact of EGCs was assessed on numbers and size of tumorspheres grown from CSCs and compared to known EGC‐derived soluble factors. In vivo. CSCs were injected alone vs. concomitantly with EGCs subcutaneously in immunodeficient mice.

In vitro EGCs increased numbers and size of tumorspheres grown from CSCs. In vivo concomitant injection of CSCs and EGCs increased tumor load vs. CSCs alone. In vitro EGC‐conditioned medium (CM) did not impact CSCs, but CM of EGCs that had been pre‐incubated with HT29 cells increased tumorsphere numbers, indicating that tumor cells activate EGCs to acquire pro‐tumorigenic abilities. Among all known glial factors tested, only prostaglandin E2 (PGE2) reproduced EGC effects on CSCs. HT29 cells increased expression of mPGES‐1 (enzyme generating PGE2) and PGE2 release in EGCs. CAY10526 (mPGES‐1 inhibitor) abolished pro‐tumorigenic properties induced by HT29 cells in EGCs.

These results suggest that tumor cells activate EGCs to acquire pro‐tumorigenic abilities, and that tumor‐activated EGCs stimulate CSC tumorigenicity via PGE2‐dependent pathways.
Stable ASCT2 and LAT1 Suppression in Human Hepatocellular Carcinoma Cells Fails to Measurably Affect Proliferation, Anchorage‐independent Growth and mTORC1 Signaling
Authors: Bothwell Paige , Wittke Evan , Papa Afreen , Mitzelfelt Jordon , Howard Zachary , Kron Clare , Bode Barrie
Published: June 2021

Amino acid transporters ASCT2 and LAT1 are coordinately enhanced in human cancers where, among other roles, they are thought to drive mammalian target‐of‐rapamycin (mTOR) growth signaling. To assess their value as therapeutic targets in hepatocellular carcinoma cells (HCC), both transporters were targeted and suppressed with lentiviral vectors producing specific short hairpin RNA in SK‐Hep1 cells. Five ASCT2 and four LAT1 stable knockdown cell lines were produced (each with a unique shRNA sequence) with proportionally suppressed glutamine and leucine transport, respectively, and suppressed transporter expression at the mRNA and protein level by 20% to 50% relative to controls. Suppression of each transporter failed to affect cell proliferation. In addition, soft agar colony formation was not compromised in transporter‐suppressed cells; neither was basal or amino acid‐stimulated mTOR complex 1 (mTORC1) signaling as assessed by 4E‐BP1 phosphorylation. No evidence was found for an enhanced reliance on autophagy or compensation by other transporters in ASCT2‐ or LAT1‐suppressed cells. Given that past ASCT2 silencing studies with inducible antisense or siRNA led to HCC apoptotic cell death, it appears that cancer cells adapt to a threshold level of suppression for these two transporter “partners in crime”. Studies with siRNA targeting both transporters are currently underway to test this “transporter threshold expression” model. These results demonstrate the adaptive plasticity of cancer cells to nutrient transporter targeted therapies, and highlight the importance of comprehensive silencing. Support: NIH Grant# R15CA108519 (BPB)
The effect of valproic acid on notch signaling in Danio rerio intestinal cell differentiation
Authors: Llorente Kasey , Uno Jennifer
Published: June 2021

Inflammatory bowel disease (IBD) is a classification of chronic remittent immune disorders affecting approximately 1.4 million people in the United States between the ages of 15 and 40. It is thought to be caused by an interaction between genetic predisposition and environmental factors, an exact mechanism explaining the onset of IBD remains unclear. Recent studies indicate a reduced number of mucus secreting cells is typical in affected individuals, and that the differentiation of intestinal cell types into either absorptive or secretory is governed by a Notch signaling pathway. There remain gaps in knowledge of how this pathway is activated or inhibited in people with chronic IBD. The purpose of this study was to induce the Notch pathway in zebrafish. We hypothesize that a corollary shift in the expression of secretory versus absorptive cell types in the gut would be observed in fish treated with valproic acid (VPA). Studies showed that VPA, a compound frequently used in the treatment of neurological disorders, activates Notch in the regulation of neuronal cells. A dosage curve was generated and the optimal concentration of VPA to administer that appeared to have an effect, without overdosing the zebrafish was determined to be 500 and 1000 µM. Following VPA treatment, brain and intestinal tissue was extracted and an RT‐PCR was run. A 30% increase in the expression of the Notch receptor was observed in brain tissue indicating VPA maybe activating Notch signaling. Preliminary results indicate that zebrafish are a viable model to study Notch signaling in the intestine. Future experiments will focus on intestinal RT‐PCR to confirm Notch receptor expression and western blot analysis will be conducted to observe any differences in the amount of secretory cells and enterocytes in the intestines following VPA treatment.
Enteral Obestatin and Ghrelin Influences on Intestinal Contractility in Piglets ‐ in vitro Studies
Authors: Woliński Jarosław , Słupecka Monika , Ochniewicz Piotr
Published: June 2021

