bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of dark chocolate on endothelial function in healthy adults
Authors: Orsa Christina , Plumlee Deva , Wright Alely , Hong Mee Young
Published: June 2021

Cocoa and dark chocolate have among the highest antioxidant concentrations compared to other antioxidant food sources. This study examines if dark chocolate protects against the risk of cardiovascular disease by improving endothelial function and if bloomed chocolate attenuates those effects. Thirty one healthy adults were randomly assigned to consume 50 grams per day of either dark chocolate (70% cocoa), bloomed dark chocolate (70% cocoa), or white chocolate (0% cocoa) for 15 days. A blood lipid profile, glucose, bioelectrical impedance analysis (BIA), blood pressure, and forearm skin blood flow were measured on day 1 and day 15. Dark chocolate consumption did not significantly lower blood pressure, skin blood flow, or body fat. White chocolate consumption for 15 days reduced basal skin blood flow (p=0.010). Both dark chocolate groups, bloomed or not, significantly lowered glucose (p=0.021) and low‐density lipoprotein (LDL) cholesterol (p=0.033) levels and significantly increased high‐density lipoprotein (HDL) cholesterol (p=0.002) levels compared to the white chocolate group. Endothelial function components may improve with a longer duration of dark chocolate consumption. A larger and long term study with dark chocolate consumption remains to be needed. Supported by SDSU UGP and NUTR 302L class.
Nutritional Status for less‐explored micronutrients in underprivileged Guatemalan preschool children: Vitamins D
Authors: Hernández Liza , Solomons Noel W. , Romero-Abal María-Eugenia , de María Gamero Flor , Armas Laura , Bonorden Melissa J.L. , Herreid Richard M.
Published: June 2021

Background

Whereas iron, vitamin A, iodine, zinc and folate are prominent on the public health nutrition agenda, vitamin D has been lower on the interest horizon. Particular dietary or environmental characteristics of Guatemala may predispose to insufficient or deficient nutrient status.

Objective

To assess vit D status in a convenience sample of preschoolers in disadvantaged urban and rural communities receiving subsidized meals.

Methods

104 serum samples were obtained from boys and girls meals in 3 sites in the Central Highlands of Guatemala, 47 from 2 urban (U) centers and 57 from a rural (R) center. Serum 25(OH)D was measured on a Liaison instrument (DiaSorin, Inc., Stillwater, MN) using a chemiluminescent assay in Omaha, Nebraska. Respective cutoff criteria for deficient and insufficient for 25(OH)D were <20 and 20–30 ng/mL. The contribution of dietary vit D from center menus was estimated.

Results

Overall mean 25(OH)D concentration (ng/mL) was 26±6, 27±6 (U) and 25±7 (R) (p>0.05). Globally, 20.2% had deficient values and 56.7% had insufficient values. Deficiency rates by setting were 12.7% (U) and 26.0% (R), and insufficiency rates, 59.6% (U) and 54.4% (R). The 1 or 2 meals offered by the centers provided averages of 0.12–0.90 μg vit D/d.

Conclusion

Sub‐adequate vit D status is a common feature of preschoolers in day‐care center populations, without geographic distinction, related in part to low dietary intakes.
Assessment of actual nutrition of workers in the uranium quarring regions
Authors: Abduldayeva Aigul , Bekenova Farida
Published: June 2021

Rational and balanced nutrition practice is not only an important determinant of health, but also a major factor in preventing and reducing the risk of chronic no communicable diseases. Eating disorders are the leading factors in the development of most non‐communicable diseases, so this problem should be considered with health and social care, hygiene and preventive points of view. In connection with aforesaid it is necessary to study features of workers’ nutrition, to establish the factors and conditions, which influence on them, to exploit corresponding recommendation by optimization. Workers aged 35 – 50 took part in investigation of hygienic estimation of actual nutrition. In result of investigation of nutrition it was determinated that energetically asset of workers’ ration was lower than recommended standard, in average 2558 ±59 kkal. Consumption of proteins composed 86±2 g/day, for the account of them 14 % of the general caloric content of a ration are satisfied. The revealed misbalance in nutrition is essential factor of development risk of functional breaking of workers and demand obligatory correction.
Formative research on hygiene behaviors and geophagy as part of interventions to improve infant growth
Authors: Ngure Francis Muigai , Humphrey Jean H , Mbuya Mduduzi N. N. , Mutasa Kuda , Majo Florence , Chasekwa Bernard , Prendergast Andrew , Curtis Valerie , Stoltzfus Rebecca J.
Published: June 2021

The predominant pathways for fecal‐oral transmission of bacteria for infants may differ from adults and these have not been studied. We conducted direct observation of 23 caregiver‐infant pairs for a total of 130 hours and recorded WASH related behaviors. We identified “potential key vectors” for fecal‐oral transmission of bacteria and enumerated Escherichia coli, Total coliforms, Enterobacteriaceae and total aerobic counts. In addition to infant's fingers, food, and drinking water, three infants actively ingested soil 11.3 ±9.2 times (mean ±SD) and two ingested chicken feces 2± 1.4 times in 6 hours. Infants’ hands were rarely washed and never with soap unless as part of a bath. Drinking water was contaminated with E. coli in 12/22 (54.5 %) households, while no infant food samples tested positive for E coli. Quantitatively, a year old infant ingesting 1 gram chicken feces in a day, 20 grams of soil from a laundry area of the kitchen yard and 400 ml of contaminated water consumes 4,700,000 to 23,000,000, 440 to 4,240 and 400 to 1,200 E. coli counts, respectively, from these sources. These findings suggest that predominant pathways for fecal‐oral transmission of bacteria among infants include direct ingestion of soil and chicken. Behavior change education should include breaking socio‐cultural beliefs that soil ingestion is beneficial for infant health, which are common throughout the world.

