bluuebunny/crossref_metadata_2025 huggingface dataset:
Gut fermentation and health effects of Louisiana sweet potato varieties
Authors: McCutcheon Kathleen Lee , LaBonte Don R , Picha David H , Williams Cathleen C , Keenan Michael J , Martin Roy J
Published: June 2021

Sweet potatoes (SP) are high in phytonutrients and a healthy alternative to white potatoes (WP). Both are naturally low in calories and fat, high in vitamins, minerals and fiber with similar carbohydrate starch, sugar and fiber profiles, but preparation methods affect starch digestion in WP. Thus, 20% of carbohydrates in SP are resistant starches (RS) that slows digestion and the release of sugar. RS is fermented by microflora to produce short chain fatty acids (SCFA), affect peptide secretion, and improve absorption. New SP varieties developed for disease and pest resistance by the LSU AgCenter's Sweet Potato Research Station were tested in vivo to determine health benefits when included in a daily diet. For 12 weeks, rats were fed daily a purified diet with freshly baked potatoes as the carbohydrate source (2 orange flesh (O), 2 white flesh (W) SP varieties and a control WP). All SP groups had significantly less body fat with variety differences between WSP (lowest body fat) and OSP (lowest pH). Daily intake of SP can increase health benefits from gut fermentation as indicated by significantly lower cecal pH, larger cecum weights and greater production of total SCFA in all SP fed groups.
Partial substitution of fructan fibers with acacia gum altered fermentation profile in an in vitro batch system fermentation
Authors: Noack Jacqueline , Klosterbuer Abby , Timm Derek , Thomas William , Slavin Joanne
Published: June 2021

Fructan fibers are added to enteral formulas for their prebiotic benefits and production of short chain fatty acids (SCFA), yet their rapid fermentation may lead to excess gas and gastrointestinal intolerance. We hypothesized that partial substitution of fructan fibers with slowly fermented acacia gum (AG) would decrease gas production while preserving SCFAs. Two blends of equal amounts of total fiber were compared using an in vitro batch fermentation model. Blend 1 contained FOS and inulin. Blend 1+ contained FOS, inulin, and AG. Triplicate samples were removed at 4, 8, 12, and 24 h for analysis of gas, pH, and SCFAs. In general, Blend 1 produced more gas earlier than Blend 1+. Blend 1 produced 5 times the gas at 8 h (P < 0.05), while Blend 1+ produced 1.2 times the gas at 24 h (P < 0.05). Blend 1+ took longer to reach its maximum rate of gas production (11.13 ± 0.93 h), compared to Blend 1 (4.7 ± 0.06 h). Blend 1 had more acetate, butyrate, and total SCFAs at 4 h, while Blend 1+ had more propionate at 8 h, butyrate at 12 h, and acetate at 24 h (P < 0.05). In conclusion, partial substitution of fructan fibers with AG produced less gas early on while maintaining production of SCFAs. These results may help predict health benefits and gastrointestinal tolerance in vivo.

Research was supported by Nestlé Health Science.
Postprandial effect of a novel rice product on blood glucose in healthy men
Authors: Poquette Nicole , Arijaje Emily , Wang Ya-Jane , Lee Sun-Ok
Published: June 2021

The objective of this human study was to investigate the effect of a novel parboiled brown rice product on blood glucose levels. We hypothesized that the prepared‐parboiled brown rice product caused lower blood glucose responses than the control glucose solution. Using a randomized crossover design, 14 healthy male participants were divided among 3 cohorts (4–5 subjects per cohort) and consumed both treatments with a one‐week washout period. After consuming the rice product, subjects showed significantly lower blood glucose response at 15, 30, 45, 60, 75, and 90 minutes in comparison with the control glucose solution (P<0.05). Subsequently, the incremental AUCs (Area Under the Curve) of blood glucose were also significantly lower with the novel rice product (3795±602 mg·(−15~180 min)/dL) than the control glucose solution (5880±658 mg·(−15~180 min)/dL) (P<0.05). In addition, the incremental AUCs of plasma insulin response reduced significantly to 37–40% when subjects ingested the rice product comparing with the glucose solution (P<0.05). Results suggest the novel brown rice product would assist in managing blood glucose levels for individuals suffering from diabetes. Funding provided by Arkansas Biosciences Institute is greatly appreciated.
How analysis of data from alpha‐amylase catalysed starch digestibility performed in vitro contributes to an understanding of rates and extent of digestion starchy foods in vivo
Authors: Butterworth Peter J. , Warren Frederick J. , Edwards Cathrina H. , Grassby Terri , Patel Hamung , Ellis Peter R.
Published: June 2021

Ingestion of different foods containing identical amounts of starch can result in very different postprandial rises in blood glucose and insulin concentrations. Limitation of the early rises in blood glucose and insulin levels seems to be beneficial to human health in the long term. Many studies of starch digestion in vitro are made to understand the molecular basis for differences in digestion rates in vivo to enable prediction of likely rates of digestion of particular starchy foods. Michaelis‐Menten kinetics of starch digestibility provides estimates of available (digestible) substrate as starch samples are hydrothermally treated. Combined with studies of starch structure using calorimetry and FTIR spectroscopy, key features influencing rates of amylolysis were identified. Measurement of product formation during prolonged incubations of starch with α‐amylase produces digestibility curves. Use of logarithm of slope (LOS) plots to analyse the curves by 1st order kinetics gave values for digestibility rate constants and the total digestible starch, C∞. An important conclusion is that contrary to many reports in the literature, cooked starches do not contain distinct fractions of rapidly and slowly digested material. These kinetic approaches have also been used in studies of plan‐tencapsulated starch to understand how cell walls influence access of amylase to starch. This work was funded by the BBSRC, UK (DRINC BB/H004866/1).
Variability of Vitamin A and Carotenoid Status Among Women of Childbearing Age
Authors: Goodman Michael J , Murtaugh Maureen A
Published: June 2021

Objective

Animal models suggest vitamin A exposure at subteratogenic levels may contribute to behavioral problems in offspring. We aimed to determine whether serum levels of retinol retinyl palmitate and carotenoids were higher among potentially fertile supplement users.