Ghrelin and Obestatin are peptides encoded by the same ghrelin gene. Substantial amount of both peptides has been found in colostrum and milk. We have investigated the intestinal contractility of suckling piglets (7 days old, n = 10) fed every 75minutes by artificial sow system in laboratory condition enterally treated (3 x per day) with obestatin (10μg/kg b.wt. O), ghrelin (10μg/kg b.wt. G) or saline (C), (n=6 for each group), for 7 days. Duodenal and middle jejunum whole‐thickness preparations from piglets (21days old) were studied in an organ bath, for isometric recording under treatment with acetylocholine (ACh), atropine and tetradotoxin (TTX). The electrical field stimulation (EFS) was performed (90 V, duration 10 sec) at three frequencies: 0.5, 5 and 50Hz with 1 min intervals. Significant decrease in amplitude was observed after treatment with both peptides for EFS–stimulated as well as spontaneous contractile activity in duodenum. No changes were observed in jejunum. In obestatin and ghrelin treated animals injection of atropine did not result in a significant decrease of amplitude of spontaneous contractility of both intestinal segments. In all experimental groups the amplitude of EFS – stimulated and spontaneous contractility was sensitive to treatment with TTX. Our results indicate the importance of peripheral obestatin and ghrelin as regulators of intestinal contractility in piglets during early postnatal period.

Project of Polish National Science Centre 2012/05/B/NZ9/00901
Enteric Nervous System Transduction Following IV Delivery of Multiple AAV Serotypes
Authors: Gombash Lampe Sara , Cowley Christopher , Fitzgerald Julie , Mueller Christian , Christofi Fedias , Foust Kevin
Published: June 2021

Gene therapy may be useful to treat and study the enteric nervous system (ENS). In this study, we characterized transduction efficiency and distribution in the myenteric and submucosal plexuses following intravenous (IV) injection of adeno‐associated viral vectors (AAV) expressing GFP in mice. Neonate (P1) mice were injected with AAV9‐CB‐GFP (3 x 1011 vg) in the temporal vein and juvenile mice (P21) received tail vein injections (2 x 1012 vg). Mice were euthanized at 60‐90 days of age, and the myenteric plexus was examined. The myenteric plexus had GFP positive neurons along the entire GI tract in injected mice. Segmental examinations of myenteric neurons in the colon showed no oral‐aboral bias. Neuronal transduction ranged from 21‐57% depending on the region. GFP co‐localized with interneurons, excitatory and sensory myenteric neurons. In contrast, GFP expression in VIP, nNOS and glial cells was absent. However, IV injection of AAV9 with a glial promoter (GFAP) drove GFP expression exclusively in glia. Neonate IV injections with AAV1, 5, and 6 containing the CB‐GFP expression cassette produced little to no transgene expression in the myenteric plexus. However, AAV8‐CB‐GFP showed significantly higher neuronal transduction compared to AAV9 injected mice. AAV8‐CB‐GFP expressed in all ENS cell types examined including VIP and nNOS neurons and intraganglionic S100 glia. Characterization of AAV‐CB‐GFP transduction in submucosal neurons is ongoing. These data demonstrate that the ENS is efficiently targeted by AAV8 and AAV9 following IV delivery. Further, transgene expression can be customized with changes in the viral capsid and expression cassette.
Gintonin‐Mediated Depolarization of Pacemaker Activity in Cultured Interstitial Cells of Cajal
Authors: Kim Byung Joo , Nam Joo Hyun , Joo Myungsoo , Ha Tal Soo , Choi Seok , Jun Jae Yeoul , So Insuk , Nah Seung‐Yeol
Published: June 2021

