bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of Omega‐6 HUFA Status on Recovery of Muscle Performance After Fatiguing Exercise
Authors: Montain Scott , Johannsen Neil , Champagne Catherine , Marriott Bernadette , Hibbeln Joseph , Hawes Michael , Berry Kevin
Published: June 2021

High %n‐6 HUFA in plasma and tissues is associated with a heightened inflammatory response.

PURPOSE

To test if a diet that lowers the %n‐6 would be associated with less strength loss after fatiguing load carriage exercise and shorter time to recover muscle strength and performance.

METHODS

Volunteers were randomly assigned to either a control diet (CON; n=23; 22.5 g n‐6 and 0.2 g n‐3 HUFA/2,000 kcal) plus placebo smoothie (‐S), an experimental diet (EXP; n=25; 8.0 g n‐6 and 0.9 g n‐3 HUFA/2,000 kcal) and ‐S, or EXP plus n‐3 fortified smoothie (EXP+S; n=23; 1.9 g n‐3 HUFA) for 10 weeks. Volunteers performed a single leg step test (up to 3 sets & 6 min each leg) while wearing a 20 kg vest. Maximal isometric and isokinetic torque at 60°/sec, and total work during 20 reps at 180°/sec were measured before, 5 min after the step test, and 24, 48 and 72 h of recovery.

RESULTS

The step test produced a 20% (95%CI 16‐25), 19% (12‐25), and 19% (15‐27) reduction in isometric torque for EXP+S, EXP‐S, and CON, respectively, with no group x time interaction during recovery (p=0.89). A similar magnitude and pattern was observed for isokinetic torque and total work.

CONCLUSION

A 10‐wk dietary manipulation that substantially lowered %n‐6 and increased %n‐3 HUFA was not associated with less fatigue or faster recovery after load carriage exercise. The views expressed in this abstract are those of the authors and do not reflect the official policy of the Dept. Army, Dept. Defense, or the U.S. Government.
Safety of Alternate Day Fasting and Effect on Disordered Eating Behaviors
Authors: Hoddy Kristin , Kroeger Cynthia , Trepanowski John , Barnosky Adrienne , Bhutani Surabhi , Varady Krista
Published: June 2021

Alternate day fasting (ADF; ad libitum intake “feed day” alternated with 75% restriction “fast day”), is effective for weight loss, but the safety of the diet has been questioned. Accordingly, this study examined occurrences of adverse events and eating disorder symptoms during ADF. Obese subjects (n = 59) participated in an 8‐week ADF protocol where food was provided on the fast day. Body weight decreased (P < 0.0001) by 4.2 ± 0.3%. Some subjects reported constipation (17%), water retention (2%), dizziness (<20%), and general weakness (<15%). Bad breath doubled from the first week of the diet (14%) to the last week (29%), though not significantly. Depression and binge eating decreased (P < 0.01) with ADF. Purgative behavior and fear of fatness remained unchanged. ADF helped subjects increase (P < 0.01) restrictive eating and improve (P < 0.01) body image perception. Therefore, ADF produces minimal adverse outcomes, and has either benign or beneficial effects on eating disorder symptoms.
Anti‐obesity Effect of Solar Salt Produced in Korea on C57BL/6 Mice
Authors: Park Kun‐Young , Ju Jae‐Hyun , Song Jia‐Le , Mun Suk‐Hee
Published: June 2021

The mineral contents of several solar salts produced in South Korea and the anti‐obesity effect of the solar salt on C57BL/6 mice induced by HFD were studied. We used purified salt (PS), Korean solar salts from Taepyeong salt pond (SS‐T), Yangchunsa (SS‐Y), Shinil‐1 (SS‐S1), Shinil‐2 (SS‐S2), and French Guerande solar salt (SS‐G) as a positive sample. The mineral contents of those samples were determined by ICP‐OES. Of all samples, SS‐T had the highest concentration of calcium (0.2388%) and sulfur (1.0313%), and SS‐S1 had the lowest Na/K ratio (17.81), all of which are superior to those of SS‐G. Also, all of salt samples were free from hazardous heavy metals. We used C57BL/6 mice for high‐fat‐diet‐induced obesity; 8‐week obesity induction using high‐fat diet (HFD), followed by 8‐week treatment of HFD supplemented with 0.47% of salt including PS, SS‐T, SS‐Y, SS‐S1, SS‐S2, and SS‐G. Compared with SS‐G, SS‐T and SS‐S1 effectively reduced body weight gain, food efficiency ratio, serum triacylglycerol, cholesterol and leptin. In addition, SS‐S1 significantly reduced organ weights such as epididymal adipose tissue and liver in comparison with control and SS‐G. Also, SS‐S1 was found to suppress adipogenesis and induce lipolysis in histochemical analyses with hepatic‐ and epididymal adipose tissues. And SS‐S1 significantly suppressed expression of obesity‐related genes such as PPAR‐γ, FAS and CPT‐1. Taken together, administration of solar salt, especially manufactured in Korea, can suppress progression of obesity, probably due to relatively high concentration of beneficial minerals.
Dietary fat intake and age influence gut microbiota and colonic inflammation in C57BL/6J mice
Authors: Kim Su Jeong , Sung Mi‐Kyung , Park Mi‐Young , Kim Sun‐Eun
Published: June 2021

