bluuebunny/crossref_metadata_2025 huggingface dataset:
PRE‐NATAL LOW PROTEIN DIET IN RATS EVOKES SEX‐DEPENDENT HYPERTENSION IN OFFSPRING
Authors: Augustyniak Robert A. , Singh Karan , Singh Melissa , Rossi Noreen F.
Published: June 2021

Low birth weight is correlated with the development of hypertension later in life. A rat model that is used to study this relationship is reduced protein intake during gestation, which leads to hypertension in the offspring. Previous studies have used imprecise methods to measure blood pressure, such as tail‐cuff. To determine the effect of reduced protein intake during gestation upon blood pressure, 4 day pregnant Wistar dams were placed on an iso‐caloric 6% (low) or 18% (normal) protein diet until term. Male and female offspring (n=6‐8 rats/group) were implanted with a radiotelemetry device at 8 weeks of age to continuously monitor blood pressure in conscious, freely moving rats. At 10 weeks of age, systolic arterial pressure (SAP) was similar between both male groups (118±3 vs. 122±2 mmHg, 6% offspring vs. 18% offspring, respectively, P=NS). However, SAP was significantly elevated in the 6% female offspring compared with the 18% female offspring (117±2 vs. 106±3 mmHg, respectively, P<0.05). Thus, low protein during gestation evokes sex‐dependent hypertension in the offspring. That previous studies have reported hypertension in both sexes of low protein offspring suggest that these rats may have altered responses to stress, which accompanies tail‐cuff blood pressure measurements. Future studies are warranted to determine if low protein offspring have alterations in reflex control of the cardiovascular system.
Estimating Circadian Sleep/Wake Cycle From Rest/Activity Behaviors In Inbred Strains Of Rats
Authors: Feroah Thom R , Vernon H J , McQuestion G , Hutchins W , Forester H V , Ferri R , Manconi M , Merritt A , Dwinell M , Moreno‐Quinn C , Cowley A , Jacob H
Published: June 2021

To establish methods to screen inbred rat strains, we studied EEG/EMG and movement in Brown Norway (BNMCWi), Fawn Hooded (FHMCWi), and Dahl Salt Sensitive (SSMCWi) inbred rat strains in environmental chambers during 3 successive 12L:12D circadian light:dark cycles. The 3rd light:dark cycle was analyzed for the circadian relationship of the onset, offset and duration of sleep state and movement. Although circadian amplitude and phase onset of sleep was different* between strains, onset, offset and duration of sleep and movement were significantly* correlated within strains. Analyzing probability of sleep onset and sleep offset from duration of inactivity we established a 95% probability* of sleep after 35 sec and 0.75 sec, respectively, that was NOT different* within and between strains. We developed an algorithm to determine sleep/wake periods from movement in previously unstudied FHMCWi and BNMCWi rats and found we correctly* estimated sleep onset (90%) and sleep offset (96%) in all observations. We have revised the algorithm to estimate 4 periods of sleep/wake (Active Wake, Quiet Wake, Sleep and Sustained Sleep) to further characterize sleep/wake. In conclusion we have established methods to screen sleep/wake patterns in consomic, congenic and ENU mutated inbred rat strains and to investigate physiological dysfunction and sleep in these models. This study was funded by NHLBI (U01 HL66579) (*P < 0.05)
Estrous cyclicity and the locomotor response to acute and repeated cocaine administration.
Authors: Lopez‐Ojeda Wilfredo , Welsh Matthew L. , Segarra Annabell C.
Published: June 2021

Several studies have shown that gonadal hormones modulate the behavioral response to cocaine. In this study we examined if endogenous changes in sex steroids throughout the estrous cycle affect the locomotor response to repeated cocaine administration. Adult female Sprague Dawley rats were cycled for 21 days. Rats then received a daily injection of saline or cocaine (15 mg/kg,i.p.) for 5 days, followed by 7 drug free days. A challenge injection of saline or cocaine (15 mg/kg,i.p.) was given on day 13. Behavioral activity was recorded on days 1, 5 and 13. Vaginal lavages were performed daily, and on days 1, 5 and 13, after behavioral testing.

Cocaine increased horizontal activity and total distance traveled of drug naive female rats independent of the stage of the estrous cycle. A similar pattern was observed on day 5 of cocaine treatment. However, after 1 week of withdrawal, rats that were in estrous showed higher cocaine‐induced hyperactivity compared to females in proestrous or diestrous. If we compare cocaine‐induced hyperactivity of females at different stages of the estrous cycle we observe that only females in estrous show an increase from day 1 to day 13, indicating that females in estrous become sensitized to cocaine. These results are similar to those obtained by other groups. An unexpected finding was the shortening of the estrous cycle. In summary, we observed that the stage of the estrous cycle affects the locomotor response to repeated cocaine administration. These data may have implications in treatment strategies for female cocaine addicts.
Exercise‐induced changes in the Na+,K+‐ATPase Na+ affinity in rat skeletal muscle
Authors: Juel Carsten , Walas Helle
Published: June 2021

Regulation of the Na+,Kb‐ATPase in skeletal muscle is important for ion homeostasis and muscle function. We have used the inorganic phosphate release to quantify the Na+‐induced Na+, K+‐ATPase activity in purified membranes from oxidative and glycolytic rat muscle. In membranes purified from control rats the Km for Na+ was lower in oxidative compared to glycolytic fibers (6.1 vs. 14.0 mM) (P<0.05). In membranes purified after 30 min of treadmill running the Km for Na+ was increased (P<0.05) to 15.8 mM in oxidative fibers, whereas Km for Na+ (11.6 mM) in glycolytic fibers was not different from control. Analysis of subunit (α, β and FXYD1) protein expression in the sample used revealed fiber type dependent differences in subunit content, a moderate activity‐dependent increase in α and βsubunits (translocation), and a two‐fold increase in the FXYD1(PLM) content. It is concluded that the affinity for Na+ is higher in oxidative compared to glycolytic fibers, and that muscle activity reduces the Na+ affinity in oxidative fibers. The underlying mechanisms may include translocation of subunits to the membrane fraction studied, phosphorylation of subunits, and increased association of the FXYD1 subunit with the αβ‐complex. The affinity differences may explain the fiber type dependent differences in ion changes associated with muscle activity.
Up‐regulation of enzymes involved in reactive oxygen species (ROS) metabolism in skeletal muscle of mice lacking neuronal nitric oxide synthase (nNOS)
Authors: SilvaAzevedo Luis Da , Baum Oliver , Jaehne Sebastian , Hoffmann Christian , Stalder Daniel , Heller Manfred , Zakrzewicz Andreas , Pries Axel
Published: June 2021