We investigated the effect of gintonin, a novel ginseng‐derived G protein‐coupled lysophosphatidic acid (LPA) receptor ligand, on the pacemaker activity of the interstitial cells of Cajal (ICC) in murine small intestine and GI motility. The whole‐cell patch‐clamp configuration was used to record pacemaker potentials and currents from cultured ICC in the absence or presence of gintonin. In vivo effects of gintonin on GI motility were investigated by measuring the intestinal transit rate (ITR) of Evans blue in normal and streptozotocin (STZ)‐induced diabetic mice. Gintonin caused membrane depolarization but this action was blocked by Ki16425, an LPA1/3 receptor antagonist. Next, we examined the effects of U‐73122, an active PLC inhibitor, and chelerythrine and calphostin, which inhibit PKC. All inhibitors blocked gintonin actions on pacemaker potentials, but not completely. Gintonin‐mediated depolarization was lower in Ca2+‐free than in Ca2+‐containing external solutions and was blocked by thapsigargin. We found that gintonin also activated Ca2+‐activated Cl‐ channels (ANO1), but not TRPM7 channels. In vivo, gintonin (10‐100 mg/kg), not only significantly increased the ITR in normal mice but also ameliorated STZ‐induced diabetic GI motility retardation. Gintonin could be a novel candidate for development as a prokinetic agent that may prevent or alleviate GI motility dysfunctions in human patients.
The Zinc Transporter ZnT2 is Necessary for Stabilization of Granule Contents in Intestinal Paneth Cell Granules
Authors: Podany Abigail , Geletzke Abby , Lee Sooyeon , Soybel David , Kelleher Shannon
Published: June 2021

Paneth cells (PC) are secretory cells in small intestinal crypts that play a critical role in self‐protection against luminal pathogens. Defects in PC function are associated with necrotizing enterocolitis and Crohn's disease. PCs release granules containing a host of antimicrobial peptides and a large amount of zinc (Zn). Zn prevents degradation of α ‐defensins and stabilizes lysozyme in its monomeric active conformation. How Zn accumulates in PC granules is not understood. We used wild‐type (WT) and ZnT2‐null (KO) mice to determine the role of ZnT2 in PC function. ZnT2 was expressed in crypts and localized to PC granules by immunofluorescence and sucrose gradient fractionation. Studies using the fluorescent Zn reporter ZP1 indicated that KO mice were unable to accumulate Zn into granules. While no morphologic abnormalities were observed in the intestines of KO mice, a significantly greater number of PCs were degranulated compared with PCs in WT mice (p<0.01), and PCs in KO mice had profound defects in sub‐cellular organization. In response to saline perfusion, ileal segments from KO mice retained less overall (p<0.01), and significantly more dimerized (less active) lysozyme (p<0.05), but released more lysozyme into the lumen (p<0.05). These data indicate that ZnT2 imports Zn into PC granules, and the inability to do so destabilizes granules and Zn‐requiring granule proteins and leads to defects in coordinated granule secretion, which may put an individual at risk for infection and inflammatory illness.
An in situ model of chronic cervical spinal cord injury
Authors: Baekey David , Amirzadehasl Parisa , Armstrong Gregory , Fuller David , Reier Paul
Published: June 2021