Methods

100 women completed a dietary history questionnaire. Blood samples were analyzed for retinyl palmitate, α‐carotene, β‐carotene, lutein, and zeaxanthin. Pearson correlations between reported intake and serum levels were calculated, and serum levels were compared among supplement users and non‐users.

Results

Mean levels and IQR of retinols and carotenes were: retinyl palmitate 0.019 (0.10–0.20), vitamin A 0.54 (0.45–0.59), α‐carotene 77.15 (36.5–95.5), β‐carotene 228.34 (94.00–298.50), zeaxanthin 20.29 (12.00–26.50). 31 women took multi‐vitamins containing vitamin A (mean 4332 IU). Serum levels of retinol and carotenoids were highly variable. We observed elevated levels of retinol (n=1) α‐carotene (n=21) level, β‐carotene (n=14), and lutein (n=5). Serum β‐carotene was correlated with β‐carotene (mg/d) from supplements (r= 0.21, p = 0.03).

Conclusion

Vitamin A and carotenoid status of potentially fertile women was variable. Serum β‐carotene levels were associated with supplement use whereas retinol levels were not distinguishable between users and non‐users.
Coconut oil facilitates tomato carotenoid tissue accumulation in the Mongolian gerbil (Meriones unguiculatus)
Authors: Conlon Lauren E , King Ryan D , Moran Nancy Engelmann , Erdman John W
Published: June 2021

Tomato carotenoids have demonstrated anti‐cancer bioactivity, and co‐consumption of carotenoids with a polyunsaturated fat source has been shown to enhance bioavailability. To investigate the effect of fatty acid chain length and saturation on tomato carotenoid tissue accumulation, Mongolian gerbils (Meriones unguiculatus) were fed a 20% fat diet. Gerbils (n = 40) were fed control diets with safflower or coconut oil, or experimental diets containing 10% tomato powder and coconut oil or 10% tomato powder and safflower oil for 28 days. Coconut oil increased carotenoids among many compartments including total carotenoids in the serum (p = 0.0001), adrenal glandular phytoene (p = 0.047), hepatic phytofluene (p = 0.0001), testicular all‐trans lycopene (p = 0.01), and cis‐lycopene (p = 0.006) in the prostate‐seminal vesicle complex compared to safflower oil. Safflower oil‐fed gerbils had greater splenic lycopene (p = 0.006) compared to coconut oil fed gerbils. Coconut oil also increased serum cholesterol (p = 0.0001), and decreased hepatic cholesterol (p = 0.0003) compared to safflower oil suggesting a shift in the carotenoid‐rich LDL cholesterol flux that may have facilitated extrahepatic carotenoid tissue deposition. In summary, dietary fatty acid profiles may impact both lipid metabolisms and tissue carotenoid deposition.

Grant Funding Source : Funded in part by the Khatchaturian family
Discovery of a novel cytosolic ferroxidase in rodent enterocytes
Authors: Lu Yan , Ranganathan Perungavur N. , Collins James F.
Published: June 2021

During iron absorption in the doudenum, iron that exits enterocytes must be oxidized by a membrane bound multi‐copper ferroxidase, hephaestin. We designed experiments to assess Heph activity during iron deficiency and made the unexpected observation that ferroxidase (FOX) activity (by a transferrin‐coupled assay) was present in the cytosol of isolated rodent enterocytes. Several different experimental approaches allowed us to conclude that cytosolic FOX (cytoFOX) activity was not due to contamination with membrane Heph. cytoFOX activity was inhibited by sodium azide (a known FOX inhibitor), pPD (a competing substrate), D‐penicillamine (a copper chelator), and by SDS and heat (showing that it is mediated by a protein). Moreover, activity was increased (~30%) in iron deficient rats but was unchanged in copper deficient rats (in contrast to the reported dramatic reduction of Heph during copper deficiency). Finally, studies done in Heph KO, sla (Heph mutant) and ceruloplasmin (Cp) KO mice proved that cytoFOX activity was not contributed by Heph (or Cp) as activity in enterocyte cytosol was not different between KO/mutant mice and wt littermates. In conclusion, rodent enterocytes contain a cytosolic FOX that may function in transepithelial iron transport, perhaps acting in acidified intracellular vesicles containing the iron exporter ferroportin 1 (Fpn1) and complementing membrane Heph.

Grant Funding Source: NIH
Low prevalence of iron deficiency and iron deficiency anemia in children attending daycare in a large Canadian city
Authors: Jean-Philippe Sonia , Pham Thu , Hazell Tom , Vanstone Catherine , Agellon Sherry , El Hayek Jessy , Weiler Hope
Published: June 2021