nNOS knockout mice do not show any phenotypic malfunctions, thus we hypothesized that cellular mechanisms exist which compensate for nNOS deficiency. In order to identify molecular compensatory mechanisms as well as to gain new insights into the function of nNOS in skeletal muscle, we searched for proteins differentially expressed in extensor digitorum longus muscle (EDL) of nNOS knockout mice compared to C57/Bl6 wild type mice, by two‐dimensional polyacrylamide electrophoresis (2D‐PAGE) and immunoblot. About 800 spots were detected on silver‐stained gels after densitometric analysis. Analysis of silver‐stained gels showed one protein with Mr of 24.5 kDa and pI of 5.9 present in EDL of nNOS−/− mice but not of C57 mice. This protein was identified as peroxiredoxin‐6 (PRX6) by LC‐MS/MS and immunoblot. Five other proteins were found to be present in higher amounts in EDL of nNOS‐knockout mice: peroxiredoxin‐3 (PRX3), prohibitin, superoxide dismutase, heat shock protein beta‐1 and nucleoside diphosphate kinase A. Up‐regulation of PRX6, PRX3 and heat shock protein beta‐1 was also observed at the mRNA level. The in vitro production rate of total ROS was significantly higher in EDL of nNOS knockout mice. Furthermore, treatment of C57 mice with the NOS inhibitor L‐NAME induced PRX6 in EDL. Induction of PRX6 in nNOS−/− mice could have a protective role in regulating levels of H202, 02‐ as well as oxidised phosholipids.
Comparability testing of KRAS mutations and association with efficacy from a phase 3 randomized trial of panitumumab (pmab)
Authors: Patterson Scott D , Oliner Kelly , Freeman Daniel , Juan Todd , Wolf Michael , Sarosi Ildiko , Bakker Andreas , Van Cutsem Eric , Peeters Marc , Siena Salvatore , Reese David , Chang David
Published: June 2021

Background

KRAS mutation status is an important predictor of response to epidermal growth factor receptor inhibitors.

Methods

KRAS mutational status was tested on 40 colorectal cancer (CRC) samples (obtained through Asterand, Ltd. [USA]) in 4 independent commercial laboratories and by direct sequencing at Amgen. Kappa statistics (κ) were computed to compare the commercial test results to the sequencing results. Following this evaluation, HistoGeneX tested tumor samples from a randomized phase 3 trial of pmab in chemotherapy‐refractory metastatic CRC.

Results

Allele‐specific tests resulted in the highest correlation, κ=0.94 (Genzyme) and 0.90 (HistoGeneX), to the Amgen sequencing results. The direct sequencing Gentris test yielded κ=0.75. For these 3 tests, p<0.0001. Oligonucleotide hybridization (Invitek) yielded κ=0.13, p=0.3324. In the phase 3 trial, 184/427 (43%) of pts had tumors with mutant (MT) KRAS. Partial responses and stable disease, respectively, were observed in 17% and 46% of pts without MT KRAS vs 0% and 20% of pts with MT KRAS.

Conclusions

Allele‐specific PCR‐based methods from Genzyme and HistoGeneX were comparable with the Amgen direct sequencing method and may be utilized as accurate, readily‐available tests for KRAS mutations. The DxS TheraScreen KRAS kit, employed by HistoGeneX, has also demonstrated utility in clinical trials.
The effect of diet and polybrominated diphenyl ether exposure on adipocyte and whole body metabolism in male Wistar rats
Authors: Allgood Erin Leigh , Carey Gale
Published: June 2021

Polybrominated diphenyl ethers (PBDEs) are lipophilic flame‐retardant chemicals found in many consumer products and increasingly in human tissues. The purpose of this study was to elucidate whether exposure to PBDEs in conjunction with a high‐fat/high‐sugar (HF) diet will predispose male Wistar rats to obesity. Twenty four rats were fed either a control (C) or HF diet, and gavaged with either 18 mg/kg PBDEs (+) or corn oil (‐) daily for 4 weeks (n=6 per group). Body weight and food intake were measured three times per week. At 3 weeks, 24‐hr whole body metabolism was measured. At 4 weeks, blood was sampled for plasma thyroid hormone concentration (T4), epididymal fat pads were removed and weighed, and adipocytes were isolated. Adipocyte size and lipolytic response to adenosine deaminase and varying concentrations of isoproterenol over 90 min were measured. Findings revealed that there were no significant differences in lipolysis with ADA or isoproterenol. A slight but significant increase in % body weight change was seen in + vs. ‐ rats. HF rats consumed 9% more kcals than C rats over the 4 week study (P<0.05). T4 levels decreased by 83% in + vs. ‐ rats (P<0.05). Whole body metabolism results are pending. PBDEs cause an increase in body weight and reduction in T4 without affecting adipocyte size and lipolytic sensitivity; the mechanism by which this occurs is unknown. Supported by Agric. Expt. Stn. Grant NH 00485.
Mirthful laughter, as adjunct therapy in diabetic care, increases HDL cholesterol and attenuates inflammatory cytokines and C‐RP and possible CVD risk
Authors: Berk Lee S , Tan Stanley
Published: June 2021

Purpose

Catecholamines are implicated in causing arrhythmias, hypertension, and MI reoccurrence. Mirthful laughter attenuates catecholamines. Inflammatory cytokines, TNF‐[alpha], IFN‐[gamma], and IL‐6 can accelerate atherosclerotic disease. Hs‐CRP is a risk marker for the inflammatory process and CVD. Mirthful laughter, by diminishing catecholamine production, can attenuate inflammatory cytokines and provide CVD protection.