Recent studies have showed that diet‐induced alterations of gut microbiota composition play a pivotal role in the development of metabolic diseases. We investigated whether dietary fat and age would affect gut microbiota, permeability and inflammation. C57BL/6J mice were randomly assigned to either normal fat diet (ND) or high‐fat diet (HD) group. After 10 wks, a half of mice in each group were switched to either HD or ND feeding for additional 10 wks. Microbiome composition and diversity were analyzed by 16S rRNA‐based pyrosequencing. Colonic mRNA expressions of tight junction (TJ) proteins and inflammatory cytokines were measured by qPCR. DNA strand break was determined using comet assay. The main bacterial phyla of mice were Frimicutes, Bacteroidetes, and Actinobacteria. Diversity of gut microbiota was reduced in mice fed HD compared to those of mice fed ND. In mice fed HD for 20 wks, the proportions of Actinobacteria and Firmicutes increased while the proportion of Bacteroidetes decreased compared to mice fed ND for 20wks. The proportions of Firmicutes and Bacteroidetes were related with age while the proportion of Actinobacteria was related with dietary fat content. High‐fat diet was negatively associated with the expressions of TJ proteins while it was positively associated with the expression of inflammatory cytokines. Changes in the expression of TJ protein and inflammatory cytokines followed changes in fat content of the diet. Our data showed that both dietary fat intake and age affect gut microbiota composition as well as colonic membrane integrity and inflammation. [This study was supported by the Mid‐Career Research Program 2012R1A2A2A01046228 NRF]
Diet Associations With Sleep and Fatigue Among Truck Drivers: Baseline Results From the SHIFT Randomized Controlled Trial
Authors: Thompson Sharon , Hohn Elliot , Wipfli Brad , Olson Ryan
Published: June 2021

This study examined baseline associations between diet and sleep/fatigue among commercial truck drivers enrolled in a randomized controlled trial of the Safety & Health Involvement For Truckers (SHIFT) weight loss and health promotion program (NHLBI, R01HL1054950). Participants (n=454) completed a written survey and health assessment that included a 10‐minute psychomotor vigilance task (PVT). Objective sleep actigraphy was conducted for 7‐10 days with a subsample of drivers using the Actiwatch Spectrum. Diet‐focused survey measures included fruit and vegetable servings, percentage energy from fat, and intake of fast food and sugary drinks/snacks. Sleep and fatigue survey measures included self‐reported sleep quality. PVT outcomes included reciprocal response time, slow responses, and lapses in attention. Linear regression was used to explore relationships between variables. Age and BMI were used as covariates in analyses. Participants were majority male (86.6%) with a mean age of 47.8 years (SD=11.4) and BMI of 37.2 kg/m2 (SD=8.1). Drivers ate few daily fruit and vegetable servings (M=2.6, SD=2.3) and reported consuming sugary drinks, sugary snacks, and fast food 3‐4 times per week. Findings for fruit and vegetable servings were mixed, but included a significant relationship between higher fruit and vegetable consumption and fewer slow responses (β=‐.11, p=.024). A significant interaction was also found between sugary drinks and poor sleep quality (β=.11, p=.032). Preliminary results highlight the need for driver health interventions, and reveal dietary associations with poor sleep quality and indicators of fatigue.
Under Reporting of Hypertension in Obese Teenagers
Authors: Yeh Yulyu , Brogan Hartlieb Kathryn , Naar‐king Sylvie , Jen Cathrine K‐L
Published: June 2021

The high prevalence of obesity in the pediatric population has caused an increased in hypertension (HTN) rate in this population. However, the diagnostic of HTN may not be as thorough for children as for adults. This study compared the actual measured blood pressure (BP) level and reported diagnostic of HTN in teens and the relationship of teen and caregivers' (CG) BP levels. A convenience sample of 162 obese teenagers (mean age: 13.8±1.35 yrs, 67.3% girls, mean BMI 37.7±7.52kg/m2) and their caregivers (mean age: 43.1±8.2 yrs, 95.1% female, mean BMI: 40.5±11.2kg/m2) took part in this study. BPs were measured from both teens and CG in the home during a baseline study data collection using an automatic blood pressure monitor and the average of 3 measurements was used. Average BP was 121.1±10.3/73.7±8.9mmHg, 129.3±17.6/83.2±12.0 mmHg for teens and CGs respectively. According to CG report, 9 (5.6%) teens had been diagnosed with HTN. However, based on the measurement, 27.8% of the teen had SBP or DBP >95th percentile. Thirty‐seven teens with blood pressure over 95th percentile were not reported as having a diagnosis of HTN. Teen's SBP and DBP were both positively correlated with CG's SBP (r= 0.294, 0.175, p<0.05) and DBP blood pressure (r=0.231, 0.248, p<0.01). Teens with CG without HTN were less likely to have HTN than CG who have HTN (X2=4.985, p<0.26). The results suggest that there might be an under diagnosis of HTN in this population and teens with family history of HTN were more likely to develop HTN. More research on clinical screening and diagnosis of HTN in obese teenagers should be encouraged to address the issue.
Quinoa protective effects against obesity‐induced intestinal inflammation
Authors: Noratto G , Carrion‐Rabanal R , Medina G , Mencia A , Mohanty I , Gonzalez D , Murphy K
Published: June 2021

Quinoa (Chenopodium quinoa), contains polyphenols that can reach the colon and provide benefits for intestinal health. Our objective was to investigate the stability of polyphenols in quinoa varieties after cooking and potential effects of quinoa polyphenols in prevention of intestinal inflammation.

Extractable and non‐extractable polyphenols were assessed following standard analytical procedures. The obese diabetic (db/db) (obese control) mice and the lean counterparts (lean group) fed with a standard diet were compared to a db/db group fed with an isocaloric Std. diet supplemented with quinoa (quinoa group) for 8 weeks (n=5).