Grant Funding Source : Department for International Development (DFID), United Kingdom
Beneficial effects of Korean traditional diets in patients with hypertension and type 2 diabetes
Authors: Jung Su-Jin , Park Soo-Hyun , Choi Eun-Kyung , Kim Min-Gul , Cha Youn-Soo , Song Won O , Park Tae-Sun , Ko Jae-Ki , Chae Soo-Wan
Published: June 2021

The rapid rise in the prevalence of metabolic syndrome,hypertension and diabetes in Korean has paralleled with adoption of western dietary patterns that are rich in animal foods and saturated fat, while replacing Korean traditional diets that emphasize vegetables and fermented foods. We aimed to investigate the efficacy of Korean traditional diets in controlling blood glucose fluctuation, blood pressure, and cardiovascular risk factors in hypertensive and type 2 diabetic patients (n=41, 61.8±1.5yrs) who were taking medications prescribed for respective diseases in a 12‐wk controlled parallel clinical trial. The control group (n=20) was told to “eat as usual” whereas the intervention group (n=21) was fed daily Korean traditional diets for 12 wks. At the end of the trial, both groups had lower BMI, % body fat and WHR compared to those at the baseline (p<0.05). Compared to the control group, the intervention group had a greater % change (p<0.05) from the baseline for HbA1c (−3.4% vs −10.3%), pulse rate (2.3% vs. −8.8%),Valsalva score (0.17 vs. − 0.15) and gamma‐glutamyl transferase (16.4% vs −24.2%). Regular consumption of Korean traditional diets for 12 wks by hypertensive and diabetic patients showed favorable changes in glycemic control, body composition, pulse rate and cardiovascular risk factors.
Relationship between inflammation, oxidative stress, and oxidative damage with severity of knee osteoarthritis (OA)
Authors: Browne Jenna Lee , Hooshmand Shirin , Elam Marcus , Feresin Rafaela , Arjmandi Bahram
Published: June 2021

OA is a disease that is characterized by the progressive degradation and loss of articular cartilage. The etiology of OA is complex; however, it is well known that advanced age is a major risk factor for developing OA. Evidence suggests a role for inflammation, oxidative stress and damage in the progression of OA, all of which are attributable to age‐related changes in cartilage. In OA, a cascade of inflammatory events is initiated by inflammatory cytokines, thereby increasing activity of reactive oxygen species (ROS) and accelerating oxidative damage leading to cell death. Furthermore, free ROS cause damage to DNA. The purpose of this study is to evaluate the extent to which serum and synovial fluid indicators of inflammation (C‐reactive protein, CRP), oxidative stress (malondialdehyde production, MDA) and oxidative damage to DNA (7, 8‐dihydro‐8‐oxo deoxyguanosine, 8‐OHdG) are related to OA severity. Blood and synovial fluid samples were obtained from a total of 40 men and women with knee OA undergoing arthroscopic or total knee replacement surgery. CRP, MDA, and 8‐OHdG are being analyzed using ELISA method. Although results are yet to be analyzed, we hypothesize that increased levels of CRP, MDA, and 8‐OHdG correlate with OA severity.
Dose‐response effect of hazelnut consumption on body composition and inflammatory markers in overweight and obese individuals
Authors: Tey Agnes Siew Ling , Brown Rachel , Gray Andrew , Chisholm Alexandra , Delahunty Conor
Published: June 2021

Recent evidence suggests regular nut consumption may have beneficial effects on cardiovascular disease risk factors that go beyond their cholesterol‐lowering properties, e.g. a reduction in inflammation. Inflammation is more pronounced amongst overweight and obese populations. Current guidelines in New Zealand recommend the daily consumption of 30g of nuts. Studies are lacking regarding the quantity of nuts required to confer anti‐inflammatory properties without compromising the body weight. Thus, the objective of this study was to compare the effects of incorporating hazelnuts at two different doses with a diet without nuts on body composition and inflammatory markers in 110 overweight and obese individuals. The study was conducted using a randomised, controlled, parallel design including three treatment arms: 30 g/d of hazelnuts, 60 g/d of hazelnuts or no nuts (control group) for 12 weeks. Body composition and inflammatory markers were assessed at baseline, 6 weeks and 12 weeks. There was no evidence of any differences in follow‐up measurements between groups after adjusting for baseline values. Hazelnut consumption does not adversely affect body composition but appears to have no effect on inflammatory markers in overweight and obese individuals.
RNA quality analysis of tumor bank specimens and downstream applicability
Authors: Newell Jordan Mark , Patrick Susan , Clawson Gary
Published: June 2021

Quality of biorepository specimens is a topic of concern. The goal of this project was to evaluate the quality and quantity of RNA obtained from biobanked fresh frozen (FF) and matched formalinfixed paraffin‐embedded (FFPE) tissues. Initially, three human cancer cell lines were used, and then ~50 FF biorepository cancer cases (melanoma, colon, pancreatic) were selected. Histologically similar FFPE tissues were then selected from the original surgical cases. RNA was extracted, quantified with a Nanodrop, and RNA Integrity Number (RIN) was determined using an Agilent Bioanalyzer. Sample RINs generally aligned into 2 groups; ~75% of specimens showed relatively high RINs (90% of melanomas showed RINs of ~8, 70% of colorectal cancers showed RINs of ~6, and 80% of pancreatic cancers showed RINs of ~5). ~25% of samples showed poor RINs (~2–3). RINs were uniformly lower (~2–3) for the matched FFPE samples. Age of FF or FFPE specimens was not related to its RIN. RNA (select cases) was then further analyzed using SA Biosciences housekeeping gene PCR Arrays. dCt (Ct FFPE – Ct FF sample; Ct = cycle threshold) was determined for the 12 housekeeping transcripts. Results showed considerable differences in the dCts between housekeeping genes, and suggested that b‐actin (large dCt) may be useful as a “canary” for predicting suitability for downstream expression analyses. Supported by the GCRF and a Dept. Research Initiation grant.
Bryostatin‐1 vs. TPPB: A direct comparison of APP processing and PKC activation in neuronal cells
Authors: Yi Ping , Castor Trevor P , Schrott Lisa , Alexander J Steven
Published: June 2021

