bluuebunny/crossref_metadata_2025 huggingface dataset:
The role of dietary fiber in regulating lipid bioaccessibility of almonds during mastication
Authors: Ellis Peter R , Grundy Myriam , Grassby Terri , Mandalari Giusy , Faulks Richard M. , Berry Sarah E.E. , Waldron Keith W. , Butterworth Peter J.
Published: June 2021

Almonds contain phytochemicals and nutrients that potentially have positive health benefits in relation to heart disease, diabetes and obesity. One important mechanism associated with these benefits is the bioaccessibility (release) of lipids from the almond tissue during mastication and digestion. Indeed, it has been reported that in human subjects on almond‐rich diets, a significant proportion of lipid remains undigested, mainly caused by the entrapment of lipid by intact cell walls (dietary fiber). However, the bioaccessibility of almond lipid has not yet been quantified. We have studied this by performing a mastication trial in healthy human volunteers (n = 15). The particle size distribution of chewed almonds was measured using mechanical sieving and laser diffraction methods. Lipid release from ruptured almond cells was estimated using a theoretical model based on the particle size of the chewed almond tissue and the diameter of the lipid‐rich cells. Our results showed that laser diffraction was the most reliable and efficient method. Lipid release from masticated almonds was estimated to be ~30% of total lipid and originated from the ruptured cells of the fractured tissue. This indicates that the majority of almond lipid is unavailable for the early stages of digestion. Further work will extend to studies of lipid release and digestion at more distal sites of the gastrointestinal tract. This work was funded by the BBSRC, UK (DRINC; BB/H004866/1).
α‐Tocopherol absorption into a three‐dimensional human skin tissue model
Authors: Evans Julie A , Garlick Jonathan , Johnson Elizabeth J. , Wang Xiang-Dong , Chen C-Y. Oliver
Published: June 2021

Background

α‐Tocopherol (AT) is an important antioxidant for skin health. An in vivo‐like model is needed for evaluation of the bioactions of AT.

Objective

This study aimed to determine the feasibility of AT uptake in a 3‐dimensional (3D) human skin model.

Methods and Materials

AT was applied to media (0–25 μM) or topically (0–1 mg/cm2) for up to 48 h. Skin absorption of AT was quantified in dermis and epidermis by RP‐HPLC with UV detection after proteolysis and standard lipid extraction.

Results

Over a 48 h medium incubation with AT (25 μM), dermal and epidermal concentrations of AT increased to approximately 3 and 30 ng/mg wet weight, respectively. Over a 48 h topical incubation with AT (1 mg/cm2), concentrations of AT in the dermis were 0.4 μg/mg wet weight and increased in the epidermis to approximately 27 μg/mg wet weight. After 48 h incubation with 15 μM AT, dermal and epidermal concentration of AT were 1 and 21 ng/mg wet weight, respectively.

Conclusions

Our 3D skin tissue model is a feasible tool to assess AT absorption into dermal and epidermal compartments. AT absorption into this model was time and dose‐dependent with concentrations reaching those found in vivo. This model allows for further investigation into the photoprotection and mechanisms by which nutrition may play a role in skin health. Support: USDA#581950‐7‐07; Almond Board of California
Structure and functions of the fission yeast Swi5‐Sfr1 complex, an activator of Rad51 presynaptic filaments
Authors: iwasaki Hiroshi
Published: June 2021

The fission yeast Swi5‐Sfr1 complex stimulates the intrinsic activity of the Rad51 recombinase to promote DNA strand exchange reactions. Domain analyses revealed that Swi5 in a complex with the C‐terminal domain of Sfr1 (Swi5‐Sfr1C) has an essential activator activity for Rad51‐dependent strand exchange. Interestingly, Swi5‐Sfr1C binds very transiently to Rad51, but the N‐terminal domain of Sfr1 (Sfr1N) binds to Rad51 with much higher affinity. The structures of Swi5, Swi5‐Sfr1 and Swi5‐SfrC have been determined. Swi5 and Sfr1C form a tight 1:1 complex of a parallel coiled‐coil heterodimer that is joined firmly together via two newly identified leucine‐zipper motifs and a bundle. The resultant coiled‐coil is sharply kinked, generating an elongated crescent‐shaped structure suitable for binding within the helical groove of the Rad51 filament. The N‐terminal region of Sfr1 is intrinsically disordered. We suggest that the snug fit resulting from the complementary geometry of Swi5‐Sfr1C activates the Rad51 filament by retaining it as an ATP‐bound form and that Sfr1N plays an anchor role in a transient set‐in of the Swi5‐Sfr1 complex into the filament groove. This transient interaction may be critically important for efficient DNA strand exchange as the groove directly participates in incorporating homologous duplex DNA.
Increased carcinogen‐induced colon cancer in ERβKO compared to Wild Type mice
Authors: MacDonald Ruth S , Przybyszewski Joseph , Lubahn Dennis B
Published: June 2021

Colon cancer risk is influenced by estrogen exposure, although the mechanism through which this occurs is not defined. Conversion of estrogen to estrone appears to be protective in colon and is lost in colon tumors. We hypothesized that dietary phytoestrogens or estrone would reduce colon tumorigenesis via an estrogen receptor beta (ERβ) dependent mechanism. Ovariectomized ERβKO or wild type female mice were fed diets containing casein, soy protein lacking isoflavones (Soy‐IF), soy protein+genistein (Soy+Gen), soy protein+NovaSoy (Soy+NS) or soy protein+estrone (Soy+E1) from weaning. Colon tumors were induced with 6 weekly injections of azoxymethane (AOM). At termination (32–38 weeks after AOM), colon tumor incidence was lower in WT (32%) compared to ERβKO (48%) mice. ERβKO and WT mice fed Soy+Gen had lower tumor incidence than mice fed all other diets. Despite no overall differences in total body weight, mice fed estrone had higher body fat weight than the other groups. No differences in colonic TNFα, IL‐10 and other cytokines due to diet or genotype were found, except IL‐1β was significantly higher in WT mice fed soy‐IF or Soy+NS compared to other diets. This study demonstrates a protective role for ERβ and suggests a protective effect of soy phytoestrogens in colon cancer. The mechanism does not appear to be due to modulation of the colonic immune system.
Quercetin decreases tumorigenesis in a high fat diet‐enhanced mouse model of breast cancer
Authors: Steiner Jennifer , Davis J Mark , McClellan Jamie , Green Jeffrey , Murphy E Angela
Published: June 2021

Consumption of a high fat diet (HFD) increases the risk for breast cancer (BrCa). Bioactive food components such as quercetin (Quer) may target processes relevant to HFD‐enhanced BrCa.