Children are vulnerable to iron deficiency (ID) and iron deficiency anemia (IDA) due to rapid tissue expansion. Recent Canadian surveys reported that 1–3 y and 4–8 y in Québec had an iron intake of 10.1 mg and 13.7 mg/day respectively; however, iron status was not assessed. This study examined the prevalence of ID and IDA and iron intake in children (n=523) from randomly selected daycares in Montréal. Ferritin was measured using an autoanalyzer (Liason®, DiaSorin) and hemoglobin using a radiometer (ABL80 FLEX Radiometer Medical A/S) in capillary samples. Iron intake over 24 h for 469 children with 72 having a repeat assessment was analyzed using Nutritionist Pro™ (Axxya Systems LLC.) and the Canadian Nutrient File. The data was adjusted for day‐to‐day variation, and all children met the EAR (3 mg/day for 1–3 y, 4 mg/day for 4–5 y olds). Mean daily intake for these age groups was 10.22±5.70 mg (n=272) and 11.06±5.32 mg (n=197), respectively, was similar to national data. ID (ferritin <12 μg/L) was present in 13.4% of children and IDA (hemoglobin <110 g/L) in 2.9% of this population. Corresponding means were 22±13 μg/L and 126±19 g/L. This data complements our recent cross‐sectional study reporting ID in 18.0% and IDA in 5.4% in 3–5 y Inuit children; confirming that urban dwelling preschoolers in a large Canadian city have a lower prevalence of ID and IDA. Supported by the Beef Information Centre.
Mathematical Modeling of Iron Uptake by Tim‐2 Receptor in Mouse Kidney Cells
Authors: Han Jian , Xie Zhijian
Published: June 2021

Iron is an essential element in all living cells. Classically, cells acquire iron through transferrin‐transferrin receptor pathway. In addition to this pathway, iron can be taken up through H‐ferritin (HFt) binding to Tim‐2 receptor in mice. In this research, the iron uptake process through Tim‐2 was computed by mathematical models to illustrate the dynamics of iron uptake, storage, and export in mice kidney cells. Simulation results showing iron metabolic process by Tim‐2 pathway were compared with biological experimental data done in vitro. The model predicts the fates of HFt and iron in Tim‐2 pathway and produces equations to calculate rate of iron flow within different cellular compartments. The model predicts desirable characteristics for intracellular iron homeostasis and reveals that the rate of iron storage in endogenous intracellular ferritin is around 2.8% per minute, and export rate through non‐Tim2 protein is about 1.0% per minute. The model incorporates three iron metabolic experiments in different time scale under one consistent framework. The model captures the complexity of iron metabolism and connects several metabolic processes into one picture for easy understanding. The model can be applied to general iron metabolism in other cell types.
High dietary iron and radiation exposure increase biomarkers of oxidative stress in blood and liver of rats
Authors: Morgan Jennifer L. L. , Theriot Corey A. , Wu Honglu , Smith Scott M. , Zwart Sara R.
Published: June 2021

Radiation exposure and increased iron (Fe) status independently cause oxidative damage that can result in protein, lipid, and DNA oxidation. During space flight astronauts are exposed to both increased radiation and increased Fe stores. Increased body Fe results from a decrease in red blood cell mass and the typically high Fe content of the food system. In this study we investigated the combined effects of radiation exposure (0.375 Gy of Cs‐137 every other day for 16 days; a total of 3 Gy) and high dietary Fe (650 mg Fe/kg diet vs. 45 mg Fe/kg for controls) in Sprague‐Dawley rats (n=8/group). Liver and serum Fe were significantly increased in the high dietary Fe groups. Likewise, radiation exposure increased serum ferritin and Fe concentrations. These data indicate that total body Fe stores increase with both radiation exposure and excess dietary Fe. Hematocrit decreased in the group exposed to radiation, providing a possible mechanism for the shift in Fe indices after radiation exposure. Markers of oxidative stress were also affected by both radiation and high dietary Fe, evidenced by increased liver glutathione peroxidase (GPX) and serum catalase as well as decreased serum GPX. Our results reveal preliminary indications of the synergistic effects of protracted (fractionated) radiation exposure and increased dietary Fe, warranting further study.

Grant Funding Source: This research was funded by the NASA Human Research Program. Dr. Morgan's efforts and travel are supported through an appointment to the NASA Postdoctoral Program at the Johnson Space Center, administered by Oak Ridge Associated Universities through a contract with NASA
Ratio of primary to secondary oxidation products and time of exposure correlate with lipid accumulation in murine adipocytes
Authors: Skinner Joanna Ponder , Dingels Nicole , Santanam Nalini , Penumetcha Meera
Published: June 2021

The objective of this study was to investigate if the level of oxidation of soybean oil determines the accumulation of lipids in 3T3‐L1 adipocytes.

Methods

Soybean oil (SO) was unheated or heated for 3, 6 or 9 hours at 190°C to generate oils with increasing levels of oxidation as determined by the levels of conjugated dienes (CD) and conjugated trienes (CT). 3T3‐L1 cells were grown to confluence in 24 well plates and then exposed to DMEM supplemented with or without 0.1%, 0.01% and 0.001% of heated or unheated SO for 24 hrs either prior to or, after differentiation. After treatment, cells were stained with oil red O to determine the accumulation of lipids.

Results

Progressive heating of soybean oil created oils with increasing amounts of oxidized lipids (ratio of CD/CT was 1.92, 2.33, 3.1 and 3.56 for the unheated, 3hr, 6hr and 9hr heated oils, respectively). The intensity of oil red O staining was similar in 3T3‐L1 cells that were treated with vehicle or unheated oil for 24hr in undifferentiated cells but was lesser in the cells treated with 3hr heated SO. More interestingly, there was no oil red staining when cells were treated with the 6hr and 9hr heated oils, suggesting a loss of lipid accumulation. In experiments where SO was added to differentiated 3T3‐L1 cells for 24 hrs, there was no obvious effect of the oils on oil red O staining.

Conclusion

These results demonstrate that exposure of pre‐adipocytes to oils with higher ratios of oxidized lipids inhibits the adipocyte lipid accumulation. Since this effect is not seen in cells that are differentiated, it suggests that heated oils could be suppressing regulators of adipocyte differentiation.