The activation of the alpha‐secretase pathway appears to be a critical therapeutic advance in Alzheimerˈs disease. There are two important classes of agents which can activate this pathway, the benzolactam/indolactams and bryostatin/bryologues. Both appear to modulate APP processing through PKC dependent pathways, but a direct comparison of these two classes in a neuronal cell line has not been performed. In this study, SH‐SY5Y neuroblastoma cells were selected as a model and dilutions of Bryostatin‐1 or TPPB were added to serum‐free culture medium to final concentration for 3h, except in time‐course experiments. After treatment, alpha‐secretase activity and translocation of PKC isoforms were detected by western blotting. Our results showed Bryostatin‐1 induces s‐APPa release in SH‐SY5Y cells at several orders of magnitude lower concentration, and at earlier time points than TPPB. We also found Bryostatin‐1 activates PKC‐a, PKC‐d and PKC‐e translocation in SH‐SY5Y cells at 10‐8M, 10‐9M and 10‐10M and different time points, howere TPPB only induced PKC‐a translocation at 10‐9M at 3h. Bryostatin‐1 thus appears to exert a more rapid, potent and sustained activation of APP processing which is associated with its much more potent and specific activation of PKC‐d and PKC‐e at lower doses.

This research was supported by a grant from the NIA and NIH (1R44AG034760‐01A1)
The Wisconsin Respiratory Symptom Survey‐21 to assess health‐related quality of life in college students during upper respiratory infections
Authors: Smith Tracey J. , Rigassio-Radler Diane , Touger-Decker Riva , Denmark Robert
Published: June 2021

The Wisconsin Upper Respiratory Symptom Survey‐21 (WURSS‐21) assesses symptom severity and functional status during upper respiratory infections (URIs), which are important measures of health‐related quality of life (HRQL). To determine the reliability of the WURSS‐21 in college students; and, to explore which URI symptoms and functional impairments contributed to the total severity score. This descriptive study was part of a prospective, 12‐week trial in undergraduate students (n = 198, Framingham, MA). Incidence of URIs, and HRQL during URIs, was determined via the WURSS‐21 (items ranked on a 7‐point scale, 1 = very mild and 7 = severe). Cronbach's α was calculated for survey subscales (nasal symptoms, throat symptoms and functional tasks) using day 3 scores. Sixty‐one percent of subjects experienced ≥1 URI (167 total URIs; 1003 sick days; 993 complete surveys). Cronbach's α was ≥ 0.90, indicating good reliability. The median total WURSS‐ 21 score was 66 (range = 5–832); most items were ranked as mild to moderate. The most frequently reported symptoms were ‘runny nose’ and ‘tired’ (~71% of surveys); the most frequently impaired functional tasks were ‘breathe easily’ and ‘sleep well’ (57% and 49% of surveys, respectively). The WURSS‐21 reliably assessed HRQL during URIs in college students; and, both symptoms and impaired functional tasks contributed to total scores.

Research Funded by Chr Hansen A/S
Role of HMGB1 in Doxorubicin‐induced myocardial apoptosis and its regulation pathway
Authors: Yao Yongwei , Zhang Guohui , Zhang Yingyu , Rui Tao
Published: June 2021

The underlying mechanism by which Doxorubicin (DOX) causes cardiomyocyte apoptosis is not clear. The aim of present study is to investigate the role of high‐mobility group box 1 (HMGB1) in DOX‐induced myocardial apoptosis, and signal pathway involved in regulation of HMGB1 expression in cardiomyocytes with DOX.

Methods and Results

Treatment of cardiomyocytes and naive mice with the DOX resulted in an increased HMGB1 expression which was associated with increased myocardial cell apoptosis. Pharmacological (A‐box) or genetic blockade (TLR4 deficiency, TLR4−/−) of HMGB1 attenuated the DOX‐induced myocardial apoptosis and dysfunction. In addition, DOX resulted in an increment in the generation of peroxynitrite (ONOO‐) and an elevation in phosphorylation of c‐Jun N‐terminal kinase (JNK). Pretreatment of myocytes with FeTPPS, a peroxynitrite decomposition catalyst prevented DOX‐induced JNK phosphorylation, HMGB1 expression, myocardial apoptosis and cardiac dysfunction. Genetic (JNK−/−) or pharmacological (SP600125) inhibition of JNK ameliorated the DOX‐induced HMGB1 expression and diminished myocardial apoptosis and cardiac dysfunction.

Conclusion

the results indicate that HMGB1 mediates the myocardial injury induced by DOX and ONOO‐/JNK is a key regulatory pathway of myocardial HMGB1 expression induced by DOX. (Supported by National Science Foundation of China# 81170210).
The Cell Biology of Neutral Lipid Synthesis and Storage
Authors: Walther Tobias
Published: June 2021

Neutral lipids, such as triacylglycerides, are stored by organisms as precursor for membrane components and as compact reservoirs of metabolic energy. Within cells, neutral lipids are packaged into cytoplasmic lipid droplets (LDs). In these organelles, the core of neutral lipids is bounded by a monolayer of phospholipids into which a set of specific proteins are integrated. Here, we discuss the mechanisms on neutral lipid synthesis and how cells package to resulting triacylglycerides into LDs.
Nicotinic acetylcholine and dopamine receptors contribute to the discriminative stimulus effects of nicotine in nonhuman primates
Authors: Blaylock Brandi Lyn , Nader Michael A
Published: June 2021