OBJECTIVE

To determine the efficacy of Quer in prevention of HFD‐enhanced tumorigenesis in the C3(1)SV40Tag mouse model of BrCa.

METHODS

Female C3(1)SV40Tag mice were assigned to a dietary treatment group (n=14–15) at 4wks of age: control (CON) diet (AIN76A; 11% kcal fat), HFD (41% kcal fat), or HFD supplemented with 0.02% Quer (QHFD). Body weight (BW) and food intake were measured weekly and palpable tumors bi‐weekly. At 18wks of age mice were sacrificed and all visible tumors were counted, and measured. Tumors were analyzed for mRNA expression of pro‐tumoral factors (VEGF, IL‐6, CD206 and TNF‐á) using RT‐PCR.

RESULTS

HFD intake (HFD and QHFD) significantly increased BW (wks 8–18) (P<0.05), despite no differences in kcal intake. Tumor number and volume were increased following HFD at 18wks and Quer offset this effect (P<0.05). In fact, QHFD mice had significantly lower tumor volume than CON at 18 wks. Tumor expression of VEGF, IL‐6, CD206 and TNF‐á all showed strong trends of an elevation in HFD compared to CON and QHFD.

CONCLUSIONS

These data provide strong support for a beneficial effect of Quer in reducing HFD‐enhanced BrCa that may be mediated by its ability to decrease pro‐tumoral factors within the tumor.

Grant Funding Source : None
Bioactive tomato components inhibit cancer promoting activity of testosterone in the mouse prostate epithelium
Authors: Tan Hsueh-Li , Elsen Amy C. , Erdman John W. , Thomas-Ahner Jennifer M. , Clinton Steven K.
Published: June 2021

We hypothesize that tomato phytochemicals, such as lycopene (LYC), may antagonize cancer promoting testosterone activity in the normal prostate. C57/BL6 male mice (4 wks; n=84) consumed a control AIN93G or an identical diet with 10% tomato powder (TP) or 0.04% LYC beadlets. Mice were randomized to intact, castration, or castration + testosterone at 8 wks. Serum LYC analysis was by HPLC and prostate proliferation (Ki67) and apoptosis (ApopTag) were evaluated. Prostate gene expression was by qRT‐PCR. Serum LYC concentrations were 366 nmol/L for TP and 371 nmol/L for LYC and were reduced by castration (−38% in TP, P=0.02; −42% in LYC, P<0.001). Epithelial proliferation in TP and LYC fed intact mice was reduced by 50% (P<0.001) with a 30% inhibition (TP, P=0.02; LYC, P<0.001) observed in the castrate‐testosterone mice. Castration dramatically enhanced apoptosis which was further increased by TP (2.2%, P<0.05). CMO1 expression is present in the liver and not detected in the prostate. The CMO2 gene, present in the liver and potentially involved in eccentric LYC cleavage, was detected in the prostate and reduced by castration (P=0.005) with no effect of diet. Tomato phytochemicals may inhibit early prostate carcinogenesis by impacting androgen signaling pathways that impact sensitivity to early steps in the carcinogenic process. Testosterone may impact CMO2 gene expression suggesting a diet and endocrine interaction.

Grant Funding Source: NIH‐NCI/AICR
Polyphenolic and Antioxidant Properties of Elderberry (Sambucus) Fruits
Authors: Finn Amy Marie , Milliron Sidney E , Hummer Kim E , Wilson Ted
Published: June 2021

Elderberries (Sambucus sp.) are rich in phenolics which could provide protection of LDL against oxidation. S. Canadensis (S.c.) and S. nigra (S.n.) are native to N American and Europe, respectively. The antioxidant and phenolic properties of their water soluble phenolic extracts could be important for human nutritional health. S.c. cultivars (Johns, CSAM72, Adams II, Brush Hill 2, Maxima, Scotia, Eridu 1, and York) and S.n. (Albo‐variegata, Purpurea, Samdal, and Guincho Purple) were cultivated at the USDA ARS National Clonal Germplasm Repository, Corvallis, Oregon. Ripe fruits were harvested, frozen at −80C, and aqueous extracts prepared/pasteurized. Phenolic content (gallic acid equivalence mg/L; GAE) for S.n. (6751 ± 528) was significantly (P=0.004) greater than S.c. (5271 ± 806). Effects of elderberry antioxidants on human LDL oxidation (5μM Cu++) lag‐time (A234nm) was examined at extract dilutions of 1:10,000, 1:50,000 and 1:100,000. At the 1:10,000 dilution, antioxidant activity for S.n. (201±12 min) was significantly higher than S.c. (138±12 min), 1:50,000 and 1:100,000 dilutions were not associated with significant antioxidant activity. Higher GAE was correlated with increased lag‐time (R2= 0.55). Understanding differences in elderberry antioxidant activity may be important for development of elderberry nutritional health applications and cultivar selection.
Quality Characteristics of Korean Traditional Pancake(Jeon) Containing Blueberry for the Global Marketing as a Functional Food
Authors: jung ji-hye , yang jinyoung , park junghyun , shin junghee , kim hyunsook
Published: June 2021

