bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of fiber on meal patterns and weight loss in the rat
Authors: Thaw Andrew Kurt , Brooks Rachel , Wootten Abbott , Mong Brandon
Published: June 2021

Satiety and weight loss enhancement due to added dietary fiber has received considerable attention recently. Most studies have examined the effects of fiber on a single test meal, however the temporal relationship of fiber and reduced hunger has not been well stated. For example, does fiber affect meal size or intermeal interval; does it last for several hours or acutely? Here we use a 24 hour food monitoring system to examine the discreet eating patterns of rats following a pre‐meal snack of low, medium or high dose of fiber in a sucrose solution. 24 male Sprague‐Dawley rats (age = 75 days) were maintained on standard chow and distilled water. Following 2 weeks of acclimation subjects received a .25M sucrose solution mixed with 1, 3 or 6 grams/liter of fiber (2 hour access). Water and food were removed during the fiber presentation and returned immediately after. Measures of daily food intake, weight gain, meal size, intermeal interval, etc. were analyzed to determine the discrete and chronic effects of fiber on meal patterns. Results indicate a significant acute satiety effect followed by normal eating patterns within 2 hours post‐fiber. There were no significant weight differences in control or experimental rats for any of the fiber concentrations. We conclude that fiber may be useful in caloric reduction for single meals, but provide minimal weight loss benefits.
Histochemical Imaging of Brain Circuits that Regulate Food Intake
Authors: Blevins James Ernest
Published: June 2021

We hypothesize that food intake during meals is regulated by a population of hypothalamic paraventricular (PVN) oxytocin (OXY) neurons that have direct projections to the nucleus tractus solitarius (NTS) in the hindbrain and enhance the behavioral and neuronal responses to cholecystokinin (CCK‐8), a satiety signal from the intestines. PVN OXY neurons terminate among NTS neurons that show Fos activation in response to CCK‐8. To identify OXY‐sensitive NTS neurons, we used laser capture microdissection (LCM) with RT‐PCR to demonstrate a reduction of neuropeptide Y (NPY) and OXY receptor (OXYr) mRNA in the NTS following injections of the neurotoxin OXY‐saporin. Retrograde tracer cholera toxin B injections into the NTS labeled OXY cells in the PVN. Fos immunoreactivity was increased in OXY PVN neurons following treatment with the adipocyte hormone leptin, which potentiates the satiety actions of CCK‐8 via CNS melanocortin‐4 receptors (MC4r). Using LCM and RT‐PCR, we identified MC4r mRNA in PVN neurons that project to the NTS. These findings indicate that leptin's anorectic actions in the CNS are mediated by MC4r signaling in PVN OXY neurons that project to the NTS in the hindbrain. Support: Dept. Veterans Affairs, NIH DK17047, DK52989, 5P30 DK35816‐18, NS32273, Advanced Targeting Systems.
Body self‐esteem and formula use in the first 24 hours postpartum among low‐income, obese Latinas
Authors: Chapman Donna J , Wetzel Katherine , Bermudez‐Millan Angela , Damio Grace , Kyer Nan , Young Sara , Pérez‐Escamilla Rafael
Published: June 2021

The objective of this study is to identify risk factors for partial (vs exclusive) breastfeeding (BF) in a sample of low‐income, obese Latinas. We analyzed results from an ongoing randomized trial evaluating the efficacy of a specialized BF peer counseling intervention targeting obese women. Data were obtained prenatally and on day 1 postpartum (pp), including infant feeding practices, body esteem ratings, demographics and medical record review. Bivariate analyses were conducted to identify variables associated with the use of infant formula in the first 24 hours pp, with significant variables entered into a logistic regression model. Among those with exclusive breastfeeding (EBF) data on day 1 (n=69), 98.6% initiated BF and 58.0% had introduced non‐breastmilk fluids. Formula was used by 56.5% and formula, water and sugar water were used by 1.5%. In bivariate analyses, women with a high (vs low) body esteem score for weight‐sensitive body parts were less likely to EBF (p=0.025) and Puerto Rican women (vs all others) tended to be less likely to EBF (p = 0.058). In multivariate analyses, the only significant risk factor for partial BF by 24 h pp was a high body esteem score (OR = 2.98 (95% CI: 1.08, 8.26). These preliminary findings suggest that in this sample of obese women, those who are more satisfied with their body are less likely to EBF on day 1 pp. Funded by the Donaghue Foundation and NIH/NCMHD grant #P20MD001765.
Urine volume increases and urine specific gravity decreases in cats with nephrolithiasis or urolithiasis
Authors: Yu Shiguang , Burris Patricia
Published: June 2021

The etiology of urinary tract stones in cats is not clear. Among others, concentrated urine is considered a risk factor. The objective of this study was to compare urine measurements in stone‐forming cats with those in healthy cats.

Four cats with kidney (3 cats) or urinary bladder (1 cat) stones and 11 healthy cats were used. Stone‐forming cats were 6 to15 years old (median = 12.7 years) and healthy cats were 5 to 13 years old (median = 10.8 years). The stone‐forming cats were confirmed with x‐ray radiography and/or ultrasonography.

Cats were fed a commercial dry cat food for 4 weeks. Forty‐eight hour urine was collected during the last two days of the study. The urine was collected continuously in the collection receptacles kept at 38 °C.