Nicotinic acetylcholine receptors (nAChRs) are potential therapeutic targets for neurological disorders for their ability to indirectly modulate neurotransmitter activity including dopamine (DA). The present study investigated the role of nAChR (α4β2, α3β4, α7) and DA D2‐like receptors (D2, D3) in the discriminative stimulus (SD) effects of nicotine (NIC) in cynomolgus monkeys (n=4) trained to discriminate NIC (0.3 mg/kg, i.m., 10‐min pretreat) from saline under a fixed‐ratio 30 schedule of food reinforcement. We found that neither the α4β2 nAChR partial agonist varenicline (0.03–0.3 mg/kg, 1‐h pretreat, i.m.), nor the α3β4/α4β2 nAChR partial agonist cytisine (0.1–1.0 mg/kg, 10‐min pretreat, i.m.) engendered NIC‐like SD effects, but both compounds antagonized the SD effects of NIC. While the peripheral nAChR antagonist hexamethonium (3.2–5.6 mg/kg, i.m.) did not alter the SD effects of NIC, the nonselective nAChR antagonist mecamylamine (0.56–1.7 mg/kg, i.m., 30‐min pretreat) and the competitive α4β2 nAChR antagonist DHβE (0.1–1.0 mg/kg, i.m., 10‐min pretreat) attenuated NIC discrimination. The D2‐like antagonist haloperidol (0.001–0.1 mg/kg, 10‐min pretreat) antagonized NIC. D3 receptors may contribute as the partial agonist PG 619 (0.03 mg/kg, i.m., 10‐min pretreat) attenuated NIC's SD effects. These findings suggest α4β2 nAChRs and DA D2‐like receptors contribute to the SD effects of NIC. DA12460
Methamphetamine‐Induced Locomotor Sensitization in C57BL/6 Mice Requires the MT1 Melatonin Receptor
Authors: Hutchinson Anthony J , Ma Jason , Hudson Randall L , Dubocovich Margarita L
Published: June 2021

Genetic deletion of both MT1 and MT2 melatonin receptors abrogates the development and expression of locomotor sensitization in melatonin proficient C3H/HeN mice repeatedly pretreated with methamphetamine (METH) (Hutchinson et al., Program 669.12. 2010 Neuroscience Meeting Planner). However, C57BL/6 mice deficient in pineal melatonin do not develop or express sensitization using the same METH treatment paradigm. Here we assessed the role of MT1 melatonin receptors in locomotor sensitization using a two METH injection protocol. Mice were pretreated with either vehicle or METH (1.2 mg/kg, i.p.) on Day 1, then challenged on Day 8 with METH (1.2 mg/kg, i.p.). On Day 1 METH increased locomotor activity in both WT [60.6±5.5 meters traveled (n=12), p<0.05] and MT1KO mice [57.1±6.5 m (n=12), p<0.05] when compared to vehicle‐treated controls [15.6±4.4 m (n=12), 11.9±1.5 m (n=12) respectively]. METH challenge triggered a sensitized response in METH‐pretreated WT mice [106.0±5.2 m (n=12), p<0.05], but not in METH‐pretreated MT1KO mice [57.1±6.5 (n=12), p>0.05] compared to corresponding vehicle‐pretreated controls [63.2±7.5 m (n=12), 53.1±5.0 m (n=12) respectively]. Our results indicate that in C57BL/6 mice, the expression of the MT1 receptor is required for the induction of METH‐induced sensitization after a single METH pretreatment. Supported by DA021870 to MLD.
Active immunopharmacotherapy for methamphetamine reduces self‐administration in rats
Authors: Miller Michelle L , Moreno Amira Y , Vaillancourt Brittani D , Wright Jerry , Aarde Shawn M , Creehan Kevin M , Vandewater Sophia A , Janda Kim D , Taffe Michael A
Published: June 2021

At present, there are no approved pharmacotherapies for methamphetamine addiction and existing therapies have limited effect. Recent successes in early clinical trials using immunotherapeutic approaches for cocaine and nicotine addiction have motivated interest in creating similar approaches for methamphetamine (METH) addiction. The current study investigated the effects of a candidate vaccine (MH6) shown to produce METH specific antibodies in mice on METH self‐administration in Sprague Dawley rats. Groups of rats were pretreated with MH6 (n=18) or control vaccine (KLH; n=18) during a 13‐week dosing regimen. This consisted of a priming dose followed by 4 booster doses at 2, 5, 9, and 13 weeks after the prime. Stable, high antibody titers were produced. Subsequently, the effects of vaccination on METH self‐administration were assessed across a range of doses (0.0, 0.01, 0.05, 0.10, 0.15, and 0.25 mg/kg/inf). In general, vaccinated rats self‐administered less METH than non‐vaccinated rats, although different patterns were observed depending on whether the rats were pellet trained prior to initiation of the self‐administration procedure. These data provide evidence that active immunopharmacotherapy can provide functional protection against the behavioral effects of METH.
Acute noxious stimulus differentially alters morphine‐induced operant behavior in male and female mice
Authors: Neelakantan Harshini , Walker Ellen A
Published: June 2021

Fewer studies measure pain‐depressed relative to stimulus‐evoked behaviors in preclinical rodent pain models and even fewer studies use both male and female mice. The purpose of the present study is to test the hypothesis that the presence of an acute noxious stimulus will decrease food‐maintained operant behavior and reduce the potency of morphine to produce discriminative stimulus effects. Firstly, mice were trained to respond on a fixed ratio schedule to 50% ensure solution. In this assay, acetic acid (0.2–0.4%) produced concentration‐dependent suppression of responding in mice that was dose‐dependently prevented by morphine (0.32–3.2 mg/kg). Subsequently, male and female C67Bl/6 mice were trained in a two choice operant procedure to discriminate 3.2 mg/kg morphine or saline following which the dose response curves were generated in the absence and presence of 0.4% acetic acid. Morphine produced dose‐dependent drug‐appropriate responding in both male and female mice with no observed potency differences. However, male mice were significantly less sensitive to morphine and acetic acid‐induced effects on operant response rates. Importantly, the presence of acute noxious stimulus failed to alter the discriminative effects of morphine in female mice but significantly attenuated in male mice. In summary, pain‐depressed operant behavior and morphine‐induced discriminative effects may be differentially modulated in male and female mice suggesting the importance of sex differences in the clinical effectiveness of opioids and the abuse liability of morphine in the presence of pain. (Supported by CA129092)
Use of RFID Tags to Capture Dynamic Behavior of Individual and Groups of Small Fish
Authors: Pritchard Josh , Zurn Jane Brooks
Published: June 2021