Korean traditional pancake, a popular traditional food, is often consumed as a side dish in Korea. However, it is very calorific and contains a lot of sodium so not ideal for people with overweight. We attempt to develop functional Korean traditional pancake low in calorie, with a good taste for the world. The Korean traditional pancake was made with blueberry(0% BBP, 16.5% BBP, 22.5% BBP). To exam the quality characteristic, nutritive components, total polyphenol and antioxidant activity(2,2‐diphenyl‐1‐picrylhydrazyl( DPPH) radical scavenging activity, Oxygen Radical Absorbance Capacity(ORAC) activity) were measured. The amount of calorie and sodium were decreased in a dose dependent manner(0% BBP; 177.16 kcal/448.93 mg, 22.5% BBP; 122.2 kcal/288.87 mg). Total contents of polyphenol were significantly high in 22.5% BBP(220.05 ± 10.98 mgGAE/100g sample) and the lowest in 0% BBP(67.61 ± 1.61 mgGAE/100g sample). The DPPH activity was the highest value of 62.81 ± 0.74% in the 22.5% BBP. As the results, this study showed that Korean traditional pancake containing blueberry decreased the calorie, sodium and improved antioxidant enzyme activity. This work was supported by Korean Food Foundation and BK 21.

Grant Funding Source: Korean Food Foundation
Flavanols exert cardiovascular health benefits in humans by altering gene transcription levels in white blood cells
Authors: Weseler Antje R. , Ruijters Erik J. B. , Milenkovic Dragan , Haenen Guido R. M. M. , Bast Aalt
Published: June 2021

Epidemiological evidence highlights that a diet rich in flavanols reduces the risk of cardiovascular disease. The molecular mechanisms by which flavanols establish their vascular health effects in humans are still not fully clear.

In a randomized, double‐blind, placebo controlled study gene transcription levels were determined in white blood cells of male smokers before and at the end of the 8‐weeks supplementation of daily 200 mg of monomeric and oligomeric grape‐seed flavanols. Upon approval by the Medical Ethical Committee of the Maastricht University and Academic Hospital Maastricht, The Netherlands, the study was conducted in accordance with the World Medical Association Declaration of Helsinki of 1975 as revised in 2008.

Flavanols significantly altered expression of genes mediating inflammation, redox processes, lipid metabolism and endothelial homeostasis. The beneficial regulation of transcriptional pathways involved in the pathology of cardiovascular diseases is suggested a major subcellular mechanism by which flavanols sustain vascular health in humans.
Inflammation, weight status, and iron status in Mexican children in a randomized controlled iron‐biofortified bean feeding trial
Authors: Luna Sarah , Villalpando Salvador , Shamah Teresa , Boy Erick , Haas Jere D.
Published: June 2021

Overweight is increasing in transition countries, but iron deficiency remains prevalent. Ferritin is a common marker or iron stores; however, in populations with chronic inflammation, ferritin values may be artificially increased in the presence of real iron deficiency. This study examines the relationship between BMI and markers of iron status and inflammation in children age 9–12 (n = 347) using data from a randomized, placebo‐controlled trial of iron biofortified beans in 20 Mexican boarding schools. Dependent variables are iron status (hemoglobin, ferritin, and sTfR) expressed as continuous or categorical variables (iron deficient and anemic). BMI is expressed as a continuous or categorical variable (normal, overweight, obese). Inflammation was assessed by CRP and AGP. Covariates, such as age, sex, school, amount of fortified beans consumed were controlled. Overweight was observed in 17.6% of the children, 13.5% were anemic (hemoglobin <120 g/L), 9.8% had elevated CRP (>3 mg/L) and 11.0% had elevated AGP (>1 g/L). Inflammation status of the school, obesity, and individual baseline CRP predicted elevated ferritin at baseline (p =0.003; p=0.022; p<0.001, respectively). In boarding schools given the control beans, those with high morbidity showed a positive change from baseline in corrected log ferritin (p = 0.0074) compared to those with low morbidity. Supported by HarvestPlus and AgroSalud
A Novel Single Nucleotide Polymorphism within the NOD2 Gene is Associated with Pulmonary Tuberculosis in Chinese
Authors: Zhao Mengyuan , Jiang Feng , Liu Jiyan , Xue Yun , Zhang Wangjiang , DENG Xiling , Wu Fang , Zhang Le , Zhang Xing , Zhang Yuxiang , Li Fujian , Wei Liliang , Fan Dapeng , Sun Xiaojun , Jang Fengfeng , Li Jichen
Published: June 2021

The immune system relies on pattern recognition receptors for the identification of invasion by exogenous pathogenic microorganisms. Nucleotide‐binding oligomerization domain2 is a member of the NLR family and is considered as non‐redundant recognition mechanism of Mycobacterium tuberculosis. The present study was conducted on the fourth exon of the NOD2 gene in 179 cases and 177 healthy controls using PCR sequencing, and found a synonymous mutation Arg587Arg which was located in the ultra‐coil encoding region (Winged Helix and Superhelical Domain) of the NOD2 protein. The synonymous mutation was CGT to CGG encoding the Arginine (OR: 2.06; CI: 1.18–3.60). There was a significant difference between tuberculosis patients and the healthy control group (P<0.05), indicating the Arg587Arg site susceptibility to pulmonary tuberculosis. By focusing on the Pro268Ser, Arg708His and Aal725Val locus in the NOD2 gene, we found that the probability of this series was very low in a Chinese Han population and there was no significant difference (P=0.267). We concluded that the single nucleotide polymorphism Arg587Arg in the NOD2 gene was significantly associated with tuberculosis in the Chinese population that provided new data for the world tuberculosis susceptibility research.
A novel real‐time PCR assay to detect Factor V Leiden and heterozygote Factor V Leiden using HyBeacons probe chemistry
Authors: Parrish Torrey , Duggan Megan , Williams Patrick
Published: June 2021