Methods

Twenty high risk diabetic patients with hypertension and hyperlipidemia were age and sex matched and divided into Group C (control) and Group L (laughter). Group C received standard medication therapies for diabetes, hypertension, and hyperlipidemia. Group L, additionally, viewed self‐selected humor 30 minutes daily. Blood was assessed at baseline and every 2 months, for 12 months, for plasma HDL‐cholesterol and inflammatory cytokines TNF‐[alpha], IFN‐[gamma], IL‐6, and hs‐CRP.

Results

Compared to Group C, the patients in Group L had by the 2nd month increased HDL cholesterol and by the 4th month decreased TNF‐[alpha], IFN‐[gamma], IL‐6, and hs‐CRP blood levels (p<0.01).

Conclusions

The addition of mirthful laughter as a preventive adjunct therapy in diabetes care raises HDL cholesterol levels and lowers inflammatory cytokines, and hs‐CRP levels. These modulation effects can contribute to lower CVD risk and fewer MI reoccurrence.
The influence of empirically derived dietary patterns on cognitive function in independent older adults depends on income and education: The NuAge Study
Authors: Parrott Matthew , Shatenstein B , Ferland G , Morais JA , Belleville S , Kergoat MJ , Payette H , Greenwood CE
Published: June 2021

This study prospectively examined the relationships between global cognitive function (Modified Mini‐Mental State examination; 3MS) and dietary patterns derived from a semi‐quantitative food frequency questionnaire by principal components analysis. Relationships were examined using linear mixed models in 1266 independent senior citizens assessed annually over 4 years. After adjustment for health and socioeconomic variables, higher adherence to a ‘prudent’ diet pattern associated with better overall 3MS performance (P=0.046) while greater adherence to a ‘Western’ diet pattern associated with poorer overall 3MS performance (P=0.018). Nevertheless, diet patterns interacted with either household income (P<0.01) or education (P<0.01) in fully adjusted models. Specifically, income was related to rate of cognitive decline only in participants with below median prudent pattern score (P=0.015) such that higher income protected against 3MS decline from baseline in this group. The negative impact of Western pattern score on overall 3MS performance was apparent in participants with below median education (P<0.01), but absent in those with above median education. These results are consistent with cognitive reserve theory, and its proposed impact on cognitive performance in the elderly, suggesting that the influence of diet on cognition should be considered in this context. (CIHR)

Grant Funding Source: CIHR
African American perceptions of heart disease and nutrition in Arizona
Authors: Winham Donna M , der Ananian Cheryl , Thompson Sharon V
Published: June 2021

African Americans have the highest rates of cardiovascular disease in Arizona. The project identified perceptions and knowledge of heart disease among African Americans aged 25–60 years through 15 focus groups (FG). A total of 104 men (44%) and women (56%) completed the focus groups and questionnaires on dietary intakes, behavioral risk factors, neighborhood characteristics, and ethnic identity. Analysis revealed the following qualitative themes: 1) recognition of the role of diet in heart disease etiology; 2) acknowledgment of poor eating habits due to lack of time and money; 3) inability to make traditional foods healthy; 4) healthy foods were readily available in their neighborhoods; 5) feelings of social isolation because African Americans are dispersed over metro Phoenix area. In terms of risk factors, the majority of the sample was overweight or obese. The prevalence of smoking was significantly higher in men (58%) than in women (26%) (p.000). Fifty‐one percent of the men exceeded dietary recommendations for fat intake in comparison to 30% of the women. Few people reported eating Soul Foods regularly, except for greens or fried fish, which were consumed at least 2–3 times per month by 50% of the sample. These findings suggest points to improve health education based on existing knowledge and perceptions. Funding was provided by the Arizona Department of Health Services.
Functional Role of Xanthophyll Metabolism Related Genes in Human Retinal Pigment Epithelial Cells
Authors: Gong Xiaoming , Marisiddaiah Raju , Afeld Jamiee , Wiener Doris , Rubin Lewis P.
Published: June 2021

Xanthophyll carotenoids (lutein, zeaxanthin) preferentially accumulate in the macula of the human retina. They play a critical role in protecting the macula from oxidant and photo‐damage. Previous studies have indicated macular pigment density (xanthophyll content) is inversely related to adult onset age‐related macular degeneration (AMD). The regulation of macular pigment density in retinal pigment epithelium (RPE) is under active investigation.

We determined the expression of genes related to carotenoid uptake and metabolism in human retinal pigment epithelial (ARPE‐19) cells using message (qRT‐PCR) and protein (Western blot). Xanthophyll uptake and transport were assayed using polarized ARPE cells grown to confluence.

ARPE cells express scavenger receptors (SR‐B1, CD36, LDLR but not SR‐A), carotenoid cleavage enzymes (CMO1, CMO2) and transport related protein (ABCA1). RPE expresses high levels of SR‐B1, LDLR, CMO2 and ABCA1, and lower expression of CD36 and CMO1. RPE exposure to specific carotenoids (beta‐carotene, lycopene, lutein, astaxanthin) induces different expression patterns. Furthermore, gene expression profiles significantly changed when RPE cells were exposed to graded oxidative stress. Finally, we determined that lutein uptake and transport is directional across polarized ARPE monolayers: significantly higher from basolateral to apical surface than in the opposite direction. Of note, co‐incubation with beta‐carotene decreased lutein uptake and transport rates.