We describe a system by which we can implant RFID tags into small fish (such as carassius auratus) to use with readers to capture group dynamics as they interact with a variety of standard experiments including the signaled avoidance paradigm with an aquatic shuttle box, and operant responding on a variety of schedules. This innovative approach will allow for fish to be housed in groups, as well as used to determine what individuals do within group behavior. Instrumentation provided by Florida Research Instruments, Inc.
Interactions between the orphan receptor GPR37L1 and the dopamine D1 receptor
Authors: Meyer Rebecca , Hall Randy A.
Published: June 2021

Studies on receptor‐receptor interactions between GPCRs provide valuable insight into crosstalk among different signaling pathways and also indicate potential novel pharmacological targets. The dopaminergic system is of particular pharmacological interest due to its importance in numerous disorders in the central nervous system, including drug abuse and Parkinson's disease. Our lab has been investigating the modulation of dopaminergic receptors through interactions with the orphan GPCR sub‐family GPR37 and GPR37L1. These receptors are highly expressed in dopaminergic areas of the brain and previous work from our lab has indicated that GPR37 is able to physically interact with dopaminergic D2 receptors to alter D2 receptor pharmacological properties. In new studies, we have found that GPR37 does not associate with dopamine D1 receptors; however the closely related GPR37L1 shows a robust D1 receptor interaction. This results in significant changes in D1 receptor function, including alterations in G protein coupling and other signaling properties. In ongoing studies, we are further exploring this novel receptor‐receptor association and its role in both normal and aberrant dopaminergic signaling. This work was supported by NIH R01 NSO55179.
Anxiolytic action of pterostilbene: involvement of hippocampal ERK phosphorylation
Authors: El-Alfy Abir T , AlRahim Md , Silistreli Kalpten , Rimando Agnes M
Published: June 2021

The primary objective of this study was to evaluate the anxiolytic effect of the natural product pterostilbene. Adult male mice administered pterostilbene (1–10 mg/kg, p.o.) were subjected to the elevated plus maze (EPM) test. Pterostilbene manifested anxiolytic activity at 1 and 2 mg/kg doses, demonstrated by increases in % permanence time and number of open arm entries. In addition, the anxiolytic activity of pterostilbene was comparable to that of diazepam in the EPM. The locomotor activity of the animals was unaffected at all doses. Western blot analysis revealed a decrease in both ERK1 and ERK2 phosphorylation in hippocampal homogenates from mice treated with 1 and 2 mg/kg ptserostilbene. Moreover, pterostilbene was detected in the sera and brains of mice following single oral administration. Neither anxiolytic nor motor activity impairment was observed at the higher doses (5 and 10 mg/kg). These results suggest that pterostilbene has the potential to become a candidate compound for therapeutic drug development for anxiety disorders.
Using cocaine self‐administration to determine whether dopamine receptor antagonists are potential substrates for p‐glycoprotein‐ like efflux transporters in the rat blood‐brain barrier
Authors: Norman Andrew B. , Tabet Michael R. , Norman Mantana K. , Tsibulsky Vladimir L.
Published: June 2021

Among a series of dopamine receptor antagonists the in vivo potency of spiperone is less than (−)eticlopride despite having higher in vitro affinity. It was hypothesized that spiperone, but not eticlopride, was a substrate for tamoxifen‐sensitive drug efflux transporters that form part of the blood‐brain barrier (BBB). In rats that were trained to reliably self‐administer cocaine the cumulative level of cocaine at the time of each lever press (satiety threshold) was calculated during the maintenance phase of each session. Rats were then injected i.v. with an antagonist along with tamoxifen or its vehicle and the session continued. Each antagonist maximally increased the satiety threshold approximately 3‐fold, but tamoxifen itself had no effect. In the presence of tamoxifen (5,000 nmol/kg) the effect of eticlopride (20 nmol/kg) was not altered and there was a modest 15% increase in the magnitude of effect of haloperidol. In contrast, in the presence of tamoxifen there was an 80% greater spiperone (75 nmol/kg)‐ induced increase in the maximal cocaine satiety threshold. These data suggest that the low in vivo potency of spiperone is due, at least in part, to its being a substrate for p‐gp‐like efflux transporters in the BBB, which normally reduce brain concentrations of this antagonist. This method provides a useful in vivo assay of whether drugs are substrates for efflux transporters. Supported by NIH grant DP1DA031386.
Transporter‐mediated mechanism of nucleoside penetration of the blood‐testis barrier
Authors: Klein David Michael , Evans Kristen K. , Hardwick Rhiannon N , Dantzler William H , Wright Stephen H , Cherrington Nathan J
Published: June 2021

The blood‐testis barrier (BTB) prevents the entry of many therapeutics into the lumen of the seminiferous tubules (STs) shielding semen from chemical exposure. The BTB is formed by tight junctions between Sertoli cells. One drug class of HIV therapeutics, nucleoside analogs (NSAs), can penetrate the BTB and be detected in the semen. The purpose of this study is to determine the mechanisms by which NSAs are transported across the BTB. Transport studies in isolated rodent STs using H3uridine (Kt 89.72uM) as a model nucleoside substrate indicate that Sertoli cells take up nucleosides almost exclusively via equilibrative nucleoside transporter 1 (ENT1). The IC50 for NBMPR, an ENT inhibitor, is 12.9 nM. These data correspond with immunohistochemical staining of rat testes showing ENT1 on the basolateral membrane, whereas ENT2 is on the apical membrane of Sertoli cells. This localization suggests that ENT1 acts as an uptake transporter and ENT2 may facilitate the efflux of NSAs into the lumen of STs. Uptake of didanosine, a commonly used NSA, in HEK293 cells can be blocked by ENT1 inhibition. Trans‐epithelial transport of uridine by primary rat Sertoli cells can also be blocked by NBMPR. These data indicate a novel ENT1 and 2‐dominant mechanism for the transepithelial transport of nucleoside analogs across the BTB. AI083927.
NO donors subunit and state‐dependently increase desensitization of acid sensitive ion channels
Authors: Bolshakov Konstantin , Dorofeeva Natalia , Swain Sandip , Nikolaev Maxim , Potapjeva Natalia , Bera Amal Kanti
Published: June 2021