Factor V Leiden (FVL) is an inherited disorder associated with blood clotting. Approximately 5% of Caucasians have FVL. We have developed a real‐time PCR assay for detection of FVL from buccal swabs or blood samples. A HyBeacons probe is used to detect FVL during melt curve analysis. HyBeacons probes are hybridization probes that allow simultaneous identification of single nucleotide polymorphisms (SNPs) via melt curve analysis. The probe is designed to hybridize to a region around a SNP associated with FVL. This allows the probe to bind to wild type and mutant forms of Factor V (FV). A normal amplification curve is seen for FV and FVL, however, during melt curve analysis a melt peak temperature shift is observed between FV and FVL. To optimize the assay, plasmids were constructed to both FV and FVL sequences. The assay was then run against each individual plasmid and a mixture of both to determine melt peaks for FV, FVL, and heterozygous samples. The melts peaks were approximately 52.3°C for FV, 57.5°C for FVL, and heterozygous samples had peaks at 52.3°C and 57.5°C. Twenty blood and buccal samples were obtained. DNA was extracted from the samples using Evogen ONE‐B and Evogen ONE, respectively and used in the FVL assay. All samples showed amplification but were negative for FVL. This newly developed assay is able to detect FV, FVL, and heterozygote from blood and buccal samples using a single HyBeacons probe.
Investigation of anti‐oral cancer molecular mechanism of pipoxolan in oral squamous cell carcinoma cell line
Authors: LI Pei-Chuan , Chou Pei-Yu , Wu Chieh-Hsi , Sheu Ming-Jyh
Published: June 2021

The aim of the present study was to investigate the molecular mechanisms by which pipoxolan exerts inhibitory effects and apoptotic activity in human oral cancer cells. Pipoxolan reduced the cell viability and induced apoptosis in oral squamous cell carcinoma (OSCC) cell line HSC‐3. The effects of pipoxolan on the proliferation of cancer cells and on the distribution of cells within different phases of the cell cycle were investigated using a MTT assay and a flow cytometer, respectively. The effects of pipoxolan on the apoptosis of cancer cells was investigated using Annexin/PI double stain and flow cytometer. Factors affecting the cell cycle and apoptosis, including p53, p21, Bax, Bcl2, cytochrome c caspase 3, caspase 8 and caspase 9 expressions, were examined by Western blotting. Our results demonstrate that pipoxolan significantly induce apoptosis after 24 h exposure in human oral cancer HSC‐3 cells. In addition, HSC‐3 cells were arrested in the G0/G1 phase through the induction of p53/p21 by pipoxolan. Apoptosis was associated with an increased Bax/Bcl‐2 ratio, cytochrome c release, cleavage of procaspases‐9 and ‐3, and hydrolysis of PARP. Pipoxolan also suppressed the expressions of PI3K and phosphorylation of Akt. This is the first report confirming the anti‐cancer activity of pipoxolan against human OSCC HSC‐3.
The organosulfides diallyl sulfide, diallyl disulfide, and diallyl trisulfide accelerate benzo(a)pyrene metabolism in MCF‐10A cells
Authors: Moyler Candace C. , Nkrumah-Elie Yasmeen , Dennis William , Hudson Alicia , Hammamieh Rasha , Darling-Reed Selina
Published: June 2021

Benzo(a)pyrene (BaP) is a naturally and anthropogenically produced carcinogen created during the incomplete combustion of organic material. BaP is considered a dietary, occupational, and environmental carcinogen, and BaP‐associated adducts have been found in the breast tissue and milk of women. Garlic has been shown to have chemopreventive and chemotherapeutic potential, and its organosulfide compounds (OSCs) have been attributed to its anti‐carcinogenic potential. BaP is a procarcinogen, requiring biotransformation to form its highest carcinogenic potential. Normal breast epithelial (MCF‐10A) cells were treated with BaP alone; or were pretreated with either one of the the garlic OSCs diallyl sulfide, diallyl disulfide, or diallyl trisulfide at either 6, 60 or 600 ìM for 4 hrs followed by treatment with 1 ìM BaP. The media from the treatments was extracted after 12 and 24 hrs and analyzed for changes in BaP concentration using high pressure liquid chromatography. All OSCs significantly reduced the concentration of BaP in the media at 12 and 24 hrs; most likely through enhanced biotransformation by the MCF‐10A cells, which converts BaP to metabolites and intermediates. Further studies evaluating the presence of BaP metabolites in the media will determine the significance of OSC–induced BaP metabolism in MCF‐10A cells.
Control of Lipolysis by Perilipins
Authors: Brasaemle Dawn L.
Published: June 2021

The perilipins are a family of 5 proteins that are among the most abundant lipid droplet‐associated proteins in animal cells; most cells in the body express 2 – 4 perilipins. Perilipins form organizing scaffolds at the surfaces of lipid droplets to coordinate the activity of lipases and other factors. Different cells express different combinations of lipases; hence, lipolysis is controlled by a unique combination of perilipins and lipases in a cell‐specific manner. In adipocytes, perilipin 1 serves a critical function in the control of lipolysis catalyzed by hormone‐sensitive lipase (HSL) and adipose triglyceride lipase (ATGL). Under basal (fed) conditions, perilipin 1 restricts the access of lipases to lipid droplets, thus promoting the storage of triacylglycerols. Following binding of catecholamines to cell surface receptors of adipocytes, phosphorylation of perilipin 1 by protein kinase A promotes lipolysis through multiple mechanisms including facilitating recruitment and binding of HSL to lipid droplets and release of CGI‐58, a co‐activator of ATGL, from the perilipin scaffold. Perilipin 5 is highly expressed in oxidative tissues including heart and skeletal muscle, where ATGL and HSL catalyze lipolysis. Perilipin 5 binds both CGI‐58 and ATGL at lipid droplet surfaces, and serves as a negative regulator of lipolysis under basal conditions. Supported by NIH R01 DK54797.
Effects of 3,4‐methylenedioxymethamphetamine (MDMA) and its metabolites on cardiovascular function in rats
Authors: Schindler Charles , Baumann Michael H , Thorndike Eric B. , Blough Bruce E. , Goldberg Steven R.
Published: June 2021