In vivo rodent models of cervical spinal cord injury (cSCI) are commonly used to examine impaired respiratory activity using measures of either phrenic nerve output or diaphragmatic activity. These methods allow quantification of overall deficits due to hemisection or contusion injury but hemostatic and ventilatory movement issues often confound recordings of affected neural circuitry. To address this challenge, we have developed an in situ rodent model of cSCI. This preparation uses an established method of contusion injury (Infinite Horizon pneumatic impactor – 100 kdyne at C4) in juvenile (P21) rodents and allows up to 2 weeks of recovery prior to experimental recording. The improved accessibility and elimination of hemostatic concerns facilitates recording of individual cervical and thoracic ventral roots while allowing stable monitoring of neuronal ensembles using multielectrode array technology. Additionally, this decerebrate and artificially perfused preparation removes the need for anesthesia. Initial recordings demonstrate segmental efferent impairment (C4 and C5) corresponding to injury location, with sparse inspiratory effort representing a reduction of motoneuron activity. Increased tonic and expiratory activity of inspiratory ventral roots contributing to the phrenic nerve (ipsilateral to injury) suggest dysfunction of the local network. We propose that this model is a significant improvement over in vivo paradigms for defining both the spinal circuitry and plasticity involved in recovery from cSCI. Funded by the University of Florida College of Veterinary Medicine, NIH 1R01NS080180‐01A1 and R01 NS054025.
GAL‐160 Preferentially Increases Respiratory Motor Drive to the Tongue over the Diaphragm during Unobstructed Breathing and Standardized Obstructive Apneas (OA) in Rats
Authors: Baby S , Mardirosian S , Schemm C , Peng S , Golder F , MacIntyre E
Published: June 2021

GAL‐160, a carotid body (CB) stimulant, decreases spontaneous OA frequency and OA duration in a rodent model of OSA. Mechanistically, we propose that the effect of GAL‐160 on OA frequency is due to increased upper airway respiratory muscle (RMUA) activity during eupnea, and the effect of GAL‐160 on OA duration is due to an increased response of RMUA to obstruction. We evaluated the effects of GAL‐160 on eupneic respiratory drive to the RMUA by recording hypoglossal (HN) and phrenic (PN) neurograms from urethane anesthetized, mechanically ventilated rats before and during IV infusion of vehicle and GAL‐160 (0.24 mg/kg over 30‐min). Baseline neurograms were standardized between rats by setting baseline PaCO2 at 3 mmHg above each animal's apneic threshold to CO2. To evaluate the effects of GAL‐160 on the RMUA response to obstruction, the EMGGG and EMGDIA responses to standardized 10‐s tracheal obstructions were recorded from anesthetized, spontaneously breathing rats, before and during GAL‐160/vehicle infusions.

GAL‐160 increased HN activity (67 ± 23% above baseline) without altering PN activity (‐13 ± 20%) during unobstructed breathing. Bilateral CB denervation abolished this effect. GAL‐160 increased the peak EMGGG response to a 10‐s obstruction (pre‐drug: 214 ± 180%; GAL‐160: 589 ± 358%; p<0.01) without altering the EMGDIA response (pre‐drug: 60 ± 31%; GAL‐160: 78 ± 47%). Vehicle infusion had no effect on the neurograms or EMGs recorded. These data demonstrate that GAL‐160 preferentially increases respiratory drive to the RMUA via the CB and during both eupneic breathing and OA. This mechanism may contribute to the efficacy of GAL‐160 in our rodent model of spontaneous OSA.
Alveolar Overdistension Does Not Occur Even at Very High Airway Pressure
Authors: Jain Sumeet , Kollisch‐Singule Michaela , Gatto Louis , Nieman Gary , Habashi Nader
Published: June 2021

Objective

Acute respiratory distress syndrome (ARDS) is a disease with unacceptably high morbidity and mortality. Improper mechanical ventilation can cause a secondary ventilator induced lung injury (VILI), which significantly increases ARDS mortality and incidence. VILI is thought to occur via two mechanisms: alveolar recruitment‐derecruitment (R/D) and alveolar overdistension. We hypothesize that overdistension does not occur at the alveolar level with high airway pressures in either normal or surfactant deactivated lungs.

Methods

Sprague‐Dawley rats were anesthetized, instrumented, and ventilated with continuous positive airway pressure (CPAP) ventilation with pressure increased from 5 to 10, 20, 30, 40, and 50 cmH2O. In vivo microscopy was used to photograph subpleural alveoli at each pressure level. Surfactant was deactivated using Tween lavage, and subpleural alveoli photographed at the same airway pressures. Alveolar area/photomicrograph was measured using computer image analysis.

image

(Alveoli colored in yellow)

Results

Alveolar diameter did not increase significantly between CPAP 30 and 50 cmH2O and never exceeded known normal range (63.2µm maximum average diameter) for rat lung alveoli (range 43‐102µm).