Comparing with healthy cats, stone‐forming cats had significantly increased urine volume (87 ml/day vs. 145 ml/day, p < 0.05) and decreased urine specific gravity (1.033 vs. 1.025, p < 0.05). Urine pH was similar between the groups (5.94 vs. 5.88, p = 0.604). Urinary NH4+, Na, K, P, and Oxalate were significantly lower in stone‐forming cats (p < 0.05). Urinary Cl, Ca, Mg, S, and Citrate were similar between the groups (p > 0.05). These data show that urine was not concentrated in cats with existing stones in the urinary tract. The mechanism underling the increased urine volume and decreased urine specific gravity in stone‐forming cats is unclear and needs further investigation.
Menopausal status modified the association between body mass index and hypertension among Filipino women
Authors: Agustin Sonny Sombra , Adair Linda , Gultiano Socorro , Bas Abet , Lee Nanette
Published: June 2021

The study aimed to determine whether the association between body mass index (BMI) and hypertension is modified by menopausal status among Filipino women. Participants included 1,804 women (aged 38 to 71 yr) from the Cebu Longitudinal Health and Nutrition Survey. Hypertension was defined as systolic blood pressure >= 140 mmHg or diastolic blood pressure >=90 mmHg, BMI was calculated using measured weight and height, and menopausal status was based on self reports over multiple survey years. Controlling for age, smoking and fasting blood glucose levels, logistic regression analysis showed that BMI increased hypertension odds in all women. Using likelihood ratio test, we found significant interaction between BMI and menopausal status (p<0.05). Compared to premenopausal women, the effect of BMI on hypertension odds was lower among post‐menopausal women (odds ratio (95% confidence interval): 1.14, (1.10, 1.18) versus 1.09 (1.05, 1.12), respectively). Results suggest that BMI may be more of a risk factor before menopause possibly due to different competing risks associated with menopausal status. NIH 1 R03TW008133‐01, 1 R01 HL085144
Influence of Menopause on the Risk of Cardiovascular Disease and Bone Loss in the Postmenopausal Women
Authors: Malepati Dhananjaya Naidu , Goudu Sivanarayana , G Jagadesh
Published: June 2021

Menopause is considered to be a cardiovascular risk factor and exogenous hormone therapy after menopause was demonstrated to contribute to heart health. In the present study, the effect of menopause on cardiovascular disease (CVD) and bone resorption has been investigated. The incidence of coronary heart disease and the risk of osteoporosis increase sharply after menopause in women. We compared 96 postmenopausal women (study group) and 40 premenopausal women (control group). Their postmenopausal status was confirmed by serum estradiol levels. Compared to premenopausal women, postmenopausal women had significantly increased the levels of total cholesterol (TC) (P<0.001), atherogenic index (TC/high density lipoprotein) (P<0.001), urine C‐telopeptides of type I collagen (CTX‐I) (P<0.001) and serum tartrate resistant Acid Phosphatase‐5 b (TRACP‐5B) (P < 0.001). These results suggest that the menopausal status may have a significant effect on the primary cardiovascular risk factors such as total cholesterol, atherogenic index and the risk of osteoporosis (TRACP‐5B, CTX‐I) in the postmenopausasl women. This study suggests that the Physicians should consider both the risk of CVD and bone loss while initiating the therapies.
Nutritional status assessed by patient‐generated subjective global assessment is associated with diet quality and quality of life in Korean cerebral infarction patients
Authors: Lim Hyunjung , Park Jin Hee , Kim Jin Kyung , Choue Ryowon
Published: June 2021

Assessment of the nutritional status of cerebral infarction (CI) patients is important because their nutritional status influences disease outcome. The aim of this study was to assess the nutritional status of CI patients using the scored patient‐generated subjective global assessment (PG‐SGA) and to investigate the relationship between the quality of diet and life based on their nutritional status. 73 CI patients were recruited from Kyung Hee University Oriental Medical Center in Seoul, Korea from May to July 2007. The subjects' PG‐SGA, dietary diversity score (DDS), dietary variety score (DVS), diet quality index‐international (DQI‐I), and stroke‐specific quality of life (SSQOL) were investigated. The patients were classified by PG‐SGA categories as well‐nourished (26.0%), moderately malnourished (49.3%), or severely malnourished (24.7%). The DDS and DVS were significantly lower in severely malnourished patients. The DQI‐I was significantly greater in the well‐nourished group. The SS‐QOL was significantly lower in the malnourished group. Quality of diet and life were significantly correlated with their nutritional status. In conclusion, PG‐SGA is a useful nutritional assessment tool for CI patients with stable vital signs. The well‐nourished patients demonstrated better diet quality, as well as better quality of life.
Increased adiponectin among Cuban American type 2 diabetics with metabolic syndrome
Authors: Knight Sepulveda Karina , Huffman Fatma G. , Mclean Michael , Zarini Gustavo G.
Published: June 2021

Metabolic syndrome (MetS), defined by clustering of multiple CVD risk factors, is associated with development of type 2 diabetes (T2D) and CVD. Circulating adiponectin decreases with obesity, MetS, T2D and CVD. Subjects were randomly recruited from Miami‐Dade and Broward Counties, FL. MetS was determined using the ATP III criteria. Fasted whole blood for adiponectin was collected into tubes containing K3EDTA. Adiponectin was measured using ELISA (Linco Research Inc). Descriptive statistics, Pearson's correlation, and logistic regression model were used for data analysis. The model included: main effect, main effects + interaction, and main effects, interaction, controlling for age and gender. Individuals were: 30% with MetS, 59% female, 69% non‐diabetic, 61.06±11 years old, had BMI 30.24±5.38, mean log adiponectin of 3.02±1.29. There was an inverse correlation (r=‐0.453, P<.01) between log adiponectin and diabetes status. Logistic regression analyses of the presence or absence of MetS among diabetics and non‐diabetics were significant for diabetes status (P<.001), interaction for log adiponectin by diabetes status (P<.002) and BMI (P<.001), controlled for age and gender. In conclusion adiponectin was higher in T2D and MetS subjects.
Comparison of nutrient intakes from a detailed inventory to a supplement frequency questionnaire: results from the SURE Study
Authors: Monroe Kristine R , Murphy Suzanne , Wilkens Lynne , Steffen Alana , Albright Cheryl , Yonemori Kim , Morimoto Yukiko
Published: June 2021