The abuse of MDMA (Ecstasy) peaked in the early 2000s, yet reports of MDMA use from emergency room admission data continue to increase. The potential for cardiovascular effects of MDMA to contribute to its acute toxicity has long been recognized. However, the role of MDMA metabolites in mediating the drug's cardiovascular actions has not been studied. Here we employed radiotelemetry to evaluate the effects of racemic MDMA and its metabolites on blood pressure (BP), heart rate (HR) and locomotor activity in conscious rats. MDMA (1–20 mg/kg, s.c.) increased BP, HR, and activity. The rise in HR peaked at lower doses than the effects on BP and activity. The MDMA metabolite 3,4‐methylenedioxyamphetamine (MDA) produced effects that were similar to MDMA. The metabolite 3,4‐ dihydroxymethamphetamine (1–10 mg/kg, HHMA) increased HR to a greater degree than MDMA. The metabolite 3,4‐ dihydroxyamphetamine increased HR, but less than HHMA. These dihydroxy metabolites did not alter motor activity. The metabolites 4‐hydroxy‐3‐methoxymethamphetamine and 4‐ hydroxy‐3‐methoxyamphetamine (3–10 mg/kg) did not affect BP, HR or activity. The tachycardia produced by MDMA and HHMA may be mediated via beta‐adrenergic receptors as propranolol attenuated the HR increase. Our results suggest that HHMA may contribute significantly to the overall cardiovascular effects of MDMA in vivo. Supported by NIH/NIDA Intramural Research Funds.
Effects of Combinations of Metyrapone and Oxazepam on Methamphetamine Cue Reactivity and the HPA Axis in Rats
Authors: Miller Andrew Joseph , Cornett Elyse , Guerin Glenn , Goeders Nick
Published: June 2021

METH is a highly addictive psychostimulant. There is currently no FDA approved treatment for METH addiction. Study in our lab focuses on the role of the stress response and the influence of environmental cues in accounting for the high rate of relapse in recovering addicts (60% relapse for METH patients).

The combination of Metyrapone (MET, a CORT synthesis inhibitor) and Oxazepam (OX, a benzodiazepine) has been shown to decrease both cocaine self‐administration and cue‐induced reinstatement. The MET/OX combination was used in these experiments to test its effects on cue‐induced METH craving in rats.

Rats were trained to self‐administer METH via lever response. Drug delivery was paired with a tone and the illumination of a houselight, both serving as secondary reinforcers, or cues. After stable self‐administration of METH for at least 10 days, subjects entered a 2‐week abstinence period. Rats were then given a cue reactivity test wherein the cues were presented at lever response, but no METH was delivered. With vehicle pretreatment, the response‐contingent presentation of the cues reliably maintained METH seeking (i.e. lever responses). Pretreatment with the MET/OX combination attenuated responding at both doses tested, suggesting diminished craving. Levels of stress hormones corticosterone (CORT) and ACTH were measured to ensure that the MET/OX combination did not cause adrenal insufficiency.
Sigma receptor ligands attenuate methamphetamine‐induced hyperthermia but do not modulate IL‐1B mRNA expression in select brain regions
Authors: Seminerio Michael John , Robson Matthew J , Mesangeau Christophe , Narayanan Sanju , McCurdy Christopher R , Matsumoto Rae R
Published: June 2021

Methamphetamine is one of the most widely abused drugs worldwide, with hyperthermia being a primary cause of death. Methamphetamine‐induced hyperthermia involves a variety of cellular mechanisms, including increases in IL‐1β. Previous studies indicate methamphetamine interacts with sigma receptors and selective sigma receptor ligands attenuate some of the effects of methamphetamine, including hyperthermia. The purpose of the current study was to determine if sigma receptor ligands can attenuate acute increases in body temperature following methamphetamine administration and whether these protective effects occur through modulation of IL‐1β. To evaluate the role of sigma receptors, male, Swiss Webster mice were pretreated with AZ66 or SN79 15 min prior to an acute bolus dose of methamphetamine and body temperatures were recorded for up to 45 min. Acute, bolus administration of methamphetamine caused a significant, dose dependent increase in body temperature, and pretreatment with each of the sigma receptor ligands significantly attenuated methamphetamine‐induced hyperthermia. To determine if these protective effects were related to the modulation of IL‐1β, the mouse striatum, cortex and hypothalamus were collected 45 min after drug administration and evaluated for IL‐1β mRNA expression. Methamphetamine produced a significant increase in IL‐1β mRNA in each of these brain regions; however, sigma receptor ligands failed to attenuate the changes in IL‐1β mRNA expression. It appears that sigma ligands can mitigate methamphetamine‐induced hyperthermia through a different mechanism from that involved in the modulation of IL‐1β mRNA expression. (Supported by NIDA DA013978, DA023205)
Chronic α1AAR Stimulation May Increase Adult Neurogenesis and Parvalbumin Interneurons
Authors: Nielsen Amber Rose Dybdal , Collette Katie Marie , Perez Dianne , Doze Van Alan
Published: June 2021

Adult neurogenesis occurs in the subventricular zone (SVZ) of the lateral ventricles and the subgranular zone (SGZ) of the dentate gyrus. Adult neurogenesis seems to play a role in depression, anxiety, cognition, epilepsy, and schizophrenia with increases having a positive effect. Chronic stimulation of alpha‐1A adrenergic receptors (α1AARs) has been shown to increase adult neurogenesis in the SVZ; however, its effects have not yet been thoroughly explored in the SGZ. Persons suffering from schizophrenia have decreased numbers of parvalbumin‐positive (PV+) interneurons. Preliminary work in our lab has shown an increase in PV+ interneurons after chronic activation of the α1AAR. The present study investigated the impact of chronic α1AAR stimulation on neurogenesis and on PV+interneuron numbers in the adult SGZ. Constitutively active mutant (CAM) α1AAR mice, which have a chronically activated α1AAR, were used and compared with appropriate control animals. The mice were cardiac perfused, their brains sectioned, and double‐labeled immunohistochemically with 5‐bromo‐2‐deoxyuridine (BrdU) and parvalbumin antibodies. The results indicate a possible increase in BrdU+ and PV+ interneurons. Treatments to increase adult neurogenesis may be used to treat depression and anxiety in the future. Similarly, treatments to increase adult neurogenesis and PV+interneurons may improve symptoms of schizophrenia.
Neuronal Nicotinic Acetylcholine Receptors (nAChRs) Are Involved in Nicotine‐Mediated Reversal of Ataxia in an Animal Model
Authors: Lambert Chase Steven , Philpot Rex M , Engberg Melanie E , Wecker Lynn
Published: June 2021