Conclusions

Subpleural alveoli do not overdistend even at high airway pressure in normal or surfactant deficient lungs. Thus gross barotrauma must occur in the conducting airways and pulmonary parenchymal injury caused by VILI must be secondary to alveolar R/D.
Ethanol Affects the Progression of Planarian Head Regeneration in a Time‐Dependent Manner
Authors: Hanna Lydia , McGee James , Srinivasan Dayalan , Staehle Mary
Published: June 2021

In humans, ethanol exposure in utero affects neurodevelopmental processes, which can lead to a spectrum of physical and cognitive impairments known as fetal alcohol spectrum disorders (FASD). The molecular etiology of FASD is poorly understood and challenging to study in most animal models. Therefore, we have established head regeneration in the Schmidtea mediterranea planarian as a model system for investigating the direct effects of ethanol exposure on neurodevelopmental processes. Our previous work elucidated delayed reacquisition of cognitive behavior in 1% v/v alcohol‐exposed planaria following head removal. In this study, we used a limited exposure paradigm to define the developmental window(s) sensitive to alcohol exposure. Phenotypic and morphological changes were monitored with cognitive function testing (a movement‐normalized photophobic behavior assay) and whole‐mount immunostaining of the nervous system. The behavioral data suggest that alcohol's effects occur between 36 and 120 hours after transection; delayed reacquisition of photophobic behavior occurs only when the planaria are exposed to alcohol for 48‐hour periods within this time frame. Delayed development is also observed in the nervous system morphology of alcohol‐exposed worm. Analyzing the underlying molecular mechanisms of neurodevelopment during this developmental period could elucidate alcohol's effects on neural stem cell differentiation and the development of FASD.
Adolescent Delta‐9‐Tetrahydrocannabinol (THC) Pre‐Exposure Fails to Attenuate Adulthood Place and Taste Avoidance
Authors: Wakeford Alison , Flax Shaun , Pomfrey Rebecca , Riley Anthony
Published: June 2021

Adolescent initiation of drug use has been linked to problematic drug consumption later on in life, and may represent an important variable in the shifting of rewarding and/or aversive properties of drugs of abuse. The current study examined adolescent pre‐exposure of THC to adulthood conditioning to THC in a combined place and taste conditioning apparatus. Adolescents received eight pre‐exposure injections to an intermediate dose of THC (3.2 mg/kg) or vehicle from PND 21‐PND 50 (one every third day). After pre‐exposure, animals were tested in a combined CTA/CPP procedure in adulthood (PND ~90). Animals were exposed to 20 minutes of a novel tastant, immediately injected with a dose of THC (1.0, 5.0 mg/kg IP) or vehicle, and then confined to one side of the place conditioning apparatus. On the following day, animals received access to 20 minutes of water, immediately followed by an injection of vehicle, and placed in the other side of the chamber (non‐drug paired side). This schedule of one conditioning day followed by one water recovery day is considered one cycle. Animals were exposed to four cycles, a final CPP test, and then a one‐bottle avoidance test. Adolescent pre exposure had no effect on adult saccharin consumption, or place conditioning to THC. THC induced dose dependent avoidance in taste conditioning, and avoidance at the highest dose in place conditioning. Results will be discussed regarding preclinical models that may further elucidate translational research between apparent rodent avoidance of THC and human preference to cannabis. Research supported by the Mellon Foundation to ALR.
Evaluation of FAAH‐ and MGL‐Inhibition in Cannabinoid Drug Discrimination
Authors: Leonard Michael , Kangas Brian , Makriyannis Alexandros , Nikas Spyros , Shukla V. , Alapafuja Shakiru , Bergman Jack
Published: June 2021