The Supplement Reporting (SURE) Study evaluated the most common methods of collecting dietary supplement use data among five racial/ethnic populations participating in the Multiethnic Cohort Study in Hawaii and Los Angeles. Here we compare data from a detailed, full year inventory of supplement use to estimates from a self‐administered supplement frequency questionnaire (SFQ) that asked about multivitamin use, 11 single‐nutrient supplements, as well as fish oil and garlic. Categorical dose range was asked for vitamins C and E only. We present here the overall results for four common components of supplements for 399 subjects who completed the 1‐year study. The percentage of individuals where the difference in the nutrient values between the inventory and the SFQ were larger in absolute value than 20% of the mean inventory value, using logged values, were 21% for folate, 26% for calcium, 33% for vitamin C and 47% for vitamin E. Weighted kappa statistics were 0.48 for folate, 0.50 for vitamin E, 0.58 for vitamin C and 0.59 for calcium. For overall nutrient intakes, providing information on dosage does not appear to have increased the accuracy of the estimates of vitamin C and vitamin E intakes relative to the other two nutrients. We conclude that the SFQ might be used successfully when ranking is important, but may not be the optimal instrument when absolute nutrient intake is required. Supported by NCI grant R01 CA106744.
Association of age, socioeconomic status (SES) and acculturation with intake of omega‐3 fatty acids in Latinas
Authors: Lora Karina Regina , Lewis Nancy , Eskridge Kent , Travnicek Daryl
Published: June 2021

We examined the association of age, SES (education, income, employment) and acculturation on total omega‐3 fatty acid (T n‐3), ALA and LC n‐3PUFA (EPA, DHA) food intakes of Midwestern Latinas. Participants (n=162) were first generation Latinas, 20 to 50 y, non‐pregnant and non‐lactating at the time of the study. Sociodemographic, dietary intake by use of a validated n‐3 food frequency questionnaire and acculturation information were collected. Intakes of T n‐3, ALA and LC n‐3 PUFA (g) were 1.2±0.7, 1.1±0.6, and 0.1±0.1 respectively. Eighty‐three percent of participants were low acculturation. Age was negatively correlated with T n‐3 and ALA intakes. Education, income and acculturation were positively correlated with LC n‐3 PUFA intakes (P<0.05). Latinas in the 20‐30 y age category compared to the 41‐50 y age category consumed 1.2 and 1.3 more times of T n‐3 and ALA respectively. Latinas who completed =4 y of college compared to those with =12 y of education, had incomes =$50,000 compared to those with incomes of <$10,000, and were high acculturation consumed 1.5, 1.7 and 1.5 more times LC n‐3 PUFA respectively. Age was related to T n‐3 and ALA intakes after accounting for variation in education, income and acculturation; however, only income was related to LC n‐3 PUFA intake when the other variables were taken into account (P<0.05). Latinas met the Adequate Intake for ALA, but had intakes < recommended 500 mg/d of EPA and DHA.
The Liver X‐Receptor (LXR) gene promoter is hypermethylated in a mouse model of prenatal protein restriction
Authors: Plosch Torsten , van Straten Esther M.E. , Bloks Vincent W. , Huijkman Nicolette C.A. , Kuipers Folkert
Published: June 2021

Prenatal nutrition, e.g., nutritional status of the mother, has been epidemiologically identified as a determinant of adult disease. Feeding low‐protein diets during pregnancy in rodents is a well‐established model to induce "programming" events in offspring. We hypothesized that protein restriction would induce epigenetic adaptations which would interfere with lipid metabolism. C57BL/6 mice were fed a protein restricted diet during pregnancy. At day 19.5 of pregnancy, dams and fetuses were sacrificed. CpG island methylation microarrays were performed on fetal liver DNA. 204 promoter regions were differentially methylated upon protein restriction. The liver X‐receptor (Lxr) alpha promoter was hypermethylated in protein‐restricted pups. Lxr alpha is a nuclear receptor critically involved in control of lipid metabolism. mRNA level of Lxra tended to be reduced in fetal liver upon maternal protein restriction, whereas expression of Lxr target genes were significantly reduced. In vitro methylation of a mouse Lxra‐promoter/luciferase expression cassette resulted in a 24‐fold transcriptional repression. Our study demonstrates that, in mice, protein restriction during pregnancy interferes with DNA methylation in fetal liver. Lxra is a target of differential methylation and Lxra transcription is dependent on DNA methylation. We suggest that prenatal nutrition may influence adult lipid metabolism by DNA methylation which may contribute to the epidemiological relation between prenatal/neonatal nutrition and adult disease.

Supported by the Dutch Heart Foundation (2004T048).
MTHFR C677T genotype and choline intake affect choline dynamics in Mexican American men
Authors: Caudill Marie A , Yan Jian , Wang Wei , Gregory Jesse F
Published: June 2021

As part of a 12 wk controlled feeding study conducted in Mexican American men, stable isotope methodology was employed to assess the influence of choline intake and MTHFR C677T genotype on choline flux. Beginning wk 10, Mexican American men (15 677TT, 8 677CC) consumed 15% of either 550 or 1100 mg/d choline as D9‐choline. Liquid chromatography (or gas chromatography) mass spectrometry was used to measure the isotopic enrichment of choline and relevant metabolites in urine and blood (on‐going) obtained at wk 12. In urine, the enrichment of D6‐dimethylglycine to D9‐choline was greater (P<0.05) in men with the MTHFR 677TT vs CC genotype. In addition, enrichment of betaine was greater (P<0.05) in the 1100 vs the 550 mg/d choline intake group. These initial results suggest that choline flux through the betaine N‐methyltransferase pathway is greater in men with the MTHFR 677TT genotype relative to those with 677CC genotype under the conditions of this study. Further, oxidation of choline to betaine appears to be enhanced when choline intake is increased.