Results showed that among the quinoa varieties analyzed, Cherry Vanilla and Temuko represented the two highest and lowest extremes regarding the contents of extractable polyphenols while Faro and Black represented two extremes for the contents of non‐extractable polyphenols. However, these varieties did not reach statistical significance after cooking. Body weight gain was prevented by quinoa consumption (~ 84.87 ± 1.46% of obese control). mRNA levels of inflammatory cytokines IL‐1β, IL‐8, and TNF in colonic mucosal cells were similar in lean and quinoa groups, while in obese group were 2.5‐, 3.0‐, and 6.2‐fold of lean controls, respectively. The protective effects of colonic mucosal cells might be due to the polyphenols present in quinoa that also downregulated IL‐1β, IL‐8, and TNF cytokines in colonic epithelial Caco‐2 cells in vitro. Overall, quinoa consumption can prevent inflammation and promote gastrointestinal health in part due to its content of polyphenols.
A Lutein Nano‐Emulsion is more Effective than Regular Lutein in Protecting Cholesterol‐Induced Liver Injury by Increasing Lutein Bioavailability in Guinea Pigs
Authors: Murillo Gabriela , Hu Siqi , Aguilar David , Missimer Amanda , Smyth Joan , Gannon Sarah , Fernandez Maria‐Luz
Published: June 2021

Regular lutein (R‐lutein) vs lutein prepared in nano‐emulsion (NANO) (average particle diameter = 250 nm) were compared on their effects in reducing hepatic steatosis and liver inflammation in guinea pigs with diet‐induced liver injury. Twenty four guinea pigs were divided into 3 groups (8/group) and were fed for 6 weeks either a hypercholesterolemic diet (0.25g/100g dietary cholesterol) only (control) or in combination with 3 mg/d of R‐Lutein or 3 mg/d of lutein as NANO. Plasma lutein was 3 times higher in the NANO group compared to the R‐Lutein group (68.3 vs 22.7 nmol/L, respectively) while there was no detectable plasma lutein in control guinea pigs. Lutein concentrations in liver (p< 0.001), adipose (p< 0.001) and eyes (p< 0.001) were greater in the NANO, intermediate in the R‐Lutein and almost non‐existent in the control group indicating that lutein in NANO is more easily absorbed and delivered to the target tissues. Further, guinea pigs fed NANO lutein had hepatic steatosis scores of 2.04 ± 0.53 which were lower than those found in control (2.50 ± 0.45) or in guinea pigs from the R‐lutein group (2.63± 0.26) (p< 0.025). Inflammatory cytokines, Interleukin (IL)‐1 beta and Monocyte chemoattractant protein‐1 were lower in the NANO group compared to the R‐Lutein or the control groups (p<0.05). These results indicate that utilizing a nano‐delivery mechanism, results in higher lutein bioavailability and a greater protection against hepatic steatosis and inflammation
Effects of Acute UVB on Retinoid Metabolism
Authors: Gressel Katherine , Oberyszyn Tatiana , Everts Helen
Published: June 2021

Exposure to UVB light is the largest risk factor for the development of cutaneous squamous cell carcinoma, one of the most common cancers in the United States. UVB light causes changes to occur in the skin such as epidermal thickening, hyperproliferation, and alterations to epidermal differentiation. Retinoids regulate the differentiation and proliferation of the epidermis. The purpose of this study is to examine the effect of acute UVB on the expression of retinoid metabolism proteins. There is limited knowledge of how UVB may affect this pathway, and our results will allow us to gain a more complete understanding of changes in the entire retinoid metabolism pathway. We examined the expression of key retinoic acid metabolism proteins in the epidermis and hair follicle remnants 48 hours after UVB exposure in SKH‐1 mice using immunohistochemistry. UVB exposure increased the expression of retinoid synthesis, degradation, and binding proteins, and decreased the expression of the retinol storage enzyme, lectin:retinol acyltransferase (LRAT). These changes in expression suggest that acute UVB increases in the synthesis of retinoic acid in the epidermis. Many of these proteins changed their localization in the epidermis, concentrating in the more differentiated cells of the stratum granulosum and stratum corneum. This change in retinoic acid synthesis could affect the repair and normalization of UVB‐damaged epidermis. This work was supported by the Department of Human Sciences (Human Nutrition), The Molecular Carcinogenesis and Chemoprevention Program of The Ohio State University Comprehensive Cancer Center (grant P30 CA16058, National Cancer Institute) and the Center for Advanced Functional Foods Research and Entrepreneurship (CAFREE).
Influence of spice and grape pomace extracts on migraine‐related indicators in vitro
Authors: Slavin Margaret , Bourguignon Julia , Jackson Kyle , Orciga Michael‐Angelo
Published: June 2021

Calcitonin gene‐related peptide (CGRP) is well‐recognized as a pivotal contributor to the inflammatory process in migraine. Limited evidence suggests that diet may impact circulating levels of CGRP. The objective of this study was to investigate the effects of ginger, turmeric and grape pomace extracts on two cellular models of migraine: CGRP secretion from neuroendocrine CA77 cells, and calcium uptake by nerve growth factor‐differentiated PC12 cells.

Dried, ground samples were passed through a 40‐mesh sieve prior to extraction in solvents chosen for their efficiency: grape pomace (50% acetone), ginger root (methanol), and turmeric (acetone). Treatment doses, in 0.1% DMSO vehicle solvent, were based on cell viability results via methylene blue cytoxicity assay. CA77 cells were incubated with treatment for 1 h, then ELISA was used to quantify CGRP. Calcium influx was measured in PC12 cells via the GCaMP5 genetically encoded fluorescent calcium indicator, after 10 min pre‐treatment with extracts, upon stimulation with 50 mM KCl.