Grant Funding Source : None
Development of a flavonoid database for assessing population exposures and its application
Authors: Ahuja Jaspreet KC , Bhagwat Seema , Haytowitz David , Holden Joanne , Bailey Reagan , Dwyer Johanna , Milner John , Moshfegh Alanna
Published: June 2021

Flavanoids are a family of polyphenolic compounds found in foods of plant origin and are of interest because of their potential role in health and disease prevention. Many knowledge gaps exist in the study of flavonoids, including the lack of a comprehensive food composition database so assessing population exposure is difficult. To address this need, flavonoid data from the USDA Database for the Flavonoid Content of Selected Foods, 3.0 (2011) was applied to over 7,000 foods in the Food and Nutrient Database for Dietary Studies, 4.1, the database used for analyzing dietary intakes in What We Eat in America (WWEIA), NHANES. The USDA Database for the Flavonoid Content of Selected Foods, 3.0 has been expanded to include 26 aglycone compounds in 5 subclasses – flavonols, flavones, flavanones, flavan‐3‐ols, & anthocyanidins, & 3 isoflavones compounds in 3,000 foods. Further, the database was applied to food intake data from 8,529 males and females, ages 2 year & older from WWEIA, NHANES 2007–2008 to determine major foods sources of individual classes of flavonoids in the U.S. population. For example, onions and apples are among the important food sources of the flavonol ‐ quercetin. The database provides researchers for the first time with a tool to assess intake patterns of individual flavonoid compounds, identify foods that are major contributors, and investigate diet‐disease relationships in the U.S. population.
A‐type procyanidin dimers, trimers, and tetramers from cranberries transported across monolayers of human Intestinal epithelial Caco‐2 cells
Authors: OU Keqin , Percival Susan S. , Zou Tao , Khoo Christina , Gu Liwei
Published: June 2021

A‐type procyanidin oligomers in cranberries are known to inhibit the adhesion of uropathogenic bacteria. B‐type procyanidins dimers and trimers are absorbed by humans. The absorption of A‐type procyanidins from cranberries in humans has not been demonstrated. This study examined the transport of A‐type cranberry procyanidin dimers, trimers, and tetramers on differentiated human intestinal epithelial Caco‐2 cell monolayers. Procyanidins were extracted from cranberries and purified using chromatographic methods. Fraction I contained predominantly A‐type procyanidin dimer A2 [epicatechin‐(2‐O‐7, 4‐8)‐epicatechin]. Fraction II contained primarily A‐type trimers and tetramers, with B‐type trimers, A‐type pentamers, and A‐type hexamers being minor components. Fraction I or II in solution were added onto the apical side of the Caco‐2 cell membranes. The media at the basolateral side of the membranes were analyzed using HPLC‐MSn after two hours. Data indicated that procyanidin dimer A2 in fraction I and A‐type trimers and tetramers in fraction II traversed across Caco‐2 cell monolayers with transport ratio of 0.59%, 0.37%, and 0.23%, respectively. This study demonstrated A‐type dimers, trimers, and tetramers were transported across Caco‐2 cells at low rates, suggesting they could be absorbed by humans after cranberry consumption.

This research was supported in part by Ocean Spray Cranberries, Inc.
Effect of vitamin D on aging in Caenorhabditis elegans
Authors: Messing Jennifer A. , Schisa Jennifer A , Heuberger Roschelle A
Published: June 2021

Nutrigenomics, also referred to as nutritional genomics, aims to discover how nutrients interact with the genome and potentially alter genetic expression. One nutrient of particular interest is vitamin D3. In addition to its role in bone health, recent literature suggests a role for vitamin D3 in the prevention of various cancers, such as colon, prostate, and breast cancer, as well as the prevention of multiple chronic diseases, including cardiovascular disease, multiple sclerosis, type 1 diabetes mellitus, and rheumatoid arthritis. Based on its extensive actions within the body, vitamin D3 supplementation is sometimes warranted. We are using Caenorhabditis elegans as a model organism to study the effect of vitamin D3 on aging. In addition to the fact that this nematode has a relatively short lifespan, the DAF‐12 nuclear hormone receptor in C. elegans is homologous to the Vitamin D Receptor (VDR) in humans. Our hypothesis was that vitamin D3 supplementation would increase the lifespan of Caenorhabditis elegans as a result of vitamin D3's interaction with the DAF‐12 receptor. A dose‐response curve was completed to identify an effective concentration of vitamin D3, and our preliminary results indicate that the vitamin D3 significantly increases lifespan at a concentration of 1000 μg/ml. We are also in the process of comparing the longevity of wild‐type worms and daf‐12 mutants after exposure to vitamin D3. Results from these on‐going experiments will be presented. Our studies appear to be the first to utilize C. elegans as a model system to study vitamin D3. We anticipate that our work will demonstrate the utility of this approach, and that others may find this model system beneficial in nutrigenomic studies. This research was supported by an NIH grant to J. Schisa.
Effect of maternal soy protein isolate consumption on hepatocarcinogenesis in adult rat offspring
Authors: Kwon Young Hye , Choi Jihye
Published: June 2021

Maternal nutrition has been reported to program the offspring's susceptibility to later development of chronic diseases. Because we have observed increased liver proliferation in 3 week old offspring of dams fed a soy protein isolate (SPI) diet compared with offspring of dams fed a casein (CAS) diet, we investigated effects of maternal diets differing in protein source on diethylnitrosamine (DEN)‐induced hepatocellular carcinoma (HCC) in adult offspring. Female SD rats were fed a casein diet (CAS) or a low‐isoflavone soy protein diet (SPI) for two weeks before mating and throughout pregnancy and lactation. Offspring were weaned to a chow‐diet throughout the study. To induce HCC, offspring were received i.p. injection of DEN once a week for 14 weeks. We did not observe any nodules in offspring injected with saline. Multiplicity of nodules increased in SPI/DEN group compared with CAS/DEN group. Consistently, we observed higher incidence of adenomas and HCCs in SPI/DEN group. There was no difference in hepatic damage as measured by serum ALT activity, although tumor necrosis factor α levels were higher in SPI/DEN group. Based on histological examination, difference in liver necrosis, inflammation, steatosis, and fibrosis was not observed between groups. Therefore, maternal nutrition may program DEN‐induced hepatocarcinogenesis in adult offspring. This work was supported by National Research Foundation of Korea.
Child nutritional status and cognitive performance in Hawassa Town, Southern Ethiopia
Authors: Girma Meron , Loha Eskindir , Bogale Alemtsehay , Stoecker Barbara J
Published: June 2021