These findings support the hypothesis that xanthophyll (especially lutein) uptake, metabolism, and transport are critical in macular pigment homeostasis. We speculate that altering RPE gene expression profiles by protective (xanthophyll) or deleterious (oxidative stress) factors point to molecular mechanisms in the pathogenesis of AMD. (NIH HD42174, Muma Family Endowment)
Dietary Cod Protein Improves Skeletal Muscle Regeneration Partly Through Its High Levels of Arginine, Lysine, Glycine and Taurine
Authors: Dort Junio , Leblanc Nadine , Côté Claude , Jacques Hélène
Published: June 2021

Feeding cod protein may beneficially influence the time course of inflammation during recovery following skeletal muscle injury. The objective of this study was to identify amino acids in cod protein having a beneficial impact on muscle repair. Male Wistar rats were fed isoenergetic diets containing either casein (C), cod protein (CP), or casein supplemented with arginine (1.9%), glycine (1.9%), taurine (0.7%) and lysine (1.7%) (C+). After 21 d of ad libitum feeding, one tibialis anterior muscle (TA) was injured by injecting 200μl of bupivacaine while the contra‐lateral TA was injected with saline and served as sham. Rats fed CP exhibited higher muscle weight at days 5, 14, and 28 post‐injury and higher myofiber cross‐sectional area at days 5 and 28 post‐injury compared with C. CP and C+ similarly modulated the inflammation as they decreased ED1+ macrophages and COX‐2 level at day 2 post injury compared with casein (p=0.01), and increased ED2+ macrophages compared with C. The C+ diet upregulated (p=0.02) whereas the CP diet tended to upregulate myogenin expression at day 5 post‐injury compared with C (p=0.06). These data support the hypothesis that the beneficial impact of cod protein on skeletal muscle recovery is partly attributed to the anti‐inflammatory and myogenic action of its high arginine, lysine, glycine and taurine levels.

Supported by The Natural Sciences and Engineering Research Council of Canada.
The effects of combination treatment of parenteral arginine and a nitric oxide synthase inhibitor L‐NAME on oxidative stress in rats with sub‐acute peritonitis
Authors: Lo Hui-Chen , Lee Chien-Hsing
Published: June 2021

Using a modified rat model with cecal puncture and sub‐acute inflammation, we investigated the combination effects of arginine and L‐NAME, a nitric oxide synthase inhibitor on oxidative stress and antioxidant enzymes in the circulation and peritonitis‐affected target organs. Male Wistar rats with sub‐acute peritonitis were infused with a conventional parenteral nutrition solution or that supplemented with arginine, L‐NAME or both. Parenteral fed rats without peritonitis were used as controls. After 7 days, peritonitis‐induced alterations in TBARS, the lipid peroxidation products, in the erythrocytes and kidneys were significantly reversed by parenteral arginine. In addition, the increased activities of superoxide dismutase (SOD) and catalase in the liver were significantly alleviated by arginine supplementation. L‐NAME significantly decreased the activity of glutathione peroxidase in the liver and significantly increased the levels of vitamin C in the plasma and the activity of SOD in the kidneys. Combination treatment significantly increased plasma vitamin C and pulmonary thiol group and glutathione levels. In conclusion, our results showed that parenteral arginine and L‐NAME may modify antioxidant enzymes and non‐enzyme substrates, respectively. In addition, combination treatment allows the peritonitic rats to maintain an oxidant‐antioxidant homeostasis with non‐significant alterations in lipid peroxidation and antioxidant enzyme activities.
Clearance of glutamine in mares
Authors: Manso Helena Emilia Cavalcanti C.C. , Manso-Filho Hélio Cordeiro , de Oliveira Deyvson Ferreira , de Aquino Rafael Santos , Gouveia-Júnior Carlos Roberto Ribeiro , Nogueira Eduardo Terra , kutschenko Marianne , DˈPaula Joyci Torres , Mélo Stephânia Katurchi , de Almeida Telga Lucena A. C. , Bernardo Ramona Bastos , Watford Malcolm
Published: June 2021

Supplementation with glutamine has been proposed to prevent the catabolic response to exercise but there are few studies of glutamine supplementation in training horses. Four Arabian mares were divided in a latin square design to determine the optimal dose of glutamine. The mares received zero, 10g, 20 and 40 grams of AminoGut (L‐glutamine and L‐glutamate) mixed into one kilogram of the meal. Blood was drawn prior to the meal and at 4h post‐feeding. There were no differences in plasma urea, creatinine and uric acid over 4h. Plasma glutamine rose in the AminoGut supplemented group with a peak at 4h. There were no differences between the group that received 20g or 40g of AminoGut. The results indicate that 20g or 2% of AminoGut in the diet could be beneficial to prevent catabolism during exercising horses.
The Effect of Feeding Cinnamon on Postprandial Blood Glucose and Lactate in Trained Endurance Athletes
Authors: Ongkoputro Ella , Burns-Whitmore B. , Spalding Thomas , Sokmen Bulent , Bidlack Wayne
Published: June 2021

Research studies indicate polyphenol polymers, also found in cinnamon, have insulin‐potentiating properties. Our objective was to determine if six grams of cinnamon significantly reduces postprandial blood glucose and blood lactate in trained male endurance cyclists.

Eleven trained male cyclists participated in the study (28.8 ±6.4 yo; maximal oxygen consumption of 57.9±7.5 ml∙kg‐ 1∙min‐1). Participants were randomly assigned to either a cinnamon (6 g) or control treatment along with a pre‐exercise meal: 76g of wheat bread, 7.5g of unsalted butter, and 6g of sugar. Blood glucose and blood lactate were measured at minute 0, 30, 60, 90 and end of exercise. Subjects finished both treatments.

Results indicate a significant time main effect (p< 0.001); mean blood glucose at Min 0 was greater than before the meal, and mean blood glucose decreased significantly from Min 0 to Min 30 during exercise and remained stable through Min 90. The change pattern in mean blood lactate across time was similar in both conditions, and blood lactate did not rise above pre‐exercise levels during 90 minutes of exercise at 70% of VO2 max, but rose substantially and significantly as a result of exercise at 85% of VO2 max (p< 0.05).