Results showed a statistically significant decrease in CGRP secretion following treatment with high doses of ginger (0.2 mg ginger equivalent/mL) and grape pomace (1.0 mg pomace equivalent/mL) extracts. Relative to vehicle control, CGRP secretion was decreased by 22% and 87%, respectively. A non‐statistical trend of lower calcium influx was observed in PC12 cells treated with ginger extracts. Solubility of turmeric extracts may have precluded detection of an effect. These results suggest that grape pomace and ginger extracts may have anti‐inflammatory propensity in migraine, which future studies may elucidate.
The anti‐inflammatory effects of wheat germ oil on lipopolysaccharide‐activated human monocytic (THP‐1) cells
Authors: Janthachotikun Sawanya , Peterson Sandra , Fiddler Joanna , Clarke Stephen , Stoecker Barbara , Dunford Nurhan , Smith Brenda , Lucas Edralin
Published: June 2021

Heart disease is the leading cause of death worldwide, and the underlying pathological feature is atherosclerosis. Atherosclerosis is an inflammatory disease involving various immune cells with macrophages as key agents in the disease progression. Macrophages play a vital role by secreting pro‐inflammatory cytokines including tumor necrosis factor (TNF)‐α and interleukin (IL)‐6. Dietary bioactive components are being studied for their beneficial properties. Wheat germ oil (WGO) is rich in omega‐3 fatty acids, vitamin E and policosanols which have been reported to have pharmacological effects, including anti‐inflammatory property. The present study investigated the anti‐inflammatory effects of different WGOs on cytokine production in lipopolysaccharide (LPS)‐activated THP‐1 cells. THP‐1 cells were treated with different concentrations (0‐1.5 x 10‐3%) of four WGOs prepared using different extraction methods (WGO1, WGO2, WGO3 and WGO4) and one commercial WGO (WGOC) with or without LPS (100 ng/mL) for 24 h. None of the WGOs tested induced cell death or had a negative effect on cell proliferation. All WGOs significantly decreased TNF‐α and IL‐6 production (P<0.0001), with WGOC being the most effective. In contrast, unrefined hexane extracted WGO4 induced the highest production of the anti‐inflammatory cytokine, IL‐10.These findings suggest that WGOs can simultaneously down regulate pro‐inflammatory and up‐regulate anti‐inflammatory responses and that the extraction method influences the magnitude of these effects.
Nigella sativa influences GLUT4 through the AMPK pathway
Authors: Acevedo Daniel , Varela Erika , Guerra Juan , Banu Jameela , Reyna Sara
Published: June 2021

Type 2 diabetes is a global epidemic. Increasing evidence shows that estrogen has protective effects on insulin sensitivity. Decreased estrogen levels affect adipocytes response to insulin leading to increased circulating glucose levels and insulin resistance. AMP‐activated protein kinase (AMPK) regulates cellular energy balance due to its role in glucose, lipid, and protein metabolism. Activation of AMPK results in translocation of glucose transporter 4 (GLUT4) to the plasma membrane which mobilizes glucose into the cell. The seed extract of Nigella sativa (NS) reduces elevated glucose levels. However, the molecular mechanisms involved in regulating glucose transport by NS remain elusive. We used an estrogen deficient animal model to examine whether NS and its active ingredient thymoquinone (TQ) modulate proteins involved in regulating glucose transport: AMPK, Akt, phosphorylated AMPK and Akt, and GLUT4.Sprague Dawley rats were either sham‐operated or ovariectomized and divided into control, NS and TQ fed groups. Rats were fed the respective diets for 4 months and sacrificed. The peritoneal adipose tissue was collected and proteins were analyzed. NS fed sham‐operated rats showed significant increase in AMPK phosphorylation, compared to control sham‐operated rats. Similarly, ovariectomized rats fed TQ showed an increase in AMPK phosphorylation, compared to TQ sham‐operated rats. Moreover, rats fed NS or TQ showed an increase in GLUT4 protein content, compared to the respective control groups. However, NS and TQ did not activate Akt phosphorylation.
Lower Weight Gain and Plasma and Liver Lipids in DIO Mice Fed Whole Grape Seed Flour Are Associated with Decreased Adipose Inflammatory Pathway Gene Expression
Authors: Kim Hyunsook , Arvik Torey , Chon Jung‐Whan , Nah Seung‐Yeol , Seo Kun‐ho , Bartley Glenn , Yokoyama Wallace
Published: June 2021

The objective of this study was to determine if whole grape seed could reduce plasma lipid levels and inflammatory biomarkers in adipose in obese mice. Grape seeds are high in polyphenols, including flavonols, particularly proanthocyanidins. Mice were fed either a high fat (HF, 47% fat calorie) or chow diet for 5 weeks. Mice on the HF diet that were greater in weight than mice fed chow were considered diet induced obese (DIO). The DIO mice were fed the HF or HF diet supplemented with 10% Chardonnay grape seed (CHRSD) flour for 5 weeks. Continued feeding of the HF diet supplemented with CHRSD resulted in significant decreases in weight gain, liver weight, and adipose weight despite higher feed intake. CHRSD significantly improved insulin sensitivity. These physiological improvements were accompanied by decreases in gene expression for oxidative stress, inflammation, macrophage biomarkers, and fatty acid biosynthesis in adipose tissue. Expression of genes for PPAR alpha, cholesterol synthesis, and insulin receptor were up‐regulated. These results suggest that HF‐induced obesity and insulin resistance can be ameliorated by a plant source high in flavonoids through reduction of fatty acid biosynthesis, decreased oxidative stress, and reduced inflammation in adipose tissues.
Septic Murine Pulmonary Microvascular Endothelial Barrier Dysfunction in vitro is Regulated by TIMP3
Authors: Arpino Valerie , Wang Lefeng , Bird Ryan , Pape Cynthia , Mehta Sanjay , Gill Sean
Published: June 2021