Relationships between anthropometry, diet diversity and cognitive performance were assessed in a cross‐sectional study of school children (n=116) aged 7–9 years. Participants were selected by two stage random sampling. Socioeconomic characteristics, dietary patterns, and anthropometrics were measured. The Raven's Colored Progressive Matrices (CPM board version) and selected tests from the Kaufman Assessment Battery for Children (KABC‐II) were used to assess cognitive performance. Mean (SD) weightfor‐ age (WAZ), height‐for‐age (HAZ) and BMI‐for‐age Z scores were −0.85 (0.94), −0.68 (1.13) and −0.62 (0.80) respectively. Mean (SD) diet diversity score was 3.8 (1.1). Stunted (HAZ < −2 Z) children had significantly (p<0.05) lower cognitive test scores compared to children not stunted for Raven's CPM, Number Recall and Pattern Reasoning. Underweight children (WAZ < −2 Z) had significantly lower CPM scores compared to children not underweight. Children who consumed meat, poultry, fish or eggs in the 24 hrs preceding the survey had significantly higher scores for Number Recall, Word Order and Triangles. Height‐for‐age and maternal education were significant predictors of Pattern Reasoning test scores (Adj. R2=0.182). Child diet diversity and anthropometric status and maternal education support cognitive function. (Supported by Hawassa University School of Graduate Studies and NIHR01HD053053 [NICHD & Fogarty International Center]).
Food security, nutrition and ART adherence in Latin America
Authors: Martinez Homero , Lynemmayr Sebastian , Ramírez Blanca Yohisy , Farías Hugo , Adams Jayne , Derose Kathryn Pitkin , Smith Alexandria
Published: June 2021

To develop a culturally appropriate, multi‐component nutrition intervention for people living with HIV in Honduras, and test its effect on ART adherence. 401 adults receiving ART who showed household food insecrity and/or undernutrition were recruited from four HIV clinics received nutritional assessment and dietary counseling, and were followed at 0 and 6 months to assess adherence (self‐report, pharmacy and clinic records). Randomly, two clinics (intervention) were selected to also receive a supplemental household food basket. At baseline, overall, more than two‐thirds missed at least one clinic appointment; over half did not pick up their pharmacy refills on time or reported missed taking their medicine, and; only about half had normal body mass index (BMI). After 6 months 11% of intervention group (both genders) changed BMI to normal, vs. 2% in control group, more evident in men (20% in intervention group improved BMI vs. 0% in the control). 80% of the intervention group showed improvements in coming to the clinic without fail, vs. 50% in control group, and 50% in the intervention group were more timely in picking up their refills on time vs. 25% in the controls. Improvements in nutritional status were significantly correlated with adherence measures (timely drug refill, not stop taking medicine). Our results underscore the importance of nutritional support for people in HIV care for Honduras and other countries.
Recombinant Mojastin Disintegrins Inhibit Cell Proliferation and Migration of SK‐Mel‐28 Cells and Migration of HT‐144 Cells
Authors: Bella Terri D. , Gutierrez Daniel A. , Bueno Raymund , Soto Julio G.
Published: June 2021

Disintegrins are low molecular weight peptides derived from viper venom. Three recombinant (r) mojastin peptides (Moj‐WN, Moj‐ MP, and Moj‐DM) were tested for their ability to inhibit migration and proliferation of SK‐Mel‐28 cells. Additionally, the r‐Moj‐DM peptide was tested for its ability to inhibit cell migration of HT‐144 cells. We hypothesize that r‐Moj‐DM and r‐Moj‐MP will inhibit cell migration and proliferation of SK‐Mel‐28; and r‐Moj‐DM will inhibit cell migration of HT‐144 cells. At 3μM, all three r‐Moj peptides inhibited cell proliferation of SK‐Mel‐28. The r‐Moj‐MP and r‐Moj‐DM peptides were the strongest inhibitors, preventing cell proliferation at 69% and 66%, respectively, compared to 28% inhibition by r‐Moj‐WN. At 3μM, r‐Moj‐WN failed to inhibit SKMel‐ 28 cell migration at significant levels as compared to the untreated controls (p=0.108). At 3μM, r‐Moj‐MP and r‐Moj‐DM inhibited 79.67% (+/− 2.88) and 72% (+/− 2.00) cell migration, respectively. The cell migration inhibition between these peptides and the r‐Moj‐WN was significant (p=0.002). At 3μM both echistatin (the positive control) and r‐Moj‐DM strongly inhibited cell migration of HT‐144 cells, 92.59% (+/−6.59) and 93.29% (+/− 4.36), p=0.017 and p=0.007, respectively. Our future studies include the treatment of other cancer cell lines with the r‐Moj‐DM and r‐Moj‐MP peptides in order to determine their effects on cell migration.
An In Vitro Model of HIPEC
Authors: Strom David Kelly , Hoy Samantha D. , Huffman Ryan , Stencel Mike , Boettcher Jessica L. , Hawkins Aruna A. , Larsen Bryan
Published: June 2021