We conclude that ingestion of cinnamon prior to endurance exercise does not exert significant effects in lowering postprandial blood glucose and blood lactate in trained male endurance cyclists.
Lipocalin‐2 Serum Levels Decrease in a Randomized Dietary Feasibility Study for Stage II‐IV Ovarian Cancer Survivors
Authors: Garcia-Prieto Celia , Paxton Raheem , Berglund Maria , Hernandez Mike , Hajek Richard A. , Handy Beverly C. , Brown Jubilee , Jones Lovell A.
Published: June 2021

Background

Lipocalin 2 has been shown to be up‐regulated in ovarian cancers and correlated with disease progression but the role of fruit and vegetable consumption on serum levels in ovarian cancer survivors has not been studied.

Methods

In a randomized design, women diagnosed with stage II‐IV ovarian cancer were assigned to a low fat, high fiber diet or a modified National Cancer Institute diet supplemented with a soy‐based beverage and encapsulated fruit and vegetable juice concentrates (FVJCs). Blood samples were collected at 0, 3, 6 months and serum levels of lipocalin‐2 were determined using highly sensitive immunoassays.

Results

The FVJC dietary intervention significantly lowered lipocalin‐2 serum levels compared to LFHF after 6 months (P<0.05). Those participants in the FVJC group above the average age and with no history of oral contraceptive use experienced the most significant decreases (P<0.05). Correlations between lipocalin‐2 levels with body mass index, or CA‐125 did not indicate significance.

Conclusion

These results show evidence that a low fat, high fiber diet supplemented with fruit and vegetable juice concentrates can cause decreases in lipocalin‐2 serum levels after 6 months of consumption and suggest further studies are necessary to investigate mechanism.
Inhibitory effects of phloretin on thrombin induction of protease activated receptor and inflammatory cytokines
Authors: Kim MinSoo , Kang Young-Hee
Published: June 2021

Beyond its key role in the dynamic process of physiological and pathological coagulation as a proteolytic enzyme, thrombin possesses a distinct pro‐inflammatory activity, which may influence the onset and progression of atherosclerosis. This study investigated with THP‐1 monocytes that 1–20 μM phloretin, a dihydrochalcone found in apple tree leaves, attenuated thrombin induction of protease activated receptor (PAR) and inflammatory cytokines being involved in early atherosclerosis. This study found that thrombin promoted its membrane receptor PAR‐1 expression, which was blunted by phloretin nontoxic at 1–20 μM. The thrombin induction of plasminogen activator inhibitor‐1, a serine protease inhibitor, was dose‐dependently attenuated by treating phloretin to monocytes. In addition, the secretion of the cytokines of monocyte chemotactic protein‐1, interleukin (IL)‐6 and IL‐8, all responsible for vascular inflammation, was enhanced by thrombin. Such secretion was dampened by submicromolar phloretin, as evidenced by Western blotting and enzyme‐linked immunosorbent assay. These results demonstrate that phloretin retarded the expression and secretion of PAR‐1, PAI‐1 and inflammatroy cytokines in thrombin‐experienced monocytes. Thus, phloretin may qualify as a potential agent targeting thrombin exerting pro‐atherogenic actions associated with vascular inflammation.
Effects of prickly pear cactus (Opuntina ficus‐indica var. Saboten) intake on blood lipids, platelet aggregation, antioxidant and liver parameters in volunteer women
Authors: Han Sun-Geum , Kim Mi-Sook , Kang Young-Hee , Kang Jung-Sook
Published: June 2021

We analyzed dietary effects of prickly pear cactus (Opuntina ficus‐indica var. Saboten) on plasma lipids, platelet aggregation, hemolysis, plasma TBARS and liver parameters after daily intake of 20g cactus tea containing 27% prickly pear cactus powder for 4 weeks using 14 normotensive and 14 hypertensive volunteer diving women living in Jeju island, south Korea.

Plasma total cholesterol and HDL cholesterol were not changed in both groups of the normotensive and the hypertensive after cactus intake. Plasma triglyceride was decreased by 20% in the hypertensive and by 50% in the hyperlipidemic subjects (≥150mg/dl, n=9). Platelet aggregation in both maximum and initial slope were significantly decreased only in the normotensive or subject with normal cholesterol (<200mg/dl, n=11) after cactus intake (both, p<0.01). Total bilirubin was significantly decreased (p<0.05) and sGOT was significantly increased (p<0.05) in all categories (n=28) after cactus intake. Alkaline phosphatase was decreased significantly only in the hypertensive subjects after intake (p<0.05, n=14). Hemolysis after 6 hours isotonic incubation was significantly decreased in all categories after intake (p<0.01, n=28). TBARS production was significantly reduced only in the hypercholesterolemic (p<0.01, n=17) or in the hyperlipidemic (p<0.05, n=9) cases after cactus intake.

Present study showed that flavonoids and soluble fiber rich prickly pear cactus may have hypolipidemic, antithrombotic, antioxidant effects differently depending on the subject conditions in term of blood pressure, plasma lipids and others cardiovascular related.
Effect of Coenzyme Q10 and Ardisia japonica Blume (AJB) on plasma and liver lipids, platelet aggregation and RBC Na efflux channels in simvastatin treated hypercholesterolemic guinea pigs
Authors: Kang Min-Sook , Yang Hun-Mo , Kang Ja-Young , Kang Jung-Sook
Published: June 2021

Forty guinea pigs were divided into four groups and fed 0.5% cholesterol based control diet, plus 0.05% simvastatin, statin plus 0.15% CoQ10 or 10% Ardisia japonica (AJB) leaves for 4 weeks. Plasma total cholesterol was significantly decreased in all groups fed statin containing diets compared with the control diet (p<0.01). Plasma and liver triglycerides were significantly decreased in group with statin plus CoQ10 compared with the control (both p<0.05). Maximum platelet aggregation was significantly higher in group with statin plus CoQ10 than other groups (p<0.05). Na‐K ATPase was increased in group with statin and decreased in group with statin plus CoQ10 and the difference between these two was significant (p<0.01). Na‐K cotransport and Na passive transport were significantly decreased in the control group compared with other groups (both p<0.05). Intracellular Na was the highest in group of statin and the lowest in group of statin plus CoQ10, correlating with Na‐K ATPase. TBARS productions in platelet rich plasma and liver tended to decrease in the groups with statin plus CoQ10 compared to other groups. sGOT and sGPT were significantly increased in group with statin compared with the control (p<0.05), and statin plus CoQ10 decreased the enzymes to the control levels.