Sepsis often causes dysfunction of pulmonary microvascular endothelial cells (PMECs) leading to pulmonary edema. Metalloproteinases can regulate endothelial function through processing of cell surface proteins, including adherens junctions proteins, which has been associated with increased permeability. Tissue inhibitor of metalloproteinases 3 (TIMP3) regulates metalloproteinase function in the lung following injury. Thus, we hypothesize that TIMP3 promotes PMEC stability via inhibition of metalloproteinase activity. To test this hypothesis, we used PMECs isolated from WT and Timp3‐/‐mice. We found that TIMP3 levels (mRNA and protein) were decreased in WT PMECs in a time‐dependent fashion under septic conditions (cytomix). Analysis of leak (transendothelial electrical resistance, dextran, and albumin flux) revealed that Timp3‐/‐PMECs had significantly higher permeability under resting conditions (PBS) vs. WT PMECs. Although cytomix treatment significantly increased WT trans‐PMEC permeability, there was no effect on Timp3‐/‐PMEC barrier function. Additionally, the increased basal Timp3‐/‐ PMEC permeability was associated with disrupted surface vascular endothelial‐cadherin localization, which was rescued by treatment with GM6001, a synthetic inhibitor of metalloproteinases. Together, our data suggest that TIMP3 supports normal PMEC barrier function and that septic downregulation of TIMP3 may be an important contributor to septic PMEC barrier dysfunction.
Anti‐fibrotic Effects of Chinese Herbal Formula ALF in Mice
Authors: XIN ZHANG , Chi Man Koon , Chun Hay Ko , Tai Wai Yew , Clara Bik San Lau , Ping Chung Leung , Kwok Pui Fung
Published: June 2021

Liver fibrosis is a common pathologic outcome of all chronic liver injuries which have caused about 800,000 deaths per year worldwide which lack of effective measures to treat. This project aims to investigate the efficacy of a novel anti‐liver fibrosis herbal formula ALF, which consists of aqueous extract of Schisandrae Fructus, Artemisiae Scopariae Herba, Astragali Radix and Salviae Miltiorrhizae Radix et Rhizoma in mice. To achieve the study purpose, bile duct ligation (BDL) was used to induce liver fibrosis. 60 ICR male mice were divided into 4 groups: (1)Sham group, (2)Control group (BDL), (3)Low dose ALF group (3g/kg, BDL), (4)High dose ALF group (6g/kg, BDL). At the end of experiment, blood and liver tissues were collected. When compared with the control group, High dose ALF significantly attenuated the liver enlargement and functional injury by decreasing liver weight (p<0.05), ALT(p<0.001) and total bilirubin levels (p<0.05). Sirius Red staining also demonstrated a significant reduction in fibrosis (p<0.001). Immunohistochemical analysis showed that the deposition of collagen I and III were decreased very significantly in the ALF treatment groups (p<0.001). The fibrosis related genes MMP1a and MMP2 mRNA levels were also significantly up‐regulated, while Col1a1, Col3a1 and TIMP1 mRNA levels were significantly down‐regulated by high dose ALF(p<0.05). We have presented the first evidence that aqueous extract of herbal formula ALF showed anti‐fibrotic effects in mice.
Carnitine Palmitoyltransferase 2 (CPT2) Inhibition in Rats And Dogs Resembles Inborn Human CPT2 Deficiency
Authors: DE VERA MUDRY MARIA CRISTINA , FESTAG MATTHIAS , HOFLACK JEAN‐CHRISTOPHE , CRAMERI FLAVIO
Published: June 2021

Introduction

Carnitine palmitoyltransferases (CPT1 and 2) regulate the rate of fatty acid oxidation through intake of long‐chain fatty acids (LCFA) into mitochondria. Inhibition of CPT can reduce gluconeogenesis by decreasing LCFA and acetyl‐CoA in the liver. CPT inhibitors are therefore proposed as a potential therapeutic approach to reduce hyperglycemia in diabetes. Inborn human CPT deficiency is associated with multiorgan toxicity such as muscle weakness, rhabdomyolyis, myoglobinuria, tissue accumulation of lipids and acylcarnitines, and associated degeneration and mitochondrial damage.

Methods

To assess the safety of therapeutic CPT inhibition, a selective inhibitor of CPT2 was given daily by oral gavage to Wistar rats for 14 days, or to Beagle dogs in a single ascending‐dose phase followed by a 7‐day fixed‐dose phase.

Results

Clinical signs of CPT2 inhibition consisted of hypoactivity, salivation, emesis (dogs), and body weight loss. At high doses, rats were moribund or died after overnight fasting. Laboratory parameters, gross and histopathology, and transcriptomic profiles (rat liver) indicated liver, kidney, and muscle toxicity. Pale liver, kidney, and heart correlated with increased lipid deposits. Hepatocellular apoptosis, skeletal muscle atrophy and myodegeneration were also present. Electron microscopy of rat liver and muscle revealed mitochondrial damage.

Conclusion

On‐target toxicity of a CPT2 inhibitor in the liver, kidney, muscle, and heart in rats and dogs recapitulates and serves as a model of inherited human CPT2 deficiency.
1‐ Exposure to ambient ozone affects circulating progesterone levels and ovulation in mice.
Authors: Caruso Carla , Cabello Noe , DiAngelo Susan , Silveyra Patricia
Published: June 2021

Ambient ozone reacts with biological molecules and causes lung inflammation, oxidative stress and other health effects. Our preliminary research has shown that oxidative stress can affect the expression of fertility in rodents. Here, we investigated the effects of ozone exposure on the female reproductive axis. For this, cycling female mice were exposed to ozone (2ppm) or filter air (FA) for 3 h in the morning of proestrous (n=18) and sacrificed 4 h post‐exposure (proestrous evening). The hypothalamus, pituitary, and ovaries were harvested and pulverized, and RNA was extracted to determine gene expression of GnRH1, GnRH receptor and LH receptor. Ovarian protein expression of StAR and HSD3B1 was also detected through Western Blot. Blood was collected to determine serum pituitary and ovarian hormonal levels. A second group of animals (n=12) was sacrificed 24 h post‐exposure (morning of estrus). Ovaries were collected and histologically analyzed to determine the number of pre‐ovulatory follicules and corpora lutea. Blood was collected to compare post‐ovulatory hormone levels. We found that ozone exposure in the afternoon of proestrus decreased both hypothalamic expression of GnRH and GnRH‐receptor and circulating progesterone levels. Histological analysis in the morning of estrus revealed that ovaries from females exposed to ozone had more preovulatory follicules and less corpora lutea than females exposed to FA (p<0.05). Together, these results indicate that ozone exposure impairs ovulation in mice, potentially via affecting steroidogenesis and/or folliculogenesis.
Naltrexone‐Precipitated Opioid Withdrawal and Morphine‐Tolerance in Buprenorphine‐ or Methadone‐Treated Mice
Authors: Paronis Carol
Published: June 2021