Hyperthermic Intraperitoneal Chemotherapy (HIPEC) is a procedure that utilizes the combination of heat and chemotherapy to kill malignant cells of peritoneal carcinomas that may not be completely removed during cytoreductive surgery. HIPEC has been shown clinically to significantly increase the survival of patients receiving this therapy. In an effort to understand the biological basis of this procedure, we have developed an in vitro model of HIPEC using HeLa cells. HeLa cells show no deleterious effects from incubation at 41°C for 100 minutes. In response to chemotherapeutic agents, HeLa cells are significantly more sensitive to doxorubicin at 41°C as compared to 37°C. However, there was no difference in dose response to Mitomycin C at 41°C compared to 37°C. We also see a 2–4 fold increase in HSP90 protein levels when HeLa cells are incubated at 41°C for 100 minutes. Since Heat Shock Protein expression would likely dampen the killing effects of chemotherapeutic agents, we plan on looking at the expression of both HSP70 and HSP90 in this system, and then use specific HSP blockers to determine if we can potentiate the effects of the chemotherapeutic agents at 41°C. Our goal is to help understand the biological basis of HIPEC and to identify ways to make it even more effective.
Safety evaluation of MJ33 as a potential in vivo inhibitor of NADPH oxidase (NOX2) activation
Authors: Lee Intae , Zagorski John , Dodia Chandra , Feinstein Sheldon , Fisher Aron B.
Published: June 2021

Inhibition of NOX2 has been proposed for the treatment of acute lung injury (ALI) by preventing the generation of reactive O2 species (ROS). MJ33, a fluorinated phospholipid analogue, functions as a potential ALI therapeutic agent by inhibiting the phospholipase A2 (PLA2) activity that is required for NOX2 activation (J. Biol. Chem., 286:11696, 2011). Long‐term toxicity studies were performed to evaluate the feasibility for this use of MJ33. MJ33 incorporated into mixed unilamellar liposomes was injected either intra‐tracheally (IT) or intravenously (IV) in single or multiple doses, up to 25μmol for a 20g mouse. Mice treated with MJ33 up to 10μmol did not show any obvious changes in clinical score, general appearance, activity level, hair quality, body temperature or feces. At 5μmol by IT or IV route, weight loss was <10% and mice recovered with a subsequent small weight gain. Hematocrit and lung wet to dry weight ratio were not altered by MJ33. Weight loss was greater with 10μmol (in multiple doses: 4× 2.5μmol) but was not progressive. Thus, the maximal tolerated dose (defined as causing >15% reduction in body weight) was 0.42μmol/g body wt (~2000 times higher than the dose shown to inhibit PLA2 activity). LD50 was 1.3μmol/g body wt. These results indicate a high margin of safety for administration of MJ33 in the intact mouse.

Supported by NHLBI R01 HL075587
Rimcazole Attenuates the Cocaine‐Induced Stimulation of Mesolimbic Dopamine related to its Abuse and Dependence
Authors: Mereu Maddalena , Chun Lauren E , Hiranita Takato , Cao Jian Jing , Newman Amy , Katz Jonathan L , Tanda Gianluigi
Published: June 2021

Rimcazole (RIM) is a sigma receptor (σR) antagonist that also has affinity for the dopamine (DA) transporter (DAT). Despite this DAT affinity, RIM lacks stimulant effects and dose‐dependently attenuates ambulatory effects of cocaine (COC). RIM and its analogs decrease COC self‐administration in rats (JPET 339: 662), an effect that is obtained neither with selective σR antagonists nor standard DAT blockers.

Thus, we tested RIM (1–17 mg/kg i.p.) alone and in combination with increasing doses of COC that maintain self‐administration (0.1–1.0 mg/kg i.v.), on stimulation of mesolimbic DA in Sprague Dawley rats implanted with a microdialysis probes in the accumbens shell.

RIM dose‐dependently attenuated COC‐induced stimulation of DA levels at doses that had no effects on DA levels when administered alone. Previous studies suggest that the attenuation of COC self‐administration produced by RIM is due to the combined actions at σRs and the DAT as combinations of selective DAT inhibitors and selective σR produced effects like those or RIM.

Thus, the present study indicates that not only can combined DAT and σR inhibition decrease cocaine self administration, it also can decrease its neurochemical substrates suggesting that combined actions at these two targets may serve as a new strategy for discovery of novel compounds for development as medications for stimulant abuse.
The discriminative stimulus effects of bretazenil and lorazepam in rats trained to discriminate pentobarbital from saline
Authors: Hulin Mary Worrel , Winsauer Peter J.
Published: June 2021

Low‐efficacy positive modulators of the GABAA receptor, such as bretazenil, could potentially treat anxiety disorders with less liability for abuse and dependence than traditional benzodiazepines. For this reason, bretazenil was compared with pentobarbital and lorazepam (high‐efficacy positive modulators) in rats trained to discriminate 10 mg/kg of pentobarbital from saline under a fixed‐ratio 20 schedule of food reinforcement. To establish a dose‐response curve in a single session, a cumulative‐dosing procedure was used in which subjects received increasing doses prior to each of four test components. Full substitution was defined as greater than 80% of total lever presses occurring on the pentobarbital‐associated lever. When increasing doses of pentobarbital were administered, pentobarbital‐lever responding increased up to doses that decreased response rate. Unlike pentobarbital, lorazepam only substituted for pentobarbital at doses that decreased response rate. Bretazenil fully substituted in each subject tested. Furthermore, ineffective doses of bretazenil were able to shift the lorazepam dose‐response curve upward and to the left. Together, these data indicate that bretazenil produces pentobarbital‐like discriminative stimulus effects and enhances the effects of lorazepam, indicating that this combination could be clinically useful for long‐term treatment of anxiety disorders.