In present study, antioxidant rich AJB did not have positive effects in CVD parameters. Besides the beneficial effects as antioxidant and liver protection, statin plus CoQ10 seemed to decrease cholesterol more efficiently than statin alone.
9‐cis‐Retinoic acid induces integrin‐independent immune cell adhesion
Authors: Whelan Jarrett T. , Chen Jiaming , Morrow Rebekah L. , Miller Jabin , Merrell Kaitlin S. , Lingo Joshuah D. , Shaikh Saame R. , Bridges Lance C.
Published: June 2021

Retinoids are essential in the proper establishment and maintenance of immunity. Although retinoids are implicated in immune related processes, their role in immune cell adhesion has not been well established. In this study, we investigated the adhesive effects of 9‐cis‐retinoic acid (9‐cis‐RA) on a variety of human hematopoietic cell lines. 9‐cis‐RA treatment specifically induced immune cell adhesion of the human B‐cell line RPMI 8866, and the human monocytic cell line U937 to select immune ligands. Due to the prominent role of integrin receptors in mediating immune cell adhesion, we sought to evaluate if 9‐cis‐RA induced RPMI 8866 and U937 cell adhesion was integrin dependent. Intriguingly, 9‐cis‐RA mediated cellular adhesion was not affected by a variety of function blocking integrin inhibitors. In addition, the adhesion of RPMI 8866 cells could be inhibited by treatment with synthetic corticosteroids, suggesting a potential role in the inflammatory response. Finally, we demonstrate that 9‐cis‐ RA treatment of primary murine B‐cells also induces ex vivo adhesion. Our study is the first to demonstrate that 9‐cis‐retinoic acid prompts the cellular adhesion of immune cells, in vitro and ex vivo, through a yet unidentified immune receptor/complex whose expression or activation is induced upon 9‐cis‐RA exposure.
Complementary food choices from 6–18 mo: effect on iron (Fe) status and inflammation
Authors: Sheng Xiao-Yang , Ma Jing-Qiu , Liu Jin-Rong , Hu Yan-Qi , Liu Shan-Shan , Zhang Jie , Westcott Jamie , Hambidge Michael , Krebs Nancy F
Published: June 2021

Objective

To compare Fe status in rural poor 18 mo old toddlers fed 3 different complementary foods.

Methods

The study was a cross‐sectional convenience sub‐sample of a larger study in which 6 mo old infants were randomized to receive 50g/d pork (MG, n=137), an equi‐caloric micronutrient fortified cereal‐based supplement (FG,140) or local non‐fortified rice supplement (LG, 133) for 1 yr. Hb, sTfR, HsCRP, ferritin, AGP were measured in serum; calprotectin (FCP) was measured in feces. Body Fe =‐log (sTfR) × 1000/ferritin)‐2.8229]/0.1207. Fe deficiency=ferritin<12μg/L.

Results

Mean (± SD) Hb, MCV, MCHC, sTfR, ferritin did not differ among groups. Significant differences were:

Parameter
MG
FG
LG
P‐value

Body Fe, mg/kg1
6.6 ± 3.1
7.6 ± 2.6
6.3 ± 3.4
<0.01

Fe deficiency, n (%)1
40 (34%)
26 (25%)
46 (41%)
0.03

AGP, g/L, (n)
0.75±0.21(37)
0.98±0.33(39)
0.83±0.35(31)
<0.01

FCP, μg/g, (n)
200±214(37)
413±491(39)
192±197(31)
<0.001

Excluded subjects with Hs‐CRP>3mg/L

Conclusion

Micronutrient fortified cereal improved Fe status but was also associated with systemic and intestinal inflammation. Supported by Thrasher Research Fund; Science and Technology Commission of Shanghai Municipality; Shanghai Key Laboratory of Children's Environmental Health.
Effect of selected dietary regimens on recovery from moderate acute malnutrition in young Malian children
Authors: Ackatia-Armah Robert S , McDonald Christine , Doumbia Seydou , Brown Kenneth H
Published: June 2021

Moderate acute malnutrition (MAM) remains a major public health concern and research is needed on optimal dietary treatment regimens. We randomly assigned 12 rural health centers in Dioila Health District, Mali to provide 1 of 4 dietary regimens approximating WHO recommended nutrient profiles: 1) lipid‐based, ready‐to‐use supplementary food (Plumpy'Sup, PS); 2) special corn soy blend for MAM (CSB++); 3) locally processed, fortified food (Misola); or 4) locally milled flours and micronutrient powder. 1284 non‐edematous children aged 6–35 mo with initial arm circumference (MUAC) <12.5 and ≥11.0 cm or weight‐for‐length Z‐score (WLZ) <‐2.0 WHO standard and ≥70 % NCHS reference median were offered ~500 kcal/d of assigned diet in addition to home food intake for 12 wks. Among 1121 children (87.3%) who completed the study, the mean±SD change in weight from baseline was greater with PS than all 3 other regimens, which did not differ from each other (1.15±0.57 kg, 0.96±0.55 kg, 0.96±0.53 kg, 0.91±0.54 kg for respective study groups, p<0.001 ANOVA). MUAC changes followed a similar pattern, but length did not differ significantly among groups. Recovery from MAM (WLZ>‐2.0, MUAC>12.5cm) was highest with PS compared with the 3 other regimens (71%, 56%, 59%, 53% for respective groups, p<0.001 Scheffe). PS is more effective than the other dietary regimens, but benefits must be considered in relation to product costs and availability. This study was supported by UNICEF and WFP.