Clinical data indicate that the neuronal nicotinic acetylcholine receptor (nAChR) partial agonist varenicline ameliorates abnormalities in gait and balance in patients with spinocerebellar ataxia type 3, but the mechanisms involved have not been investigated. The goal of this study was to determine whether nicotine (Nic) could improve gait in an animal model of ataxia and whether this effect was mediated by an action at nAChRs. Following baseline assessment of locomotor activity and gait, male Sprague‐Dawley rats received injections of the neurotoxin 3‐acetylpyridine (3‐AP) to preferentially destroy the olivocerebellar pathway, and behaviors were reassessed one week later. Subsequently, animals received daily injections of either saline or mecamylamine (Mec, 1 mg/kg), followed 30 minutes later by either saline or Nic (1 mg/kg) for 1 week. The administration of 3‐ AP led to a 21–23% decrease in distance moved and velocity of movement, an 8% decrease in hindpaw stride length and a 25% increase stride width. Nic administration for 1 week did not alter locomotor activity or velocity, but did improve hindpaw stride length and width, effects that were prevented by prior Mec administration. Results indicate that activation of nAChRs is of benefit in an animal model of olivocerebellar degeneration. Current experiments are determining the specific nAChRs involved and the cellular mechanism mediating this effect. Supported by NIH #NS072114‐01
Renal drug transporter expression in pregnant mice with type 1 diabetes
Authors: Yacovino Lindsay , Aleksunes Lauren
Published: June 2021

Sex hormones and metabolic changes (such as obesity and hyperglycemia) can alter renal transporter expression in rodents. The purpose of this study was to investigate renal transporter expression in response to pregnancy and type 1 diabetes. Female mice were treated with vehicle or the diabetogenic drug, streptozotocin (STZ) 2 weeks prior to mating. Renal transporter mRNA and protein expression were evaluated in vehicle‐ and STZ‐treated pregnant and virgin mice. The majority of renal transporters were unaffected by pregnancy in vehicle‐treated mice, with only modest 40–80% increases in Mrp3, Oatp2b1, Pept1, and Cnt1 mRNA. In contrast, renal transporters were markedly changed in STZ‐treated virgin mice with decreased Oat2, 5, and Mrp3 mRNA and elevated Oatp2b1, 4c1, Mrp2, 4, 5, and Pept1 mRNA. In combination, STZ treatment and pregnancy downregulated Oatp4c1, Mdr1b, and Mrp3 mRNA. Transcriptional profiling suggests Hnf1α, Nrf2, PXR, AhR, and PPARα may be involved in regulating renal transporter expression. In conclusion, renal drug transport pathways are altered by hormone changes that occur during pregnancy and diabetes. Supported by ES‐007148, ES‐005022, DK‐080774, ES‐020522.
Evaluation of aminoglycosides‐induced cardiotoxicity in zebrafish obtained in a low cost facility
Authors: Vargas Rafael Antonio
Published: June 2021

Zebrafish is a popular model in basic medical sciences research since middle eighties. Cardiovascular research has been made in larvae and adults and different aspects of cardiovascular development and physiology has been described. Zebrafish heart is sensible to external chemicals agents and cardiac alterations in shape and function could be observed under conventional microscopy.

Currently, we describe the set up of a low cost, custom‐built facility to produce zebrafish. In addition, we explore toxicity of aminoglycosides on cardiac activity of transparent zebrafish larvae. We have found that some aminoglycosides can cause arrhythmic activity at high doses. This effect has some similarities to the arrhythmogenic effect caused by classical antiarrhythmic drugs.

Use of zebrafish as a model to investigate pharmacological and toxicological properties of different drugs could be an inexpensive, an ideal model, which could be an advantage for researchers in undeveloped countries where to obtain financial aid, in many cases, is too complicated.
Nitric Oxide Synthase, But Not Heme Oxygenase, Mediates The Adenosine A2B Receptor‐Sensitive Renal Vasodilations In Female Rats
Authors: El-Mas Mahmoud M , El-Gowelli Hanan M , Elsalakawy Lamiaa K , El-gowilly Sahar M
Published: June 2021

Stimulation of adenosine receptors by 5′‐N‐ethylcarboxamidoadenosine (NECA) dilates the renal vasculature. Studies were conducted in phenylephrine‐preconstricted isolated perfused kidneys of female rats to determine: (i) the relative contributions of adenosine receptor subtypes to NECA responses and (ii) the potential modulation of these responses by nitric oxide synthase (NOS) and/or heme oxygenase (HO). Cumulative bolus injections of NECA (1.6–50 nmol) elicited dose‐related vasodilations. NECA responses were attenuated after blockade of A2B (alloxazine) but not A2A [8‐(3‐Chlorostyryl) caffeine] or A3 (VUF 5574) receptors, suggesting preferential involvement of A2B receptors in NECA vasodilations. NOS activation appears to mediate the A2B‐dependent NECA response because: (i) NOS inhibition by L‐NAME attenuated NECA responses, (ii) no additional inhibition of NECA responses occurred upon concurrent exposure to L‐NAME and alloxazine, and (iii) L‐arginine abrogated the alloxazine‐evoked inhibition of NECA responses. Conversely, HO inhibition by zinc protoporphyrin affected neither NECA responses nor the attenuating effect of alloxazine on these responses. Overall, the activation of A2B/NOS signaling underlies the vasodilatory effect of NECA in female kidneys.