Receptor internalization by mu‐opioid agonists has been negatively correlated to opioid tolerance and dependence. Buprenorphine does not promote mu‐opioid receptor phosphorylation yet previous reports suggest that it also does not produce opioid dependence in mice. Experiments sought to determine whether naltrexone‐precipitated withdrawal or antinociceptive‐tolerance in buprenorphine‐treated mice varies with time since the last injection; results were compared to mice treated daily with methadone (which promotes mu‐opioid receptor internalization). Naltrexone, 1‐10 mg/kg, did not elicit jumping at 4hr after 3 mg/kg/day buprenorphine yet at 24‐72hr after 0.3‐3.0 mg/kg buprenorphine, 0.1‐1.0 mg/kg naltrexone increased jumping. In contrast, the antinociceptive dose‐effect function of morphine (control ED50 = 4.2 mg/kg) was shifted 20‐fold to the right 4hr after 3 mg/kg/day buprenorphine, yet no tolerance was seen 24hr after buprenorphine. Naltrexone‐precipitated jumping also occurred at 4 and 24hr after 20 mg/kg/day methadone; in contrast to effects following buprenorphine, naltrexone was more potent at 4 than at 24hr after methadone injections. No tolerance to morphine was seen at either time point in mice treated with 20 mg/kg/day methadone, however the morphine dose‐effect function was shifted 3‐fold to the right following 45 mg/kg/day methadone. These results indicate that expression of opioid tolerance and dependence varies with time following either buprenorphine or methadone injection. (Supported by DA035411 and DA035857)
Mouse Cerebellar Neuronal Nitric Oxide Synthase (nNOS) mediates Nicotine ‐‐THC Functional Antagonism (FA) and Cross‐Tolerance
Authors: Dar M Saeed
Published: June 2021

Despite cannabinoid and nicotine co‐abuse, no study has examined co‐infused intracerebellar (ICB) THC and nicotine effect on cerebellar ataxia (CA).

Aims

(i) evaluate ICB THC + nicotine effect on CA; (ii) obtain evidence for nNOS's role in THC‐induced CA, THC – nicotine FA and cross‐tolerance.

Methods

Rotorod evaluated CA in male CD‐1 mice. All drugs [THC 20μg; nicotine 5ng; nNOS inhibitor (nNOSI; 1,2,5 ng), N’‐nitroguanidine tristrifluracetate] were given by ICB infusion.

Results

Increased THC‐induced CA by nNOSI indicated nNOS mediation. nNOSI opposed nicotine‐induced attenuation of CA by THC. The antagonism was abolished by 5 ng nNOSI. The response curves of nNOSI + nicotine + THC and aCSF + THC treatments overlapped, further evidence of nNOS mediation. We have previously observed robust cross‐tolerance between nicotine and THC. Results of ongoing chronic study involving nNOSI and nicotine infusion (once daily x 5 days followed 16 h later by acute THC) may further support nNOS role.

Discussion

Nicotine activates NOS [present in cerebellar neurons except Purkinje cells (PCs)] via release of glutamate. Alleviation of CA is likely due to: increased release of glutamate ►enhanced glutamate transmission ►diffusion of NO into PCs► activation of GC and cGMP production ►low PC firing► removal of CA. Overall, the results support a role of cerebellar nNOS in the nicotine ‐‐THC FA and cross‐tolerance.
Central Amygdala PACAP in the Behavioral Stress Response
Authors: Seiglie Mariel , Iemolo Attilio , Blasio Angelo , Cottone Pietro , Sabino Valentina
Published: June 2021

Anxiety‐related disorders are the most common forms of mental disorders; characterized by feelings of excessive worry in the absence of specific external stimuli, they are accompanied by physical, affective and behavioral symptoms.

Pituitary adenylate cyclase‐activating polypeptide (PACAP) is a 38‐amino acid peptide located in the hypothalamus and in various extra‐hypothalamic regions including the central nucleus of the amygdala (CeA). PACAP and PAC1 receptor (PAC1R) have been proposed to play a key role in mediating the behavioral and endocrine responses to stress; however, few studies have examined the involvement of the extrahypothalamic PACAP system in the modulation of stress. Our aim was to elucidate the role of the PACAP/PAC1 system of the CeA in the context of anxiety.

We first assessed the effects of PACAP microinfused into the CeA of male rats in the elevated plus maze (EPM) and the acoustic startle response (ASR), behavioral tests sensitive to states of anxiety and fear. PACAP was also given into the basolateral amygdala (BlA) as a control brain region. We then used restraint stress to determine whether the endogenous PACAP‐PAC1 system is recruited by acute stressors.

Infusion of PACAP into the CeA significantly reduced percent open arm time in the EPM and increased plasma corticosterone levels, while BlA microinfusion had no effect. Infusion of PACAP into the CeA also increased acoustic startle reactivity. Finally, we observed that an acute, 1h exposure to restraint produced a rapid elevation of PACAP and PAC1R expression in the CeA, but not the BlA.

These data prove an anxiogenic role for the PACAP/PAC1 system of the CeA and suggest that hyperactivity of this system may underlie the anxiogenic effects of stress.
Pharmacological treatment(?) of Sleep Problems in a Swedish Pediatric Setting
Authors: Holmbäck Ulf , Forslund Anders
Published: June 2021

Aim

The purpose of this study is to investigate the clinical use of sleep aids, specifically melatonin for sleep disorders in children and adolescents in treated at various facilities in Uppsala County, Sweden

Method

A cross sectional study was conducted using a short 9 question web‐based survey. The survey contained questions about sleep medications in general and some specific questions about melatonin/Circadin®. The questionnaire was then sent to doctors in various clinics in Uppsala County, Sweden.