Grant Funding Source: UNICEF and WFP
D295N Mutant Cathepsin D Exerts a Dominant Negative Effect In Vitro by Promoting α‐Synuclein Accumulation
Authors: Crabtree Donna Marlana , Ouyang Xiaosen , Zhang Jianhua
Published: June 2021

Parkinson's disease (PD) is a neurodegenerative movement disorder affecting more than one percent of people over the age of 65. The histopathological hallmark of PD are proteinaceous inclusions termed Lewy bodies, which contain aberrantly accumulated α‐synuclein. Current therapeutic strategies are aimed at reducing α‐synculein burden within the brain, and understanding factors that contribute to early events which cause synuclein accumulation may facilitate this aim. Cathepsin D (CD) is a lysosomal aspartic protease which has been previously demonstrated to cleave α‐synuclein and reduce α‐synuclein‐associated toxicity both in vitro and in vivo. We show here that introduction of a catalytically inactive mutant CD via a lentiviral vector in SH‐SY5Y cells results in inhibition of endogenous wildtype CD activity and accumulation of α‐synuclein. To further explore the mechanism behind mutant CD's effect on α‐synuclein we used PepstatinA (PepA) to pharmacologically inhibit CD activity. We observed that use of PepA also resulted in significantly increased levels of α‐synuclein. Additionally, LC3I levels were significantly increased in both cells expressing mutant CD or cells treated with PepA, suggesting alterations in the autophagy pathway result from inhibition of CD activity. Our results suggests a dominant negative function for the catalytically inactive mutant CD. We demonstrate that decrease of endogenous CD activity in dopaminergic cells gives rise to α‐synuclein accumulation and alterations in the autophagic pathway, suggesting that improving and/or maintaining CD function within the brain may represent a way to prevent α‐synuclein aggregation and corresponding neuropathology from occurring.
Reversible tetracyline‐controlled transactivator (rtTA)‐ inducible expression of neuron‐targeted Cav‐1 and recovery after neuronal injury
Authors: Grove Anna M H , Kellerhals Sarah E , Stary Creed M , Patel Hemal H , Patel Piyush M , Head Brian P
Published: June 2021

Decreased capacity for neuronal regeneration following brain injury or neurodegeneration is due to a growth inhibitory environment resulting from decreased expression of pro‐survival and pro‐growth pathways. Caveolin‐1 (Cav‐1), a protein found in membrane lipid rafts, regulates pro‐survival and pro‐growth signaling and cAMP formation. We have shown that synapsin driven neuron‐targeted Cav‐1 over‐expression (SynCav1) enhances pro‐growth signals, dendritic arborization, and axonal sprouting, even in the presence of growth inhibitors. To test the efficacy of SynCav1 in an injury model, we have generated a reversible tetracycline transactivator (rtTA) neuron‐targeted Cav‐1 vector (Syn‐rtTA‐tetO‐Cav1). Primary neurons transfected with Syn‐rtTA‐tetO‐ Cav1 and 72 hr doxycycline (DOX, 10 ug) treatment increased Cav‐1 expression. Based on such vector data, we have generated two novel transgenic mouse lines, a constitutively active (SynCav1TG) and an inducible (rtTA SynCav‐1rtTA) neuronal Cav‐1. These two transgenic lines will be tested in our traumatic brain injury model (controlled cortical impact). Neuron‐targeted Cav‐1 expression in the setting of injury may be a novel therapeutic control point for limiting spread of injury and neurodegeneration.

This project was funded by VA CDA‐2
Synergistic interaction between scriptaid and proteasome inhibitors enhances the sensitivity of human colorectal cancer cells to chemotherapeutic drugs
Authors: Abaza Mohamed Salah , Bahman Abdul-Majeed , Al-Attiyah Rajaa
Published: June 2021

Histone deacetylase inhibitors (HDACis) represent a promising new class of compounds for the treatment of cancer. Exposure of cancer cells to HDACis leads to activation of nuclear factor kappa B, such finding raise the possibility that co‐administration of HDACis with proteasome inhibitors (PSis) which interrupt this pathway may be associated with enhanced anticancer activity. To address this issue we have examined the effects of combined treatment with HDACi, scriptaid and proteasome inhibitors; the molecular mechanisms of the combined treatment and its potential to sensitize cancer cells to chemotherapeutic drugs.

Inhibition of colorectal cancer cell growth by scriptaid was potentiated by PSis, clastolactocystin, MG132, PSi‐I or epoxomocin in a dose dependent manner. The combined treatment inhibited cell cycle, induced apoptosis and differentially sensitized cancer cells to chemotherapeutic drugs of different mechanisms of action. Co‐administration of scriptaid and PSis is associated with enhanced anti‐proliferative activity, apoptosis and chemo‐sensitization of colorectal cancer cells through a selective induction or suppression of genes that play important roles in cell cycle and apoptosis.

The present study provides evidence that the combination of scriptaid and proteasome inhibitors shows a strong synergistic interaction with chemotherapeutic drugs in colorectal cancer cells, providing the foundation for future clinical translation with HDAC and proteasome inhibitors for anticancer therapy.
Effects of the Melatonin Receptor Antagonist (MT2)/Inverse Agonist (MT1) Luzindole on Re‐entrainment of Wheel Running Activity and Spontaneous Homecage Behaviors in C3H/HeN Mice
Authors: Adamah-Biassi Ekue Bright , Stepien Iwona , Hudson Randall L , Dubocovich Margarita L
Published: June 2021