This project was supported financially by the Science and Technology Development Fund (STDF), Egypt, Grant No. 502.
Maternal nutrient restriction during pregnancy impairs an EDHF‐like pathway in fetal coronary arteries without affecting large conductance, calcium‐activated K channel (BKCa) activity in smooth muscle cells
Authors: Shukla Praveen , Modgil Amit , Reynolds Larry , Caton Joel , Sun Chengwen , O'Rourke Stephen
Published: June 2021

We demonstrated previously that maternal nutrient restriction during pregnancy impairs an EDHF‐like pathway that is mediated via BKCa activation in fetal coronary arteries (CA). We now show that the impairment is not likely due to altered BKCa activity in CA smooth muscle cells. Pregnant ewes were fed a control or nutrient restricted diet from day 50–130 of gestation. In CA isolated from control fetal lambs, relaxation to bradykinin was partially inhibited by nitro‐l‐arginine (NLA). Iberiotoxin nearly abolished the NLA‐resistant response to bradykinin. In CA from undernourished animals, relaxation to bradykinin was fully suppressed by NLA. Whole‐cell BKCa currents were nearly identical in CA smooth muscle cells from control and nutrient restricted animals (peak current density= 46 ± 5 vs. 39 ± 6 pA/pF; n=6). 14,15‐EET (putative EDHF in CA) caused fetal CA relaxation and BKCa activation that was unaffected by maternal nutrient restriction. Similar results were obtained with the BKCa‐openers, BMS 191011 and NS 1619. Relaxation to bradykinin in fetal CA is mediated by NO, and by an EDHF‐like pathway that involves BKCa activation. Maternal undernutrition during pregnancy results in loss of the EDHF‐like pathway in fetal CA, an effect that is likely due to endothelial dysfunction (i.e. impaired synthesis and/or release of an EDHF‐like mediator) rather than impaired BKCa activity in CA smooth muscle cells. (NIH HD61532)
Propofol regulation of vascular reactivity is mediated via TRP channels
Authors: Sinha Sayantani , Kmetz John G. , Damron Derek S. , Bratz Ian N.
Published: June 2021

Transient receptor potential channels, TRPA1 and TRPV1 are known to play a major role in pain and inflammatory pathways. Recent evidences have shown that general anesthetics can either directly or indirectly sensitize these channels. Similarly studies have also implicated cross‐talk between these channels which plays an immense role in regulating these channels. Propofol, an intravenous anesthetic, causes hypotension when administered to patients during surgery. Our main objective was to determine the role of these channels in the regulation of vascular reactivity while further demonstrating Propofol mediates vasodilation through TRPA1/TRPV1 signaling. Using high‐fidelity microtip transducer catheter, changes in mean arterial pressure were recorded in C57BL6 and TRPA1(−/−) mice. Propofol induced a dose‐dependent depressor response which was blunted in TRPA1(−/−) mice and was further attenuated in TRPA1(−/−) mice in the presence of a specific TRPV1 antagonist SB366791. Isometric‐tension studies in endothelium intact and denuded aortic rings from TRPA1(−/−)mice in presence/absence of SB366791 illustrated a role for TRPA1/TRPV1. Furthermore propofol elicited robust relaxation in isolated coronary microvessel from controls which was blunted in the presence of SB366791 and in vessels from TRPA1(−/−)mice. In conclusion, Propofol mediated regulation of vascular reactivity occurs in part via combined TRPA1/TRPV1 mediated signaling.
Role of TRPC3 channels in BDNF‐induced plasticity, hippocampal neuronal excitability and memory
Authors: Tryba Andrew Kieran , Chong Jayhong A. , Del Camino D. , Lacey V. , Abramowitz Joel , Birnbaumer Lutz , Harder David R. , Gerges Nashaat , Kaczorowski Catherine C.
Published: June 2021

Brain‐derived neurotrophic factor (BDNF) enhances synaptic plasticity, which is a candidate mechanism for improving hippocampus‐dependent memory. Here, we demonstrate that BDNF also modulates intrinsic membrane conductances associated with improved memory function. In doing so, BDNF reduces the intrinsic excitability of regular firing (RF) neurons, yet increases the intrinsic bursting properties of burst firing (BF) neurons in the subiculum ‐ the primary efferent pathway of hippocampus that plays a role in memory, drug abuse relapse and temporal lobe seizures. BDNF reduces RF excitability by activating transient receptor canonical potential (TRPC3) channels, which enhances the fast afterhyperpolarization (fAHP). In contrast, BDNF‐enhances intrinsic bursting of BF neurons via a TRPC3‐ independent pathway. Blocking TRPC3 channels suppresses BF activity yet promotes bursting behavior in RF neurons, likely by suppressing AHP. Manipulations that promote bursting and/or suppress the AHP have been suggested to improve memory function. Here, we show that TRPC3 knockout (KO) mice have enhanced hippocampal‐dependent conditioned fear memory. As BDNF is enhanced during learning, BDNF signaling in TRPC3 knockout mice likely promotes bursting, which may explain the improved memory in TRPC3 KO mice. Modulating TRPC3 channels may provide a novel treatment for memory disorders in aging and Alzheimer's Disease.
AMP‐activated protein kinase activation is associated with an inhibition of fibrotic properties of cardiac fibroblasts
Authors: Horman Sandrine , Noppe Gauthier-Thibaut , Buchlin Patricia , Marquet Nicolas , Baeyens Nicolas , Morel Nicole , Vanoverschelde Jean-Louis , Bertrand Luc , Beauloye Christophe
Published: June 2021

Purpose

We asked whether cardiac fibroblast (CF) function might be regulated by AMP‐activated protein kinase (AMPK). We also investigated cardiac fibrosis in AMPK knockout mice (KO) after myocardial infarction.