Nitric oxide donors were studied on acid‐sensing channels (ASICs) of rat striatal neurons and CHO cells expressing recombinant receptors. S‐nitroso‐acetyl‐penicillamine (SNAP) potentiate or inhibit ASICs depending on their functional state. It potentiated ASICs after pretreatment with extracellular pH 7.4. When ASICs were partially desensitized by pretreatment with pH 7.0, application of SNAP led to increase of their desensitisization. The potentiating effect of NO on ASICs is explained by an increase of ASIC proton sensitivity, whereas inhibition of ASICs by NO is due to an increase of their steady‐state desensitization. Potentiation and inhibition produced by SNAP had different subunit dependence. The most sensitive to potentiation were ASIC1a/2a and ASIC2a channels while ASIC2a and ASIC3 were the most sensitive to inhibition. Potentiation, produced by SNAP, was prevented by sulphhydryl reagent mercaptoethanol, but it did not affect inhibition produced by SNAP. Thus potentiation and inhibition seems to be mediated by different binding sites. We suppose both effects of SNAP are most probably due to nitrosylation, but not to soluble guanylate cyclase activation, because they were unaffected by ODQ, inhibitor of NO‐sensitive guanylate cyclase, and by 8‐Br‐cGMP, an activator of cGMP‐dependent PKG cascades. Work was supported by grant RFBR/STGI 09‐04‐92662‐Ind‐a.
Differential effects of PI3K isoforms on TRPC channels in vascular smooth muscle: Novel actions of PI(3)P‐containing molecules
Authors: Albert Anthony Paul , Shi Jian , Ju Min , Large William
Published: June 2021

It is thought that G‐protein‐coupled receptors linked to phospholipase‐mediated pathways and generation of diacylglycerol (DAG) activate TRPC1 and TRPC3/C6/C7 channel subtypes in vascular smooth muscle cells (VSMCs), via respectively PKC‐dependent and –independent mechanisms. However, endothelinA receptor (ETA) stimulation of TRPC1/C5/C6 and TRPC3/C7 channel activities in rabbit coronary artery VSMCs are not blocked by phospholipase inhibitors. The aim of this work was to study activation mechanisms of these ETA receptor‐induced TRPC channels using patch clamp techniques. ETA receptor‐evoked TRPC1/C5/C6 and TRPC3/C7 channel activities were both inhibited by low concentrations of wortmannin. In addition, TRPC1/C5/C6 channel activity was inhibited by the pan‐Class I PI3K isoform inhibitor ZSTK474, and the Class I PI3Kgamma isoform‐selective inhibitor AS252424. Interestingly, ZSTK474 and AS252424 both increased ETA receptor‐evoked TRPC3/C7 channel activity. Substrates and products of PI3K‐mediated pathways activated TRPC1/C5/C6 channels. In contrast, only PI(3)P activated TRPC3/C7 channels, whereas substrates and other products of PI3K‐mediated processes inhibited these channels. These results show for the first time that PI3K‐mediated signalling pathways have a pivotal role in regulating native TRPC channels in VSMCs. This work was supported by the British Heart Foundation.
Sodium channel activator‐stimulated neuronal development involves BDNF‐TrkB signaling
Authors: Jabba Sairam V , Murray Thomas F
Published: June 2021

N‐methyl D‐aspartate receptor (NMDAR) activation stimulates calcium influx and brain‐derived neurotrophic factor (BDNF) – TrkB signaling. This pathway is implicated in activity‐dependent neuronal development and synaptic plasticity. Voltage‐gated sodium channel (VGSC) activators promote neuronal development by increasing [Na+]i and upregulating NMDAR function. Here we tested the effect of the VGSC activator veratridine (VRT) on neurite outgrowth (NOG), synthesis and release of BDNF and TrkB activation in DIV1 cerebrocortical neurons. VRT enhanced NOG in a hormetic concentration‐response manner and this response was dependent on NMDAR and TrkB signaling. Inhibitors of NMDAR, TrkB, PI3K, and PLCγ inhibited VRT‐enhanced NOG. Acute treatment with VRT stimulated phosphorylation of TrkB and its downstream effectors Akt, mTOR, PLC, ERK1/2 and CREB. VRT increased BDNF synthesis and release in a concentration dependent manner; however, VRT stimulation of TrkB phosphorylation displayed a biphasic concentration‐response curve. VRT stimulation of BDNF synthesis required VGSCs and NMDARs. These data suggest that BDNF synthesis and release mediate VGSC activator‐enhanced NOG through activation of TrkB signaling.

(NIH, R01NS053398)
Mrp1 protects against Doxorubicin induced cardiotoxicity
Authors: Coy Donna J , Jungsuwadee Paiboon , Noel Teresa , Chaiswing Luksana , Oberley Terry , St. Clair Daret , Vore Mary
Published: June 2021

Multidrug resistance‐associated protein 1 (Mrp1) is expressed in the sarcolemma of murine heart and effluxes endo‐ and xenobiotics. Doxorubicin (DOX) is a chemotherapeutic drug known to cause cardiactoxicity in cancer patients through oxidative stress. DOX significantly increases Mrp1 in whole heart, submitochondrial fractions, and mitochondria. Mrp1 is adversely affected during oxidative stress by adduction with 4‐hydroxy‐2‐nonenal. We postulate that Mrp1 serves a protective role in the heart by effluxing toxic products of oxidative stress. Wild type C57BL/6J (WT) or Mrp1 knock out (KO) mice were treated with DOX or saline and sacrificed at 24 or 72 hours. Heart tissue slices were used for electron microscopy and glutathione (GSH) and glutathione disulfide (GSSG) measured by HPLC. At 24 and 72h, significantly increased pathologic changes (p<0.05) were seen in cytoplasm and mitochondria of WT and KO mice, with significantly greater damage in mitochondria in both groups. At 72h, Mrp1 KO mice showed evidence of cytoplasmic edema and apoptotic nuclei (13%). GSSG levels were increased in KO mice at 72h (p<0.001). No changes were observed in GSH levels. The GSH to GSSG ratio was decreased in KO mice at 72h, indicating oxidative stress. These results indicate increased DOX induced cardiotoxicity in Mrp1 KO mice. Further studies are needed to elucidate the protective role of Mrp1 in cardiac tissue. NCI(1RO1CA139844‐D2)
4‐Hydroxy‐2‐nonenal‐ Modified Proteins in Bone Marrow of Phenol/Hydroquinone Treated Rats: Implications for Benzene‐Mediated Hematotoxicity
Authors: Kuhlman Christopher Lee , Petersen Dennis R. , Tsaprailis George , Monks Terrence J. , Lau Serrine S.
Published: June 2021