Supported by the National Institutes of Health grant S06GM53933 and funds from the California Agricultural Research Initiative.
Characterization of homocysteine and methyl group metabolism in female non‐obese diabetic (NOD) mice
Authors: Nieman Kristin M , McGuire Annette S , Schalinske Kevin L
Published: June 2021

Methyl groups are critical for numerous biological reactions and proper regulation of methyl group and homocysteine (hcy) metabolism is essential for disease prevention. The hepatic protein glycine N‐methyltransferase (GNMT) is a key regulator of supply and utilization of methyl groups from the primary donor, S‐adenosylmethionine. Chemically‐induced type 1 diabetes has been shown to alter hcy and methyl group metabolism, resulting in a methyl deficient state, including DNA hypomethylation. The objective of this study was to characterize hcy and methyl group metabolism using a genetic model of diabetes, the non‐obese diabetic (NOD) mouse, which spontaneously develops autoimmune‐mediated type 1 diabetes. NOD mice and their controls (ICR), were sacrificed at 12, 18, and 24 wk of age for collection of blood and liver tissue. Plasma hcy concentrations in NOD mice were only 63% of the control values in 18 wk old ICR animals. NOD mice exhibited a 1.4‐fold increase in GNMT activity compared to ICR mice at 18 wk. Neither metabolic parameter was altered in 12 wk old animals. The NOD mouse, a genetic model for type 1 diabetes, exhibits similar perturbations in methyl group and hcy metabolism as the chemically‐induced rat model. This validation will be important in future studies directed at understanding the relation between diabetes, perturbation of methyl groups, and specific pathologies (Support: NIH HD057315‐02).
Inhibition of cigarette smoke‐induced lung cell proliferation by dietary selenium in mice
Authors: Li Jun , Tharappel Job C. , Han Sung Gu , Cantor Austin H. , Lee Eun Y. , Gairola C. Gary , Glauert Howard P.
Published: June 2021

The objective of this study was to determine if dietary selenium could inhibit the induction of pulmonary cell proliferation by cigarette smoke in mice. Female A/J mice were fed purified diets containing 0.15, 0.5, or 2.0 mg/kg selenium in the form of sodium selenite. After 3 weeks, mice in each diet group were divided into two subgroups; one used as control while the other was exposed to cigarette smoke for five consecutive days and euthanized 3 days later. During this time mice were administered bromodeoxyuridine (BrdU) in the drinking water. After euthanasia, the left lung lobe was processed for serial sectioning and cell proliferation analysis. Cigarette smoke increased cell proliferation in the terminal bronchioles and large airways, but not in alveoli. Selenium inhibited cell proliferation in the terminal bronchioles and large airways areas. Increasing the dietary selenium level led to the increased selenium levels in the blood and lung, and increased glutathione peroxidase activity in the serum. Cytochrome P‐450 1A1 protein levels in the lung were increased by cigarette smoke but were not affected by dietary selenium. It is concluded that dietary selenium inhibits pulmonary cell proliferation caused by cigarette smoke exposure and that this inhibition may be related to selenium levels in the lung. (Supported by NIH CA125788, the China Scholarship Council, and the Kentucky Agricultural Experiment Station)
Effect of non‐oil seed pulses on glycemic control: A meta‐analysis of randomized controlled trials in humans.
Authors: Sievenpiper John , Esfahani Amin , Wong Julia , Carleton Amanda , Jiang Henry , Bazinet Richard , Jenkins David , Kendall Cyril
Published: June 2021

Background

Dietary pulses (beans, lentils, chickpeas, etc.) are a good source of viscous fibre and a valuable means for lowering the glycemic‐index (GI) of the diet.

Objective

To assess the effect of pulses on glycemic control, we conducted a meta‐analysis of experimental trials investigating the effect of pulses alone or in low‐GI or high‐fibre diets on indices of glycemic control.

Methods

We searched MEDLINE, EMBASE, CINAHL, and Cochrane Library for relevant controlled trials of ≥7d in humans. Two independent reviewers extracted information on study design, participants, treatments, and outcomes. Generic inverse variance models were used for pooled analyses. Heterogeneity was assessed by Chi2 and quantified by I2.

Results

Forty‐four trials were included. Pulses alone (12 trials) lowered fasting blood glucose (FBG) (standardized mean difference [SMD] ‐0.81 [95% CI ‐1.32,‐0.29]) and insulin (‐0.61[‐1.07,‐0.14]). Pulses in low‐GI‐diets (19 trials) lowered glycosylated proteins (GPs) (‐0.28[‐0.42, ‐0.14]). Pulses in high‐fibre diets (12 trials) lowered FBG (‐0.22[‐0.43,‐0.00]) and GPs (‐0.33[‐0.59,‐0.06]). Inter‐study heterogeneity was significant in all cases.

Conclusions

These data demonstrate that pulses alone or in low‐GI or high‐fibre diets improve markers of glycemic control in humans. Subgroup analyses are planned to explore sources of heterogeneity.
Effects of a leucine‐containing nutraceutical on fat oxidation in overweight and obese adults
Authors: Zemel Michael B , Sun Xiaocun
Published: June 2021

We have previously shown leucine to modulate energy partitioning between adipose tissue and myocytes by suppressing adipocyte lipid storage, increasing myocyte fatty acid oxidation and stimulating mitochondrial biogenesis in both cell types. Consequently, we have now evaluated the efficacy of a nutraceutical preparation (InnutriaTM) containing 2.25 g leucine/d in 3 daily doses on fat oxidation. 20 overweight and obese subjects were randomized to receive Innutria‐supplemented or control tea three times/day for 28 days. All subjects maintained their usual diets and modestly increased their physical activity by walking (steps increased from baseline 3882+713 steps/day to 8406+925 steps/day during treatment; no difference between groups). Respiratory quotient (RQ) and fat oxidation were assessed by respiratory gas exchange on day 0 and day 28. There were no significant changes in RQ or fat oxidation in the control group (RQ change, 0.011; fat oxidation change, ‐0.23+0.28 g/hr, ‐5.6 g/d, NS), while the supplemented group exhibited a decrease in RQ (‐0.019, p<0.04) and a corresponding increase in fat oxidation (+1.4+0.4 g/hr, 33.6 g/d, p<0.01). These data demonstrate that a nutraceutical blend containing leucine effectively increases fat oxidation and may thereby contribute to weight management in overweight and obese subjects.
Impact of Contaminant Heterogeneity on Anticoagulation and Adverse Reactions Associated with Recalled Heparin
Authors: Fareed Jawed , Hoppensteadt Debra , Jeske Walter , Adiguzal Cafer , Iqbal Omer , Walenga Jeanine M.
Published: June 2021