The discriminative‐stimulus (SD) effects of CB1 ligands can be reproduced in mice by non‐selective inhibition of the anandamide‐(AEA‐) and 2‐arachidinoyl glycerol‐(2‐AG‐) inactivating enzymes, respectively, fatty‐acid amide hydrolase (FAAH) and monoacyl glycerol lipase (MGL). Additionally, we have shown that nonselective FAAH/MGL inhibitors produce a CB1‐like SD effect in squirrel monkeys. The present studies with selective and nonselective ligands [FAAH (URB597; AM2018), MGL (AM2036); and FAAH/MGL‐mixed (AM2129)] were conducted to further evaluate the effects of FAAH and MGL inhibition in squirrel monkeys trained to discriminate the CB1 agonist AM4054 from vehicle (n=4). Results indicate that: 1) co‐administration of selective FAAH‐ and MGL‐inhibitors, but not FAAH‐ or MGL‐selective inhibitors alone, produces CB1‐related SD effects;2) AEA, but not 2‐AG, produces CB1‐like SD effects after either FAAH or mixed FAAH/MGL inhibition; and 3) CB1 receptor antagonism effectively and blocks the CB1‐like effects of combined FAAH/MGL inhibition; this effect was insurmountable. The present data strengthen the idea that complementary, but not individual, actions of the endocannabinoids AEA and 2‐AG produce CB1‐related SD, and perhaps subjective, effects in primate species.
Chronic‐Binge Alcohol‐Induced Hepatic Injury and Inflammation Were Ameliorated in Mice Deficient for Transient Receptor Potential Vanilloid 1 Gene
Authors: Kirpich Irina , Falkner Keith , Beier Juliane , Arteel Gavin , McClain Craig
Published: June 2021

Introduction

Experimental alcohol‐induced liver injury is exacerbated by a high polyunsaturated fat diet rich in linoleic acid (LA). The aim of the study was to evaluate the role of bioactive oxidized LA metabolites (OXLAMs) and their signaling through the Transient Receptor Potential Vanilloid 1 (TRPV1) in an experimental animal model of alcoholic liver disease (ALD).

Methods

C57BL/6 wild type (WT) and Trpv1 knock out mice were fed a diet containing 5% ethanol for 10 days, followed by a single ethanol gavage. Liver steatosis, injury and inflammation were assessed. In vitro studies using HepG2 cells were performed to evaluate OXLAM/TRPV1 signaling.

Results

Chronic‐binge alcohol administration to WT animals resulted in a marked increase in plasma OXLAM levels, specifically 9‐ and 13‐HODEs, in parallel with the up‐regulation of hepatic Trpv1. These effects were associated with hepatic steatosis, inflammation and injury. Genetic depletion of Trpv1 did not blunt hepatic steatosis caused by EtOH, but ameliorated hepatic injury and prevented the increase in pro‐inflammatory cytokine and chemokine expression, including Tnf‐α, IL‐6, Mip‐2, Mcp1, and Pai‐1. Exposure of HepG2 cells to 9‐ and 13‐HODEs resulted in activation of TRPV1 signal transduction with the increased intracellular Ca2+ levels, suggesting that OXLAM/TRPV1/Ca2+ signaling may be a relevant pathway contributing to ALD.

Conclusions

TRPV1‐OXLAM interactions appear to play a significant role in ethanol‐induced hepatic inflammation/injury, further supporting an important role for dietary lipids in ALD.
Characterization of Neurons Derived from a Human Induced Pluripotent Stem Cell Line Using Patch‐Clamp Electrophysiology
Authors: Metz Kathleen , Cao William , Coyne Leanne , Halliwell Robert
Published: June 2021

Neuropharmacological studies using animal‐derived tissues are not always directly translatable to humans, and the occurrence of discrepancies can be difficult to predict. Sources of human neurons have been limited in the past, but developments in stem cell science are now providing new opportunities to generate human neurons from stem cells. iCell neurons are derived from a human induced pluripotent stem cell (hIPSC) line and may have the potential to replace animal neurons in neuropharmacological studies. Here, we have explored the neurophysiological and neuropharmacological properties of iCell neurons using immunocytochemistry and single‐cell patch‐clamp techniques.

Whole cell recordings were obtained from iCell neurons cultured for up to 4 months in vitro. Functional expression of voltage‐gated sodium and potassium ion channels, as well as ligand‐gated GABAA, glutamate, NMDA, AMPA, kainate and glycine receptors was determined. Spontaneous and evoked action potentials were inhibited by the sodium channel blocker tetrodotoxin, and spontaneous activity levels increased over time in culture. Immunocytochemical analysis revealed a corresponding increase in neurite density over time. iCell neurons therefore appear to have several important functional properties consistent with human neurons and might be a viable alternative to animal neurons in drug studies.

Research support was provided by Cellular Dynamics International and the Department of Physiology and Pharmacology at the University of the Pacific.