Activation of MT1 receptors by melatonin accelerates the re‐entrainment of circadian rhythms of wheel running activity after an abrupt advance of the dark cycle (Dubocovich et al., 2005). Here, we investigated the effects of luzindole (LUZ), a competitive melatonin receptor antagonist (MT2)/inverse agonist (MT1) on the re‐entrainment rate of wheel running activity and spontaneous homecage behaviors after a 6hr advance of the dark onset in C3H/HeN mice. LUZ (10mg/kg, s.c.) given for three consecutive days at the new dark onset (8.4±0.4d, n=14) significantly increased the number of days (delay) necessary for re‐entrainment of wheel running activity rhythms compared to vehicle (VEH) controls (6.3±0.3d, n=24; p<0.001). LUZ also delays re‐entrainment of 16 spontaneous homecage behaviors recorded with the HomeCageScan System. The 16 behaviors were grouped into activity, exploratory/anxiety, ingestive and sleep behaviors. LUZ (n=8) compared to VEH (n=8) delayed re‐entrainment to dark onset of the spontaneous behaviors including activity (Walk: VEH, 9.3±o.5d & LUZ, 12.3±0.7d, p<0.005), exploratory/anxiety (Groom: VEH, 9.1±0.6d & LUZ, 12.3±0.6d, p<0.005) and ingestive (Eat: VEH, 9.5±0.5d & LUZ, 12.4±0.6d, p<0.005). We concluded that LUZ acting as inverse agonist at MT1 melatonin receptors may delay the reentrainment of wheel running and homecage behaviors. Support by NS 061068.
In vitro Culture alters the Expression of Xenobiotic Transporters by Mammary Epithelial Cells
Authors: Gehring Ronette , Malreddy Pradeep , Silver Kristopher , Wang Florence , Schultz Bruce
Published: June 2021

Some drugs and toxins are actively transported across the mammary epithelial barrier, resulting in high milk concentrations. To assess potential exposure through milk, in vitro models of the mammary epithelial barrier are needed. The goal of these studies was to compare the expression of xenobiotic transporters between native tissues, cultures of freshly isolated bovine mammary epithelial cells and a bovine mammary epithelial cell line (BME‐UV). Real time PCR was employed to evaluate the expression of mRNA coding for 5 xenobiotic transporters in mammary tissue isolated from lactating dairy cows, primary epithelial cells isolated from these tissues and cultured in T25 flasks or as polarized monolayers on permeable supports, and BME‐UV cells cultured as polarized monolayers on permeable supports. 18S rRNA was employed as an internal standard for comparison. Relative to the native tissue, the expression of mRNA coding for ABCB1 and NCBT1 were increased in all the cell cultures, whereas ABCG2 and OATPD mRNA expression were reduced. The expression of OCT1 mRNA remained unchanged. These initial results suggest that standard culture conditions promote the expression of putative drug transport proteins in proportions and levels that do not directly reflect those observed in the actively lactating dam. Further studies are needed to refine culture conditions for these potential in vitro models. [NIH P20‐RR017686 Core C]
Ovarian cycle‐related effects of neurosteroids on GABA‐A receptor‐mediated phasic and tonic currents in the hippocampus
Authors: Carver Chase , Wu Xin , Murchison David , Griffith William , Reddy Doodipala Samba
Published: June 2021

Endogenous neurosteroids such as allopregnanolone (AP) play a key role in seizure susceptibility because they potentiate GABA‐A receptors in the hippocampus. In this study, we investigated the ovarian‐cycle related changes in δ‐subunit levels and the sensitivity of AP modulation of GABA‐gated phasic currents (IGABA) and tonic currents (Itonic) in the female adult mice hippocampal CA1 pyramidal (CA1) and dentate granule cell (DGC). In immunocytochemistry study, the δ‐subunit staining intensity in DGCs was stronger than in CA1 in diestrus. In isolated whole‐cell recordings, AP (10 nM to 1 μM, n=5 to 10) produced a concentration‐dependent enhancement of IGABA. AP (1 μM) produced a 4.4‐ and 7.1‐fold potentiation of IGABA in CA1 and DGC in diestrus stage, and 3.1‐ and 4.2‐fold potentiation in CA1 and DGC in estrus, respectively. In hippocampus slices, the conductance of Itonic in DGC in the absence and presence of 3 μM GABA were 3.5‐fold and 2.3–fold larger than those in CA1 in diestrus (n=3–12), respectively. Perfusion of AP (300 nM) produced a 0.37‐ and 1.5‐fold potentiation of Itonic in CA1 and DGC of diestrus stage. These currents were completely blocked by the GABA‐A receptor antagonist bicuculline. These studies suggest that cyclic changes of AP‐enhanced IGABA and Itonic in hippocampal neurons are related to δ‐subunit plasticity during the estrus cycle that may affect catamenial seizures. *Supported by NIH NS051398
Prolyl Hydroxylase Domain (PHD)/Hypoxia
Authors: Ebiogwu Adaora , Oyekan Adebayo
Published: June 2021

Hypoxia increases expression of Hypoxia Inducible Factor (HIF) which salvages cells from damage. PHD regulates HIF‐1a activity during hypoxia or normoxia by mechanisms involving nitric oxide (NO) and heme oxygenase (HO). Increased HIF‐1a activity increases sodium (Na) excretion (UNaV) but the ion transport mechanisms involved are not known. Since PHD is expressed predominantly in the medulla, this study tested the hypothesis that PHD is selectively involved in renal sodium transport via Na transporter mechanisms in the medulla. The study evaluated differential sensitivity to the effects of inhibitors of Na transporter mechanisms following PHD inhibition with dimethyloxaloylglycine (DMOG; 15 mg/kg). DMOG increased 24‐hr UV (1.6 fold) and UNaV (3.2 fold). Following acute salt (NaCl) load (2.5 ml/100 g BW), DMOG also increased UV (1.95 fold) and UNaV (1.8 fold). Hydrochlorothiazide (HCTZ, 3 mg/kg), an inhibitor of NCC, Furosemide (FUR, 12 mg/kg), an inhibitor of NKCC), or Amiloride (200 □g/kg), an inhibitor of ENaC, elicited Increases in UV and UNaV that were inhibited by DMOG by 40±4, 39±3, or 34±4%, respectively. DMOG also increased kidney perfusion (Laser scan) by 20±3%, shifted the pressure/natriuresis curve leftwards, and increased UNOxV (2.8 fold) but elicited paradoxical reductions in GFR and plasma bilirubin. These data indicated that PHD participates in renal function to affect tubular function via NO production affecting NCC, NKCC, and ENaC ion transporter mechanisms to a similar extent.