This work supported by an NIH grant from the National Institute on Alcohol Abuse and Alcoholism AA019848‐01
Elucidating the neurotransmitter systems underlying the subanesthetic intoxicating effects of isoflurane vapor
Authors: Shelton Keith Larry , Slavova-Hernandez Galina , Nicholson Katherine L
Published: June 2021

The present study utilized the drug discrimination paradigm to explore the neurotransmitter systems underlying isoflurane intoxication. Sixteen B6SJLF1/J mice were trained to discriminate 10 min of exposure to 6000 ppm isoflurane vapor from air using a standard 2‐lever operant task. The discriminative stimulus of the benzodiazepine midazolam and the barbiturate methohexital substantially overlapped with that of isoflurane. However, neither the benzodiazepine antagonist flumazenil nor the negative allosteric modulator Ro15‐4515 attenuated isoflurane's discriminative stimulus. The GABAA neurosteroid‐site positive modulator allopregnanolone and the extrasynaptic GABAA receptor agonist, gaboxadol, failed to substitute for isoflurane. Ethanol and the noncompetitive NMDA antagonist, ketamine partially substituted for isoflurane. These findings suggest that the discriminative stimulus effects of isoflurane are the result of GABAA positive modulation and to a lesser extent NMDA antagonism. However, they do not appear to be the result of a direct interaction with the benzodiazepine binding site on the GABAA receptor. Supported by NIDA grant RO1‐DA020553.
Benzodiazepine Self‐Administration in Rhesus Monkeys: Role of α1 subunit‐containing GABAA receptors
Authors: Fischer Bradford D , Platt Donna M , Rallapalli Sundari K , Namjoshi Ojas A , Cook James M , Rowlett James K
Published: June 2021

Benzodiazepines are effective anxiolytics; however drugs in this class also exhibit reinforcing properties which are thought to contribute to their abuse. Recent experimental evidence suggests that benzodiazepine‐induced anxiolytic and reinforcing effects may be mediated by distinct GABAA receptor subtypes. The present study examined the role of α1‐ and α5‐subunit‐containing GABAA receptors in the reinforcing effects of the conventional benzodiazepine triazolam. In rhesus monkeys trained under a progressive ratio schedule of i.v. drug injection, triazolam (0.0001–0.1 mg/kg, i.v.) was reliably self‐administered; resulting in an inverted “U” shaped dose‐response function. Pretreatment with the nonselective antagonist flumazenil (0.01–0.3 mg/kg, i.v.) shifted the triazolam dose‐response function rightward. Similar results were observed with the α1 GABAA receptor‐preferring antagonists βCCT (0.1–1 mg/kg, i.v.) and 3‐PBC (0.3–3.0 mg/kg, i.v.). Schild analysis suggests that each compound antagonized triazolam's effects via a single binding site population. In contrast, pretreatment with the α5 GABAA receptor‐preferring antagonist XLI‐093 (0.003–0.1 mg/kg, i.v.) did not shift the triazolam dose‐response function. Together, these data suggest a role for the α1‐containing, but not the a5‐containing GABAA receptor subtype in the reinforcing effects of benzodiazepines.
Pain relief activates the mesolimbic dopamine reward pathway
Authors: Navratilova Edita , Xie Jennifer , Okun Alec , Eyde Nathan , Ci Shuang , Ossipov Michael , Deeny Tamara King , Porreca Frank
Published: June 2021

Pain is often insufficiently treated by currently available therapies. A major limitation in preclinical models is capturing affective dimensions of pain. We hypothesized that relief of pain would eliminate aversiveness and elicit reward that could be measured using conditioned place preference resulting from negative reinforcement. Additionally, we asked if relief of pain would activate the brain mesolimbic reward circuitry including dopaminergic neurons in the ventral tegmental area (VTA). 24h following incision of the hindpaw, or sham surgery, rats received peripheral nerve block with lidocaine or saline and were paired with a context. Place preference was observed selectively in animals with incision and peripheral nerve block. Additional rats were perfused 2h after lidocaine or saline treatment and processed for immunohistochemistry. An increased number of cFos positive cells was observed within the VTA of rats with incision and peripheral nerve block. Many of these cFos positive cells were also positive for tyrosine hydroxylase suggesting that pain relief activates the reward pathway. Understanding of brain circuits mediating the aversiveness of pain may allow the development of improved therapies. Supported by NS066958‐01.
Phosphorylated serine and threonine residues of PAR2 C‐terminus regulate Beta arrestin 1/2 recruitment and signaling
Authors: Pal Kasturi , DeFea Kathryn
Published: June 2021

Protease activated receptor 2 (PAR2) is activated by tryptic cleavage of the N terminus, to trigger signaling by the classic Gq pathway. Following receptor activation scaffolding molecules Beta arrestin 1/2 bind to C terminus of the receptor to terminate G protein signaling and promote internalization. Simultaneously, this triggers Beta arrestin 1/2 dependent effects like chemotaxis through membrane ERK1/2 activation and cofilin dephosphorylation and PI3kinase activation. The exact role of PAR2 C‐tail phosphorylation in arrestin recruitment and the subsequent downstream effects remain unclear. Bioluminescence Resonance Energy Transfer (BRET) assays using inhibitors of the Gq pathway showed that arrestin recruitment to PAR2 can take place independently of initial Gq signaling. Using PAR2WT, PAR2S363/6A (Protein kinase C mutant) and PAR20P (phosphorylation null mutant) we observed that, Beta arrestin 1/2 recruitment rates are significantly attenuated in absence of PKC mediated C‐tail phosphorylation. Beta arrestin 1/2 recruitment to PAR20P was completely abolished. While PAR2WT promote robust cofilin activation and chemotaxis, PAR2S363/6A and PAR20P fail to do so. Thus, although Beta arrestin 1/2 recruitment to PAR2 C tail can take place in absence of initial Gq signaling, the C‐tail needs to be phosphorylated, to ensure stable association of Beta arrestin 1/2 with PAR2 C‐tails.
Cardiac Scaffold Protein Binding Kinetics: A Role For Altered Signaling During Human Heart Disease
Authors: Rababah Abeer , Craft John , Guillory Ashley , Wijaya Cori , Singh Sonal , Fan Qiying , Diaz Andrea Diaz , Yin Xing , McConnell Bradley
Published: June 2021