It has been assumed that different batches of contaminated heparin contained similar forms of oversulfated chondroitin sulfate (OSCS). Contaminants were isolated from four batches of UFH and two batches of LMWH by digestion of heparin followed by alcohol precipitation and ion‐exchange chromatography. Biologic activity was measured in terms of anticoagulant activity, thrombin generation inhibition, protamine/PF4 neutralization, and interaction with AT and HCII. The contaminated UFHs had different anti‐Xa:anti‐IIa ratios (0.93‐1.24) and amount of heparinase resistant material (14‐30%) and two contained significant amounts of dermatan sulfate. Isolated contaminants exhibited distinct PF4 and protamine neutralization profiles. LMWHs had comparable molecular weights and biologic actions, but differed in heparinase‐1 digestion profile. The molecular weight of the LMWH contaminant was lower (12.8 vs. 14.1‐16.8 kDa) and exhibited differences in thrombin generation inhibition. The contaminants isolated from heparin and LMWH had potencies of 28‐46 and 38‐46 USP U/mg, respectively. Contaminants isolated from recalled batches of heparin are heterogenous. Moreover, the contaminants obtained form LMWHs may exhibit additional structural and biologic differences. The variations observed in the adverse reactions with recalled heparins may be due to compositional variations in the contaminants.
mTOR‐dependent IL‐10 expression inhibits LPS induction of tissue factor and cytokines in macrophages.
Authors: Baker Alyson K , Wang Ruipeng , Mackman Nigel , Luyendyk James P
Published: June 2021

Bacterial lipopolysaccharide (LPS) induces monocytes/macrophages to express proinflammatory cytokines and tissue factor (TF), the primary activator of the coagulation cascade. Anti‐inflammatory signaling pathways including the phosphatidylinositol‐3‐kinase (PI3K)‐Akt pathway inhibit proinflammatory gene expression in macrophages. We determined whether the mammalian target of rapamycin (mTOR) and induction of the anti‐inflammatory cytokine interleukin‐10 (IL‐10) inhibited LPS‐induced proinflammatory and TF gene expression in peritoneal macrophages (PMs) in a PI3K‐Akt‐dependent manner. We used wild type (WT) peritoneal macrophages (PMs), and PMs from PTENflox/flox/LysMCre mice (PTEN−/− PMs), which have increased Akt activity. LPS induction of IL‐10 mRNA and protein was increased in PTEN−/− PMs compared to WT PMs. Pharmacologic inhibition of mTOR with rapamycin inhibited LPS induction of IL‐10 mRNA and protein, and enhanced the expression of TF and the proinflammatory cytokines interleukin‐6 (IL‐6) and tumor necrosis factor‐α (TNFα). Neutralization of endogenous IL‐10 also enhanced LPS induction of TNFα, IL‐6 and TF expression in WT and PTEN−/− PMs. The results indicate that mTOR‐dependent IL‐10 expression leads to inhibition of LPS induction of TF and the proinflammatory cytokines IL‐6 and TNFα in WT macrophages
Effects of Korean Chaga mushroom extract on stimulation of immune response in mouse splenocytes
Authors: Batjargal Enkhbaatar , Hong Heeok , An Jeung Cheol , Hwang Seong Gu
Published: June 2021

The Chaga mushroom (Inonotus obliquus) contains many compounds that were reported to promote immune system and antioxidative status of test animals. The present study was conducted to determine the antioxidant activity and effects of Chaga mushroom on proliferation, cell population and cytokine expression of splenocytes. Mouse splenocytes were treated with 1, 5, 10, 20 and 40 ppm of Chaga mushroom extract, and IL‐1 β, IL‐2 and TNFα cytokine expressions were determined after 24 and 48 h incubation. Cytokine expressions were measured by ELISA. Cell proliferation was measured by tetrazolium salt (WST) assay after 24 and 48 h incubation periods. The T cell and B cell populations of splenocytes were measured by Flow Cytometry Analysis (FACS). The free radical scavenging activity was measured using 2,2‐Diphenyl‐1‐picrylhydrazyl (DPPH) assay. The IL‐2 expression was stimulated by Chaga mushroom after 24 hours and 48 hours incubation periods. Splenocyte proliferation was stimulated in dose dependent manner by Chaga mushroom. The 24 and 48h data showed that T cells population was more stimulated than B cells. The DPPH assay showed that antioxidative activity of Chaga mushroom increased dose dependently with concentration. Results of this study suggested that Chaga mushroom promote immune response through improvement of antioxidation status and stimulation of IL‐2 and immune cell proliferation in animal.
Ontogeny of mRNA expression of pulmonary innate immune factors
Authors: Derscheid Rachel Jean , Gallup Jack , McCray Paul B , Ackermann Mark R
Published: June 2021