Methods

CF were treated by Transforming growth factor‐beta‐1 (TGF‐beta‐1), serum or platelet derived growth factor (PDGF‐BB) to stimulate myodifferentiation, proliferation and migration respectively. AMPK was activated prior to the different treatments with thienopyridone derivative A‐769662. For in vivo studies, KO mice and their wild‐type (WT) littermate were submitted to a ligation of the coronary artery to mimic myocardial infarction. Gene expression in the left ventricule was measured by qRT‐PCR.

Results

A‐769662 compound dose‐dependently prevented TGF‐beta‐ 1‐induced myodifferentiation. In addition, it decreased by 50% proliferation and completely prevented PDGF‐induced migration of CF, without affecting their mortality. Studies in KO mice show that RNA expression for collagen I and III was significantly increased in the infarct area of KO mice compared to WT mice, after myocardial infarction.

Conclusion

Our data indicate that AMPK could be a negative regulator of CF function, and by this way decrease the fibrotic component of ventricular remodelling which takes place after myocardial infarction.
Distinct atrial and ventricular microRNA changes in experimental heart failure
Authors: Chen Yu , Wakili Reza , Luo Xiaobin , Naud Patrice , Kaab Stefan , Dobrev Dobromir , Nattel Stanley
Published: June 2021

Heart failure (HF) causes different forms of remodeling in left atrium (LA) vs left ventricle (LV), with fibrosis much more prominent in LA. MicroRNAs (miRs) are central regulators in cardiac remodeling. This study compared LA and LV miRNA changes during the development of experimental HF.

Methods

HF was induced in dogs by ventricular tachypacing (VTP, 240 bpm) for 0 (CTL), 12, 24 hrs, 1, 2 and 5 wks (n=5–8/group). MiR expression was assessed by microarray and qPCR.

Results

Microarrays showed greater changes in LA vs LV (Fig A). MiR‐29b, miR‐133a and miR‐30a decreased within 24 hrs; MiR‐21 and miR‐146b increases started at 1 wk only in LA (B, C). In contrast, miR‐214 and miR‐146a showed similar changes in LA and LV (D). MiR‐21, miR‐29b, miR‐133 and miR‐30 are implicated in extracellular‐matrix (ECM) remodeling. Collagen‐1/ ‐3, fibronectin 1 and fibrillin increased ~3.8–26‐fold*** to steady state at 1–2 wk, TIMP‐3 decreased from 24 hrs (~58%***) while MMP‐9 increased at 12 hrs (~9‐fold***) and remained elevated. Changes were much smaller in LV at all times, agreeing with atrial‐predominant fibrosis. miR‐214 and miR‐146 target IKs and ICa,L subunits, known to change in HF LA and LV.

Conclusions

VTP‐induced HF causes greater miRNA changes in LA vs LV. LA expression of miRs with effects on ECM synthesis, paralleled by related ECM expression changes, implicate miRNAs in HF related LA‐selective structural remodeling.
Myoendothelial Junction Formation is Restricted in Pulmonary Arteries of Fetal Sheep
Authors: Lumbard Britney , Waskel Edith , Paez Ricardo , Blood Quintin , Rubalcava Monica , Kim Joon H , Longo Lawerence D , Wilson Sean M
Published: June 2021

Endothelial cell activation can hyperpolarize vascular myocytes, resulting in myocyte relaxation and vessel dilation. Direct communication by way of myoendothelial junctions (MEJ) made through the internal elastic lamina (IEL) contributes to vasodilatory responses. The function of the MEJ in pulmonary arteries is not completely understood and less is known regarding their function before birth. The fetal lung has little blood flow and endothelium‐dependent relaxation of pulmonary arteries is limited, which lead us to hypothesize that MEJs are underdeveloped in the fetal lung. This hypothesis was tested using laser scanning confocal microscopy techniques and viewing the IEL of pulmonary arteries from fetal and adult sheep. The IEL was viewed using autofluorescence, and MEJ's were visualized using cell mask red fluorescence. The number and size of pores in the IEL were quantified to index potential MEJ sites and this was compared to the number of MEJ sites that stained positive for cell membrane. Preliminary data suggests the size of each IEL pore, total pore area, and percentage of IEL holes with MEJs was greater in the adult. The restricted IEL pore and MEJ formation in the fetus could diminish MEJ –dependent myocyte hyperpolarization and resultant vasodilation. This could contribute to the high vascular resistance in the fetal pulmonary vasculature. Support from HHMI, NIH and NSF.
Rap1 and afadin promote interactions between adherens junctions and tight junctions and regulate endothelial permeability
Authors: Birukova Anna A , Zebda Noureddine , Wu Tinghuai , Dubrovskyi Oleksii , Sarich Nicolene , Birukov Konstantin G
Published: June 2021

Oxidized 1‐palmitoyl‐2‐arachidonoyl‐sn‐glycero‐3‐phosphocholine (OxPAPC) enhances vascular endothelial barrier properties in vitro and in vivo. This study examined the role of small GTPase Rap1 and its downstream target afadin in regulation of OxPAPC‐induced interactions between adherens junctions and tight junctions, the major gatekeepers of endothelial barrier. Immunofluorescence analysis, subcellular fractionation, and co‐immunoprecipitation assays revealed OxPAPC‐induced translocation of cell junction adaptor protein afadin to the cell periphery. OxPAPC also promoted accumulation of adherens junction proteins: VE‐cadherin, p120‐catenin, and beta‐catenin and tight junction proteins: ZO‐1, occludin, and JAM‐A in the cell membrane. OxPAPC induced novel cross‐interactions between adherens junction and tight junction protein complexes, which were dependent on the OxPAPC‐induced Rap1/afadin activation. Inhibition of Rap1 or afadin function suppressed OxPAPC‐mediated endothelial barrier enhancement as well as it inhibited protective effects of OxPAPC against ventilator‐induced lung injury in vivo. These results show the critical role of Rap1/afadin‐mediated interactions between adherens junction and tight junction complexes in the pulmonary vascular endothelial barrier preservation.

Funding: NHLBI HL089257, HL107920, HL76259, HL58064, HL89257