Results

Preliminary results shows that the majority of the doctors prescribe some sort of sleep aid, where Circadin® was the most commonly prescribed sleep aid; with melatonin as a close second (Melatonin not an OTC‐drug in Sweden). Sleep aid was prescribed to children of all ages; and most often in conjunction to various neuropsychiatric disorders. The average dose was 2‐3 mg melatonin /Circadin® per day, with some prescribing up to 6 or 9 mg per day.

Conclusion

These preliminary results indicate that the doctors prescribe Circadin® to a larger degree. This despite that Circadin®'s current indication is for people >55 years old. Moreover, high doses of melatonin are prescribed, most probably leading to next‐day drowsiness and sub‐optimal functioning of the children. Doctors need to be informed about proper choice of drug and dose for the pediatric patient
Nicotine Increases Codeine Analgesia Through the Induction of Brain CYP2D Central Activation of Codeine to Morphine
Authors: McMillan Douglas , Tyndale Rachel
Published: June 2021

Cytochrome P450 2D (CYP2D) metabolizes codeine to morphine, a step required for codeine analgesia. However lower morphine permeability across the blood‐brain barrier, as well as exportation of morphine, suggest that after codeine the initial morphine in the brain, and resulting analgesia, may be due to brain CYP2D activation. Higher brain, but not liver, CYP2D6 is found in smokers, and 7‐day nicotine treatment induces rat brain, but not liver, CYP2D. We hypothesized that inducing rat‐brain CYP2D would increase the CNS CYP2D‐mediated activation of codeine to morphine resulting in increased analgesia, despite first‐pass metabolism, which could be blocked by inhibition of brain CYP2D. Codeine (20mg/kg ip) analgesia was tested: compared to pretreatment with vehicle, 7‐day nicotine (1mg/kg sc, brain CYP2D inducer) increased analgesia (1.59, AUC0‐30min fold change vs. vehicle; p<0.001), propranolol (20 μg in 4 μl icv, brain CYP2D inhibitor) decreased analgesia (0.56; p<0.05), and co‐pretreatment was similar to vehicle (1.23; p>0.1). 7‐day nicotine increased and propranolol decreased brain, but not plasma, morphine levels; analgesia correlated with brain (p<0.02), but not plasma (p>0.4), morphine levels at 15 min after codeine. Pretreatments (alone or together) had no effect on baseline nociception or morphine (3.5mg/kg ip) analgesia. Thus, variation in brain CYP2D activity may contribute to clinically relevant differences in individual response to centrally acting CYP2D substrates. Individuals exposed to nicotine may have increased brain metabolism contributing to altered drug efficacy and abuse liability.

Funding: CIHR and the CAMH Foundation
Combined Treatment with Morphine and Δ9‐Tetrahydrocannibinol (THC) in Rhesus Monkeys: Antinociceptive Tolerance and Withdrawal
Authors: Gerak Lisa , France Charles
Published: June 2021

Opioid receptor agonists are effective for treating pain; however, tolerance and dependence can develop with repeated treatment. Combining opioids with cannabinoids can enhance their analgesic potency, although it is less clear whether combined treatment alters opioid tolerance and dependence. In this study, 4 monkeys received 3.2 mg/kg morphine alone or in combination with 1 mg/kg THC; the antinociceptive effects (warm water tail withdrawal) of morphine and the cannabinoid receptor agonists WIN 55212 and CP 55940 were determined before and during treatment. To determine whether this treatment produced dependence, behavioral signs indicative of withdrawal were monitored when treatment was discontinued. Before treatment, morphine and the cannabinoids, given alone, increased tail withdrawal latency to 20 sec (maximum possible effect). During treatment, latencies did not exceed 10 sec for any drug, up to doses 3‐ to 10‐fold larger than those that were fully effective before treatment. Rightward and downward shifts in antinociceptive dose‐effect curves were greater in monkeys receiving the morphine/THC combination. Withdrawal signs included increased heart rate, decreased activity, and increased tongue movement, and these signs were similar in monkeys that received morphine alone or in combination with THC. These results demonstrate that antinociceptive tolerance is greater during treatment with the combination, whereas dependence and withdrawal are not changed. When combined with THC, a much smaller dose of morphine is needed to produce antinociceptive effects, and the ability of THC to enhance tolerance to small doses of morphine will be examined in the next study. Supported by R01DA005018 and K05DA17918.
Effect of co‐ultramicronized Palmitoylethanolamide and Luteolin Association on Neurogenesis in a Mouse Model of Spinal Cord Injury
Authors: Cordaro Marika , Paterniti Irene , Siracusa Rosalba , Impellizzeri Daniela , Crupi Rosalia , Bruschetta Giuseppe , Esposito Emanuela , Cuzzocrea Salvatore
Published: June 2021

Neurogenesis has been described in various regions of the CNS throughout life. Little is known, however, regarding the nature or extent of new cell division after spinal cord injury (SCI) in mice and about its potential to contribute to endogenous repair mechanisms. In a recent study, we have demonstrated that treatment with a new compound, a formulation including palmitoylethanolamide and luteolin, co‐ultraPEALut, significantly reduced inflammatory secondary damage associated with SCI. Thus, the aim of this study was to investigate the neuroprotective effect of co‐ultraPEALut in the injury‐stimulate neurogenesis in a mouse model of SCI. SCI was induced in mice by the application of vascular clips to the dura via a four‐level T5 to T8 laminectomy. The results of the present study demonstrated that the administration of co‐ultraPEALut increased bromodeoxyuridine and doublecortin immunoreactive cells in the spinal cord frommice subjected to SCI. This neuronal development correlated with synaptic plasticity, identified using the Golgi impregnation method to quantify dendritic spines in spinal cord. In addition, co‐ultraPEALut treatment also increased the expression of several neurotrophic factors such as BDNF, GDNF, NGF and NT3. The present study demonstrated that co‐ultraPEALut could have a role on birth, survival, and differentiation of new neurons and maturation of spines in the spinal cord and could represent could represent a promising strategy for traumatic diseases.
Fumaric Acid Esters Attenuate Secondary Degeneration and Promote Functional Recovery Following Experimental Spinal Cord Injury
Authors: Esposito Emanuela , Paterniti Irene , Cordaro Marika , Cuzzocrea Salvatore
Published: June 2021