Prematurely born infants are at an increased risk of developing respiratory infections due in part to decreased function of their innate immune system. It was our goal to examine the ontogeny of innate immune gene expression by respiratory epithelia in our ovine model. Targets of interest are surfactant proteins A and D (SP‐A and SP‐D), sheep beta defensin 1 (SBD‐1), dual oxidases 1 and 2 (Duox1 and Duox2), lactoperoxidase (LPO), SPLUNC1, LPLUNC2, Toll‐like receptors 3, 7, and 8 (TLR 3, 7, and 8). Lung from sheep was collected at 115 and 130 days gestation, full term, 15 days post partum, and adulthood. Levels of mRNA for the target innate immune genes were measured by real‐time RT‐qPCR. LPO, SPLUNC1, LPLUNC2, TLR‐8, SBD‐1, and Duox1 expression increased progressively with age. Surfactant proteins A and D, previously reported to be decreased in preterm lambs, did not follow a steady trend, although SP‐A expression was lower at 130 days gestation than at full term. Duox2 levels were lower in gestational and perinatal lambs than in adults, but did not show a linear increase with age. These results are significant in that they may indicate that reduced expression in fetal lung may underlie susceptibility of the lung to infection with preterm birth. Conversely, upregulation of these genes may have therapeutic potential.
The anti‐platelet effects of apocynin are not mediated by inhibition of NADPH Oxidase
Authors: Dharmarajah Janahan , Arthur Jane , Sobey Christopher Graeme , Drummond Grant Raymond
Published: June 2021

The methoxy‐substituted catechol, apocynin is a powerful inhibitor of NADPH oxidase. Recently apocynin has been shown to prevent the aggregation of platelets in response to agonists such as collagen and thrombin. We examined whether NADPH oxidase plays a role in collagen‐induced aggregation, elucidating the mechanism(s) of action of apocynin on this enzyme complex. Platelets were isolated from 10 week‐old male C57BL6/J and Nox2‐deficient (Nox2−/−) mice to assess platelet aggregation (aggregometry) and NADPH oxidase expression (immunoblotting for Nox2). Collagen (1‐30 μg/ml) caused a concentration‐dependent increase in platelet aggregation (maximum response ~35.35 ± 3.18%; n=6). This response was inhibited by apocynin (P<0.05, n=6), albeit at higher concentrations (=100µM). Immunoblotting with an anti‐Nox2 antibody from platelets isolated from wild‐type mice revealed strong bands at 58kDa, 65kDa and 91kDa. Collagen was as effective at eliciting aggregation in platelets from Nox2−/− and wild‐type mice. Moreover, the superoxide scavengers, SOD (300U/ml) and PEG‐SOD (300U/ml) had no effect on platelet aggregation in response to collagen. These data suggest that while platelets express NADPH oxidase, it is not responsible for mediating collagen‐induced aggregation. Apocynin's anti‐platelet actions are likely to be due to an off target effect of the drug which remains to be determined. (NHMRC 384136)
Ultrafine particles increase the production of reactive oxygen species (ROS) and decrease contractile function in rat adult ventricular myocytes (ALVM)
Authors: Mahne Sarah Elizabeth , Shenouda Sylvia K. , Moll Alyson , Bilbo Corey , Lominicki Slawo , Dellinger Barry , Varner Kurt
Published: June 2021

Increases in the concentration of airborne fine particles (< 0.1 µm, UFP) are positively correlated with increased cardiovascular mortality. UFP produced by the combustion of fossil fuels and wastes contain stable free radicals capable of redox cycling and producing oxidative stress. After inhalation, UFP transit from the lung to the heart raising the possibility that UFP produces oxidative stress in the heart by a direct action. This study determined whether radical‐containing UFP produce oxidative stress and contractile deficits in ALVM. Rat ALVM were treated with SiCuO UFP containing dichlorobenzene (DCB230, 8.8 and 17.6 µg/cm2) for 24 hr. DCB230 increased ROS production (measured by dihydroethidium staining) in ALVM by 50% relative to control. The higher dose killed all the cells. An IonOPtix system was used to assess contractile function in treated and control ALVM. Compared to control, DCB230‐treated ALVM showed decreased sarcomere length shortening percent and decreased the maximal velocities of shortening and re‐lengthening during stimulation (1Hz). In conclusion, treatment with UFP directly increased ROS production and decreased contractile function in ALVM. Support RR018766
Mitochondrial membrane potential regulates production of reactive oxygen species and opening of mitochondrial permeability transition pore
Authors: Sedlic Filip , Wakatsuki Tetsuro , Pravdic Danijel , Bosnjak Zeljko
Published: June 2021

Anesthetic preconditioning (APC) protects the heart from ischemia/reperfusion injury. We hypothesized that APC by isoflurane protects the heart by attenuating generation of reactive oxygen species (ROS) that delays opening of mitochondrial permeability transition pore (mPTP) via mild decrease in mitochondrial membrane potential (Δψm). ROS and Δψm were measured in rat cardiomyocytes by real‐time confocal microscopy using TMRE and CM‐H2DCFDA fluorescent dyes, respectively. Opening of mPTP, which initiates cell death, was detected as rapid and complete loss of TMRE fluorescence from mitochondria. APC by isoflurane (0.5 mM) decreased TMRE fluorescence (88±5% of control) and increased flavoprotein fluorescence (109±3% of control), indicating decrease in Δψm and mitochondrial uncoupling, respectively. Low dose of protonophore DNP (100 nM) produced similar effect. Pyruvate (25 μM) increased respiration and reversed mitochondrial depolarization and uncoupling. When exposed to oxidative stress, myocytes with partly decreased Δψm produced less ROS (62±5% of control) and exhibited a delay in mPTP opening (162±7% of control). Increase in Δψm by pyruvate increased production of ROS and shortened the time of mPTP opening. In conclusion, APC protects cardiomyocytes in part via mild decrease in Δψm that attenuates ROS production under stress and leads to the delay in mPTP opening.