Benzene exposure causes myelotoxicity and leukemia. Phenol and hydroquinone (PHE/HQ; 1.1/0.9 mmol/kg, ip, 19h), metabolites of benzene, significantly decrease lymphocyte counts (58–89%) in rats. HQ readily oxidizes to 1,4‐BQ and conjugates with GSH, and multi‐substituted HQ‐GSHs were detected in bone marrow after PHE/HQ. HQ‐GSHs are more efficient generators of superoxide anion than HQ/1,4‐BQ, and thus retain the ability to arylate proteins and to redox cycle. However, the precise molecular mechanism(s) by which benzene/HQ‐GSH conjugates induce hematotoxicity is unknown. Using anti‐4‐hydroxy‐2‐nonenal (4HNE) specific antibodies on 2D western, and MS/MS peptide sequencing analysis of immunopositive proteins isolated from bone marrow lysate of PHE/HQ treated rats, we revealed the presence of 9 4HNE‐adducted proteins. In particular, protein disulfide isomerase (PDI) and peroxiredoxin 2 (PRDX2) were immunopositive for 4HNE adduction. In models of alcoholic liver disease, both PDI and PRDX6 have reduced activity when adducted by 4HNE. Moreover, a 4HNE adduction site was identified on calreticulin, on a critical cysteine residue involved in its binding to misfolded proteins. Analysis of the structural and functional significance of these protein modifications may provide new insight into the progression of benzene‐induced leukemia. (P30ES006694, RO1GM70890, Sci. Found. of AZ, T32ES007091)
Mechanisms of action of cytotoxicity of transition metal oxide nanoparticles in human lung cells
Authors: Huang Yue-Wern , Chusuei Charles C. , Aronstam Robert S.
Published: June 2021

The cytotoxicity of nano‐sized oxides of fourth period metals (Ti, Cr, Mn, Fe, Ni, Cu, and Zn), which are widely used in industry, was assessed in two human lung cell lines (A549 & BEAS‐2B). The particles were 16–82 nm in diameter. Toxicity expressed on the bases of particle mass increased with atomic number. Metal oxides altered the cellular redox state, leading to cellular injury as revealed by lipid peroxidation, DNA strand breaks, and membrane depolarization. Oxidative stress produced by exposure to metal oxide nanoparticles also perturbed certain cellular signaling pathways, induced the expression of cell death‐related genes, and compromised the ability of cells to maintain a low concentration of intracellular calcium. Synergistic interactions between intracellular [Ca2+] and OS are a likely contributing factor. While [Ca2+] and OS affect the activity of each other, they induce cell death by distinct pathways. We evaluated the contributions of physicochemical properties of metal oxides to the initiation of cellular oxidative stress. Particle surface charge, particle surface available binding sites, and metal dissolution highly correlated with oxidative response. Our results can inform design of safer nanomaterials for applications in biomedical science.
Lipopolysaccharide impairs gap junction function and reduces rat kidney NRK‐52E cell proliferation
Authors: Wang Yanling , Wei Jing , Xia Zhengyuan , Hei Ziqing , Luo Chenfang
Published: June 2021

Endotoxin lipopolysaccharide (LPS)‐induced shock is life‐threatening which is often associated with acute renal failure. Gap junctions (GJ) play critical roles in maintaining kidney functions by facilitating intercellular communication and tubular purinergic signaling. However, it is unknown whether or not LPS may cause acute kidney injury by impairing renal GJ functions. Renal tubular epithelial cells are one of the main parenchymal cells that are susceptible to injurious stimuli. We, therefore, employed normal rat renal tubular epithelial cell line (NRK‐52E) to investigate the impact of LPS on cell proliferation and GJ function. NRK‐52E cells were seeded at high and low density to promote or impede GJ formation, respectively, and to establish distinctive levels of intercellular communication We found that LPS dose‐dependently reduced cell proliferation rate and colony‐formation rate of high and low density NRK‐52E cells when LPS concentration was above 100 ng/mL, accompanied with reduced connexin43 protein expression and reduced fluorescent dye transmission between adjacent cells (all p<0.05 vs. control without LPS stimulation). The GJ inhibitor oleamide attenuated LPS‐induced reduction in colony‐formation rate of high density cells, but further decreased fluorescent transmission (all P<0.05 vs. LPS). In contrast, GJ agonist retinoic acid increased fluorescent dye transmission. It is concluded that impairment in GJ function may represent a mechanism whereby LPS induces kidney cell injury.