Spinal cord injury (SCI) causes a significant physical, emotional, social and economic burden. The main clinical hallmark of SCI is the permanent loss of motor, sensory and autonomic functions below the level of injury that are in part due to the ensuing inflammatory response. Fumaric acid esters (FAEs) display immunomodulatory properties and may also exert neuroprotective effects, as shown in vitro as well as in experimental models of multiple sclerosis.

The aim of the present study was to evaluate the potential beneficial effects of FAE in a mouse model of traumatic spinal cord injury. SCI was induced in mice by spinal cord compression, and dimethyl fumarate (DMF) and monomethyl fumarate (MF) (both at 30 mg/kg) were orally administered to the mice 1 and 6 h after SCI, and once daily thereafter for 10 d. Motor function (Basso Mouse Scale) improved gradually in mice treated with DMF. However, those treated with DMF exhibited a significantly more rapid and sustained recovery of motor function. FAEs also significantly reduced the severity of inflammation (proinflammatory cytokines, apoptosis, and neuroptrophic factors) that characterized the secondary effects of SCI. Again, the effects of DMF were superior to those of MF for several parameters. These results showed marked protective effects of DMF in an animal model of SCI, significantly enhancing recovery of motor function, possibly by reducing the secondary inflammation and tissue injury that characterize this model. In summary, DMF may constitute a promising target for future SCI therapies.
Spinal Cord Injury in Rat – Therapeutic Effect of Pentadecapeptide BPC 157
Authors: Perovic Darko , Buljat Gojko , Kolenc Danijela , Drmic Domagoj , Seiwerth Sven , Sikiric Predrag
Published: June 2021

Background

Previously pentadecapeptide BPC 157 exhibited a strong healing effect on brain traumas and peripheral nerve injuries (Regul Pept. 2010;160(1‐3):26‐32; Regul Pept. 2010;160(1‐3):33‐41). Aim. To verify influence of BPC 157 on rat spinal cord injury. Materials and methods. Deeply anaesthetized Wistar Albino male rats, 350 g b.w., were subjected to surgery (all experimental procedures were approved by the local Ethics Committee): after laminectomy on level L2‐L3 (correspond to sacrocaudal spinal cord) 60 second compression with neurosurgical piston (60‐66 g) on exposed dural sac. One single medication (IP) (saline 5ml/kg; pentadekapeptide BPC 157 200µg/kg; 2 µg/kg) was 10 min post‐injury and sacrifice was at 1, 2, 4, 12, 26 and 52 weeks. Tail motor function (1 paralysis; 5 normal) and spasticity (1 normal; 5 maximum) was scored daily. Histology samples of spinal cord were taken from lesion site, and tail samples 10 mm distal from base. Results. Tail motor function. BPC 157 rats have significantly higher motor score in all investigated time. Tail spasticity. BPC 157 rats have significantly less spasticity starting from 4 weeks to the end of experiments. Histology of spinal cord. From 2 weeks to the end of follow up BPC 157 rats have less grey matter edema and less cyst and axonal necrosis in white matter. Histology of rat tail. All treatment groups show increased number of all and small axons starting from 4 weeks but without statistically difference between groups. Conclusions. Given after sacrocaudal spinal cord injury, BPC 157 recovered rat tail function with single treatment. Specifically, BPC 157 decreased secondary injury of spinal cord, and consequently, improved neurologic function recovery. Supported by Grant 108‐1083570‐3635, Croatia.
Rgs8‐16 are Required for Energy Homeostasis in Pancreatic Cancer‐Associated Malnutrition
Authors: Zolghadri Yalda , Ocal Ozhan , Wilkie Thomas
Published: June 2021

Pancreatic insufficiency develops by about 5 weeks of age in KC (LSL‐KrasG12D;p48Cre) mice that express KrasG12D in all pancreas cells. Kras mutations (e.g. KrasG12D) are found in over 90% of human PDA and are an early event in a multistep process leading to PDA. KrasG12D causes dedifferentiation of acinar cells and a drastic reduction in digestive enzymes secreted by the pancreas. KC mice become malnourished but can survive over one year before succumbing to Pancreatic Ductal Adenocarcinoma (PDA). Protein kinase and G‐Protein Coupled Receptor (GPCR) signaling can initiate Ras activation. Regulators of G‐protein Signaling (RGS) proteins are GTPase Activating Proteins (GAPs) that regulate the intensity and duration of Gi‐ and Gq‐coupled GPCR signaling. Activating mutations in Gq have been associated with PDA. We find Rgs8 and Rgs16 expression is induced in Pancreatic Intraepithelial Neoplasia (PanIN) and Intraductal Papillary Mucinous Neoplasm (IPMN) in KC mice by 2 weeks of age. We crossed the Rgs8‐16 double knockout into KC (termed KC‐R) mice to test if Rgs8‐16 are tumor suppressor genes. Most KC‐R mice die before 4 months of age because they can not maintain energy homeostasis to counteract pancreatic insufficiency – Rgs8‐16 are required in liver to conserve energy utilization during malnutrition. Consistent with this finding, the effects of caloric restriction are more severe in Rgs8‐16 dko compared to wild type mice. The effects of Rgs8‐16 deficiency on exocrine pancreas function, acinar‐to‐ductal metaplasia, apoptosis and tumor progression in KC‐R mice are under investigation. Supported by NCI CA161624.