Cardiac contractility is physiologically modulated primarily through beta‐adrenergic (β‐AR) receptor signaling. Beta‐adrenergic stimulation signals through phosphorylation of downstream proteins such as protein kinase A (PKA). PKA substrate phosphorylation is facilitated through its co‐localization with its signaling partner by A‐kinase anchoring proteins (AKAPs). In particular, mAKAP (muscle‐selective AKAP) localizes PKA and its target substrates such as phosphodiesterase‐4D3 (PDE4D3). From different genomic databases, we have recently identified fifteen human mAKAP coding non‐synonymous polymorphisms located within or near key protein binding sites critical to β‐AR receptors signaling. Seven of these mutants were cloned for the purpose of comparing whether those substitutions disrupt mAKAP binding to either the PKA binding domain or the PDE4D3. Using surface plasmon resonance (Biacore 2000) we demonstrate specific binding of wild type mAKAP to PDE4D3. Experiments were run in triplicate and as twofold serial dilutions to explore the kinetics of the interaction and analyzed using Scrubber2 with a 1:1 Langmuir model. Comparative analysis of the binding responses of mutations to mAKAP could provide important information about how these mutations modulate signaling.
Kinase activity of phosphoinositide 3‐kinase gamma is needed for T cell development, activation and chemotaxis
Authors: Ladygina Nadia , Fung-Leung Wai-Ping
Published: June 2021

Phosphoinositide 3‐kinase gamma (PI3Kγ) catalyzes the production of phosphatidylinositol‐3,4,5‐triphosphate by phosphorylating phosphatidylinositol (PI), phosphatidylinositol‐4‐ phosphate (PIP) and phosphatidylinositol‐4,5‐bisphosphate (PIP2). Catalytic subunit of PI3Kγ is activated by βγ subunits of heterotrimeric G proteins in response to chemokines, growth factors and hormones, which in turn coordinates cell migration, cell growth, cell cycle entry, and cell survival. PI3Kγ has been implicated in T cell function, but direct effect of its kinase function has not been evaluated. Using mice expressing a catalytically inactive mutant of PI3Kγ(PI3KγKD/KD) we were able to demonstrate that defect in kinase activity leads to T cell developmental defect in the thymus and results in decreased peripheral T cell numbers. PI3KγKD/KD peripheral CD4 and CD8 T cells demonstrated impaired TCR/CD28‐induced proliferation and IL‐2 production, reduced chemokine driven chemotaxis and impaired T cell differentiation. Thus, our study demonstrated that kinase activity of PI3Kγ amend status of T cell response by contributing to thymic development, T cell activation and differentiation as well as lymphocyte chemotaxis.
L‐arginine recycling and endothelial vasomotor function
Authors: Chennupati Ramesh , Meens Merlijn , Janssen Ben , Lamers Wouter , De Mey Jo , Koehler S. Eleonore
Published: June 2021

Objective

To assess whether impaired L‐arginine resynthesis or degradation affects endothelial vasomotor function.

Methods

Endothelium‐selective argininosuccinate synthetase‐ (ASS‐KO) and arginase1‐deficient (Arg1‐KO) mice were generated by crossing ASSfl/fl and Arg1fl/fl mice with Tie2Cre mice. Mean arterial blood pressure (MAP) was recorded in conscious 34‐ week‐old male mice using intra‐arterial catheter. Vasomotor responses were studied in saphenous arteries of 12‐ and 34‐weekold KO and floxed control animals (“WT”) by wire‐myography.

Results

Basal MAP was similar in WT and both KO strains. MAP was increased to the same extent in all genotypes by depletion of circulating L‐arginine (200 U arginase1 intravenously) or inhibition of NOS activity (L‐NAME, 10mg/kg). Optimal arterial diameter, contractile responses to phenylephrine, and relaxing responses to acetylcholine and sodium nitroprusside were also similar in WT and KO mice.

Conclusion

Absence of endothelial arginine recycling or arginine degradation from citrulline does not affect blood pressure and vasomotor responses in small arteries.

Supported by NHF‐grant 2008B107
Brain‐derived Neurotrophic Factor Mediates the Neuro‐protective Effects of Estrone after Brain Injury
Authors: Gatson Joshua W. , Simpkins James , Wigginton Jane , Liu Ming-Mei , Wolf Steven , Minei Joseph
Published: June 2021

Introduction

Estrone (predominant estrogen in postmenopausal women) has shown to be a promising neuro‐protective agent in stroke and other traumatic injuries. The overall goal of this project was to determine if ERK signaling and brain‐derived neurotrophic factor (BDNF) mediates the protective effects of estrone after TBI.

Methods

In this study, male rats were treated with either placebo (corn oil; n=8) or estrone (0.5mg/kg; n=8) at 30 minutes after TBI. Injury was induced using the controlled cortical impact device. Control (n=8) and sham (craniotomy only; n=8) groups were also included in this study. At 72 hours following injury, the brains were fixed and stained for TUNEL + cells. To test for phospho‐ ERK, phospho‐CREB, and BDNF levels, total protein was isolated from brain tissue and Western analysis was used.

Results

Estrone significantly decreased the levels of TUNEL + staining (p<0.01). In addition, in the estrone treated groups there was a significant increase in the levels of phospho‐ERK (p<0.02) and CREB (p<0.05). Also, the levels of BDNF (p<0.007) were significantly increased in the estrone treated groups at 72 hours after injury.

Conclusion

Estrone given after injury is protective and results in an increase in ERK signaling and subsequent BDNF expression. These results suggest that other estrogens in addition to estradiol may afford individuals protection from secondary brain injury after TBI.