Supported by P01GM066730
Losartan reduces mortality in a model of heart failure
Authors: Neumann Joachim , Sophie Guenther , Jones Larry R , Hauptmann Steffen , Gergs Ulrich
Published: June 2021

Calsequestrin (CSQ) is the main Ca2+ store in the cardiac sarcoplasmic reticulum (SR). Overexpression of CSQ selectively in the heart of transgenic mice (TG) under the control of the alpha myosin heavy chain promoter leads to hypertrophy, fibrosis, heart failure, cardiac arrhythmias and ultimately premature death compared to littermate controls (WT). We hypothesized that losartan, an inhibitor of angiotensin II receptors, may be able to abrogate these detrimental effects. Hence, TG and WT were treated for about 2 or 5 months perorally with losartan (5 mg/Kg body weight) or solvent alone. Under control conditions (solvent alone), none of the WT died within the observation period whereas all TG died within 9 months. In the treatment group the mortality of TG was significantly reduced: mean life span was raised from 116 d to 192.5 d (n=18). Likewise, the degree of fibrosis was reduced. Hemodynamic parameters (pressure; dp/dt) were improved. However, the relative heart weight remained unchanged. Likewise, the expression of CSQ in the heart was unchanged by losartan treatment. In conclusion, the renin angiotensin system is apparently involved in heart failure due to CSQ overexpression, because its blockade improves survival (supported by the DFG).
Diacylglycerol ‐induced suppression of vascular KCNQ channels ‐A common mechanism of action of vasoconstrictor hormones?
Authors: Mani Bharath , Brueggemann Lioubov , Byron Kenneth
Published: June 2021

Vasoconstrictor hormones increase cytosolic Ca2+ concentration in vascular smooth muscle cells (SMC) by binding to receptors that activate phospholipase C, resulting in production of diacylglycerol (DAG). DAG is a known activator of protein kinase (PKC) and a direct activator of some non‐selective cation channels. In A7r5 rat aortic SMC, we had previously shown that Arg8‐vasopressin (AVP), induces repetitive Ca2+ transients (Ca2+ spiking) by PKC‐dependent suppression of KCNQ potassium channels. We examined the role of DAG using fura‐2 Ca2+ measurements and electrophysiological recordings in A7r5 cells. A DAG analogue, OAG (100µM), inhibited KCNQ currents by 56% at ‐20mV. OAG also activated a non‐selective cation current (ICAT) and stimulated Ca2+ spiking. Inhibition of PKC using 2µM Ro‐31‐8220 completely abolished OAG‐induced Ca2+ spiking. TRPC6 knockdown (by expression of TRPC6 shRNA) abolished ICAT but only modestly delayed the onset of AVP‐ or OAG‐induced Ca2+ spiking. These results suggest that while activation of TRPC6 channels contributes to AVP‐induced Ca2+ spiking, DAG‐induced suppression of KCNQ channels is the primary mediator of the response. Another vasoconstrictor hormone serotonin (10nM) also inhibited 94% of KCNQ current and stimulated Ca2+ spiking. Hence DAG‐mediated suppression of KCNQ channels might be a common mechanism by which vasoconstrictor hormones initiate Ca2+ responses.
Stroke related alterations in protease activated receptor‐2 and bradykinin mediated vasodilation in the middle cerebral arteries of stroke prone hypertensive rats
Authors: Smeda John Stanislaw , McGuire John , Daneshtalab Noriko
Published: June 2021

Vasodilation to bradykinin (1.2 µM) and protease‐activated receptor‐2 activation by 2‐furoyl‐leucine‐isoleucine‐glycine‐arginine‐leucine‐ornithine‐amide (2‐FLY, 1 µM) was measured in isolated pressurized (100 mmHg) middle cerebral arteries (MCAs) from Kyoto Wistar stroke prone spontaneously hypertensive rats (SHRsp) before and after hemorrhagic stroke (HS). Vasodilation to both peptides was endothelial dependent and produced by a non NO, cyclo‐oxygenase dependent mechanism that activated endothelial IKCa+SKCa channels and promoted vasodilation through smooth muscle BKCa, Kir channels and Na+/K+ ATPase activation. After HS, MCAs lacked the ability to vasodilate to bradykinin. Vasodilation to 2‐FLY was not altered. Post stroke oral Losartan (35 mg/kg/day) or Captopril (50 mg/kg/day) treatment did not reduce blood pressure. Losartan treatment restored MCA bradykinin vasodilation after 7 days for the duration of the study (>100 days treatment). Captopril only temporarily restored bradykinin relaxation between 7 to 25 days of treatment after which this function deteriorated. HS produced selective alterations in MCA endothelial function that could not be explained solely by the level of hypertension or as a secondary consequence of HS. These dysfunctions were reversed more effectively in SHRsp after HS by blocking AT‐1 receptors as opposed to lowering angiotensin II levels.
ERK activation by the alpha1‐adrenergic receptor agonist phenylephrine in human aortic smooth muscle (AoSMC) cells
Authors: Norman Derek D. , Cole Judith A.
Published: June 2021

The adaptive process of vascular remodeling causes structural alteration in blood vessels via proliferation, differentiation and migration of vascular smooth muscle cells (VSMCs) within the vessel wall. These changes play roles in both normal vessel activity and the appearance of pathophysiological conditions such as hypertension and atherosclerosis. ERK plays a role in all of these activities, and α‐adrenergic receptors (αARs) activate ERK by classic second messengers and by EGF receptor (EGFR) transactivation. Using a phosphoantibody cell‐based ELISA, we measured ERK activation by α1 and α2AR agonists. While AoSMCs express both α1 and α2ARs, ERK was activated only by the α1 agonist phenylephrine (PE). Surprisingly, the α1 agonist cirazoline had no effect on ERK activity. ERK activation by PE was blocked by the α1 antagonist prazosin and the α1A antagonist SNAP 5089 but was not affected by the α1D antagonist BMY 7378. While EGFR downregulation reduced ERK activation by PE, the EGFR inhibitor PD168393 did not. Further study will be necessary to elucidate the exact role of the EGFR in PE‐mediated ERK activation; however, our data show that activation of ERK by PE occurs in a potentially EGFR‐dependent process via activation of α1A but not α1DARs. Ultimately, we wish to determine αAR subtype‐specific, ERK‐mediated expression of immediate early genes which play a role in the processes involved in vascular remodeling.
Biophysical characterization of Gβγ “hot spot” binding small molecules: explaining Gβγ “hot spot” binding effector selectivity
Authors: Seneviratne Pramodh , Wedekind Joseph , Burroughs Michael , Smrcka Alan
Published: June 2021