Supported by NSFC grant (No. 81170449) and Guangdong Natural Science Foundation grant (S2011020002780).
Characterization of a Cyp2a(4/5)bgs‐null Mouse Model: Role of CYP2A and CYP2B in Nicotine Metabolism
Authors: Li Lei , Wei Yuan , Zhou Xin , Hough Lindsay B. , Ding Xinxin
Published: June 2021

Nicotine metabolism is believed to affect not only nicotine's pharmacological effects, but also nicotine addiction. As a key step toward testing this hypothesis, we have produced knockout mouse models targeting P450 genes possibly involved in nicotine metabolism, including Cyp2a and Cyp2b. Our recent study of a Cyp2a5‐null mouse confirmed that, although CYP2A5 plays a major role in systemic clearance of nicotine and cotinine, other P450 enzymes may also have significant roles. The aim of this study is to determine, using a newly generated Cyp2a(4/5)bgs‐null mouse model, whether hepatic P450 enzymes (other than CYP2A5) encoded by the Cyp2a(4/5)bgs gene cluster, including CYP2A4 and the various CYP2B isoforms, contribute to nicotine metabolism in vivo. We found that, compared to the Cyp2a5‐null mice, the Cyp2a(4/5)bgs‐null mice showed ~60% decreases in hepatic microsomal nicotine oxidase activity in vitro, and in rates of systemic nicotine clearance in vivo. The decreases in nicotine metabolism in the Cyp2a(4/5)bgs‐null mice were accompanied by prolonged nicotine‐induced hypothermia and analgesia, compared to the behavioral responses in wild‐type mice. Thus, both CYP2A and CYP2B contribute to nicotine clearance in mice. The Cyp2a(4/5)bgs‐null and Cyp2a5‐null mouse models will be valuable for studies on the impact of genetic variations in nicotine metabolism, as occur in humans, on nicotine dependence.
AKAP150‐dependent changes in Kv channel expression in ventricular myocytes following myocardial infarction
Authors: Nieves Madeline , Hirenallur-S Dinesh K , Hinke Simon A , Scott John D , Santana Luis F
Published: June 2021

AKAP150 is anchoring protein that plays important role in compartmentalizing protein kinase A and calcineurin signaling to specific sarcolemmal domains in cardiomyocyte. Increased β‐ adrenergic signaling after myocardial infarction (MI) triggers activation of the Ca2+‐dependent phosphatase calcineurin and the transcription factor NFATc3, resulting in down‐regulation of voltage‐gated K+ (KV) channel in ventricular myocytes. Here we tested the hypothesis that AKAP150 targeting of calcineurin to the sarcolemma is required for NFATc3 activation and the subsequent down‐regulation of KV currents observed after MI. We found that ablation of AKAP150 decreased NFATc3 nuclear translocation and was protective against down‐regulation of KV currents after MI. To investigate the role of AKAP150‐associated calcineurin in the regulation of Kv channel expression; we investigated the effects of MI on ventricular myocytes of a mouse genetically engineered to express an AKAP150 that lacks its binding site for calcineurin (AKAP150 Δ CaN). The amplitude of Kv currents was decreased in WT, but not in AKAP150 Δ CaN cardiomyocytes after MI. These data suggest that targeting of calcineurin to the sarcolemma of ventricular myocytes by AKAP150 is critical for the activation of calcineurin/NFATc3 signaling pathway and the down regulation of KV currents after MI.
Cardiovascular function time course in a rodent model of burn injury
Authors: Henderson Kyle K , Barefield David , Sadayappan Sakthivel , He Li-Ke , Szilagy Andrea , Shankar Ravi , Gamelli Richard L , Kennedy Richard
Published: June 2021

Severe burn injury may attenuate cardiovascular function with the diminished function contributing to morbidity and mortality. Male rats were randomly divided into sham (N = 8) or burn (N = 12) groups. Dorsal burns (30% surface area) were induced by immersion in 100°C water for 25 sec. Fluid resuscitation with Lactated Ringers (4ml/kg bodyweight/% burn, i.p.) was administered over 24hrs. Echocardiography assessed left ventricular (LV) structure and function 2, 4, and 7 days after burn. At day 7, LV function was measured with pressure volume catheterization, and serum collected to measure cardiac biomarkers. Echocardiography revealed a progressive increase in cardiac output (CO) secondary to an increase in stroke volume (SV) and LV dilation in burn animals. At day 7, burn animals had greater end diastolic volume (P = 0.01) and CO (P = 0.04), with a trend toward greater SV (P=0.06). Burn animals also had greater LV contractility (+dP/dt: 11319±1013 vs. 8336±411) and rate of relaxation (Tau: 7.9±0.3 vs. 9.4±0.6 msec). Cardiac myosin binding protein‐C was slightly elevated in burn animals (8.30±1.93 vs. 3.23±0.68ng/ml, P = 0.10). These preliminary studies in a rat model demonstrate that a full thickness burn leads to a sustained increase in cardiac function, which parallels the hyperdynamic state previously reported by others. If continued this effect may lead to cardiomyopathy. Funding: USAMRAA W81XWH‐10‐2‐0172.
The role of adrenergic receptor beta 1 in cardiac adrenergic responsiveness during thyrotoxicosis
Authors: da Costa Bocco Barbara Miranda Leite , Lancelloti Carmem Lucia , Christoffolete Marcelo Augusto , Brum Patricia Chakur , Ribeiro Miriam Oliveira
Published: June 2021

Alterations in cardiac functions are observed in patients with thyrotoxicosis, similar to those observed in state of hyper‐stimulation of the Sympathetic Nervous System, although circulating levels of catecholamine are normal, suggesting that the T3 increases adrenergic responsiveness in the heart. To evaluate this hypothesis, we studied the effects of thyrotoxicosis in mice with knockout for beta1 adrenergic receptor (B1KO), the main isoform in the heart. For that, we treated the animals with T3 (60 ng/g BW) for 3 weeks. Our data indicates that the increase in the heart rate induced by T3 in B1KO mice was significantly higher when compared to wild type (WT) and (39.7±2.5 WT+T3 vs. 102.4±8.3 bpm B1KO+T3, p< 0.0001). T3 induced significant hypertrophy in WT mice when compared to control (4.3±1.7 WT vs. 10.5±1.7μm WT+T3, p<0.001) but not in B1KO (8.0±2.3 B1KO vs. 8.0±1.9 μm B1KO+T3). The increase induced by T3 in mRNA levels of SERCA II, α‐MHC observed in WT was abolished in B1KO. The decrease in mRNA levels of β‐MHC induced by T3 was also abolished in B1KO mice. In conclusion, the cardiovascular alterations induced by T3 in thyrotoxicosis are dependent on the Beta 1 adrenergic signaling.