Guanine nucleotide exchange proteins (G proteins) are heterotrimeric proteins composed of three subunitsα, β, γthat initiate intracellular signaling cascades. Previously our laboratory identified small molecules that bind to a protein‐protein interaction "hot spot" to achieve selective modulation of downstream effectors. We hypothesized that different compounds bind directly to different sub‐surfaces of the Gβγ 'hot spot'. To assess binding of compounds directly to Gβγ we developed a Surface Plasmon Resonance (SPR) method utilizing immobilized Gβγ subunits. The results indicate that multiple compounds bind directly to Gβγ with affinities in the range of 400 nM to 3μM. For example: the Kd for binding of Gallein to Gβ1γ2 was 420 nM with kon=1 x 103 M−1s−1, koff= 4 x 10−4 s−1. We separately purified Gβγ subunits with single amino acids within the Gβγ hotspot mutated to alanine and have obtained unique binding maps for 3 small molecules using SPR. To corroborate our SPR binding maps we also carried out binding experiments using 3H labeled Gallein. We are also in the process of obtaining co‐crystals of compounds bound to Gβγ, and solving their 3D structures using x‐ray crystallography. Understanding how selective small molecules interact with Gβγ will be important for determining the mechanism of action of these compounds and developing more potent and selective agents.

This work was funded by (NIH GMRO1‐081772)
An age‐related deficit in working memory emerges in the Ts65Dn mouse, a murine model of Down syndrome
Authors: Whitney Katharine N , Wenger Galen R
Published: June 2021

Previous research showed an initial deficit in working memory in young adult, male Ts65Dn mice compared to young adult, male littermate controls (LC). This deficit disappeared with additional training (Dowdy‐Sanders and Wenger, Behav. Brain Res. 168: 349‐352, 2006), suggesting a delayed learning curve rather than a true deficit in working memory. The purpose of this study was to determine the effect of age on working memory in male Ts65Dn and LC mice responding under a matching‐to‐position schedule of reinforcement. The mice were trained beginning at 3‐ months of age, and were tested until 24‐ months of age. Similar to the prior study, there was an initial deficit in the working memory of the Ts65Dn mice compared to LC that disappeared with practice. Once the initial deficit was reduced by additional training, the length of the mean delay for the session achieved by the two lines of mice was indistinguishable, and remained stable until after 20‐ months of age. Between 20‐ and 24‐months of age a difference in the mean delay for the session emerged between the two lines. These results suggest a difference in the cognitive decline of aged Ts65Dn mice compared to aged LC mice. Supported by NIH grant HD047656 (G.R.W.)
The tobacco smoke constituent, norharmane enhances nicotine self‐administration in an age‐dependent manner
Authors: Arnold Monica Martin , Prenger Jacque A , Belluzzi James D
Published: June 2021

Smoking is the most preventable cause of death in the United States. Twenty‐one percent of U.S. adults consider themselves smokers with most beginning when they were teenagers. Although numerous smokers report the desire to stop, quit rates are low and current cessation aides are minimally effective. Animal models created to study smoking generally only use nicotine, the main psychoactive component of smoke; however experimental results do not reflect the addictiveness of cigarettes. This discrepancy may be because adult animals are used whereas initiation begins in adolescence, a critical time of brain development. Moreover, only nicotine is used despite that there are over 4000 constituents found in cigarette smoke. Therefore, the present study investigated how the age of acquisition and the tobacco constituent, norharmane, may influence nicotine self‐administration in adult and adolescent male rats. The co‐administration of nicotine (7.5 µg/kg/inj) and norharmane (0.1‐2.5 µg/kg/inj) at doses equivalent to what is found in the plasma of smokers after smoking one cigarette enhanced nicotine self‐administration in naïve adults compared to nicotine alone. Adolescents were insensitive to any of the drug combinations and responded minimally overall. Consequently, we have found that there is an age‐dependent sensitivity in responding for nicotine alone and in combination with norharmane.

Sponsored by NIH DA 21267 & 19138.
Characterization of PG‐619, a dopamine D3 receptor partial agonist, on cocaine self‐administration and drug‐elicited yawning in rhesus monkeys
Authors: Blaylock Brandi L , Gould Robert W , Newman Amy H , Nader Michael A
Published: June 2021

The dopamine D3 receptor is a promising target for the development of novel treatment agents for addiction due to its restricted expression in limbic brain regions and involvement in reinforcement pathways (Newman et al., 2005). The present study examined the effects of PG‐619, a D3 receptor partial agonist, with 100‐fold selectivity for the D3 over D2 receptors (Grundt et al., 2007), on cocaine self‐administration (SA) and yawning in male rhesus monkeys with an extensive cocaine history (n=3). Monkeys self‐administered cocaine (0.003‐0.3 mg/kg) under a fixed‐ratio 30 schedule during daily 2‐hr sessions. Prior work found CJB 090, a D3 partial agonist with 60‐fold D3 selectivity, decreased cocaine‐maintained responding (Martelle et al., 2007). Acute doses of PG‐619 (0.1 ‐ 1.0 mg/kg, i.v., 10 min presession) did not significantly affect cocaine SA; higher doses of PG‐619 are currently being evaluated. In vivo potency of PG‐619 to elicit yawning was also examined. As with CJB 090, no doses of PG‐619 (0.1 ‐ 1.0 mg/kg) elicited yawning. The preliminary findings support prior work that D3 partial agonists do not have D3 agonist effects in vivo. Furthermore, the findings suggest greater selectivity of D3 over D2 receptors may result in substantial decreases in potency to affect cocaine SA, although bioavailability and pharmacokinetic factors may contribute to the behavioral profiles of these drugs. DA 12460 and NIDA‐IRP.