bluuebunny/crossref_metadata_2025 huggingface dataset:
A high fiber cookie made with VERSAFIBE&[trade] 1490 dietary fiber reduces post‐prandial glucose and insulin responses in healthy adults
Authors: Stewart Maria L , Zimmer J. Paul
Published: November 2023

VERSAFIBE™ 1490 dietary fiber is a resistant starch type 4 derived from potato, containing 90% total dietary fiber (TDF, AOAC 991.43 method). This low‐viscosity fiber ingredient has many food applications, including snacks, bread, pasta, and cookies. In this double‐blind, randomized, cross‐over study, we compared the glycemic response to two cookies in 28 healthy adults. The test cookie was formulated with VERSAFIBE 1490 dietary fiber and refined wheat flour (24.1 g TDF) and the control cookie was formulated with maltodextrin and refined wheat flour (0.5 g TDF, control). The cookies were matched for total weight, total carbohydrate, sugars, protein, and fat. Subjects were randomly assigned the treatment order. The primary outcome was the two‐hour (baseline to 120 min) intravenous blood glucose incremental area under the curve (iAUC0‐120min). During each 24‐hour study period, subjects consumed a standard evening meal, fasted for 12 hours, and arrived at the study clinic the following morning. Serum glucose, serum insulin, and capillary glucose were measured at 0 min, 15 min, 30 min, 45 min, 60 min, 90 min, 120 min, 180 min and 240 min after cookie consumption. This study was conducted in accordance with the ethical principles outlined in the Declaration of Helsinki and approved by the Institutional Review Board (IRB Services, Aurora, Ontario). All subjects provided informed consent prior to enrolling in the study. Product perception (sensory evaluation and overall liking) was similar between the two treatments. The VERSAFIBE 1490 dietary fiber cookie reduced the post‐prandial serum glucose iAUC0‐120min by 44% (p = 0.004) and reduced the maximum glucose concentration (Cmax0‐120min) by 8% (p = 0.001) versus the control cookie. Consumption of the VERSAFIBE 1490 dietary fiber cookie resulted in a significant 46% reduction of the post‐prandial serum insulin iAUC0‐120min (p < 0.001) and a 23% reduction in Cmax0‐120min (p = 0.007) versus the control cookie. There was a 48% reduction in the elevation of post‐prandial capillary glucose iAUC0‐120min (p < 0.001) and a 9% decrease in Cmax 0‐120min (p = 0.005) when participants consumed the VERSAFIBE 1490 dietary fiber cookie versus the control cookie. Serum glucose, serum insulin, and capillary glucose values were significantly lower at individual time points when the VERSAFIBE 1490 dietary fiber cookie was consumed versus the control cookie. This study shows that VERSAFIBE 1490 can be incorporated into a cookie and significantly reduce post‐prandial glucose and insulin responses in healthy adults. The consumer perception of the high‐fiber VERSAFIBE 1490 dietary fiber cookie was similar to the low‐fiber control cookie, which demonstrates a practical approach to formulating baked goods with high dietary fiber content.

Support or Funding Information

This study was funded by Ingredion Incorporated, Bridgewater, NJ, USA. Copyright 2016 by Ingredion Incorporated
Effect of Ganjang (Korean fermented Soy sauce) on renin‐angiotensin‐aldosterone system in Sprague‐Dawley rats
Authors: Kim Misun , Mun Eun‐Gyung , Cha Youn‐Soo
Published: November 2023

This study investigated the effect of Ganjang (Fermented soy sauce) on renin‐angiotensin‐aldosterone system(RAAS) in Sprague‐Dawley rats. Animals were divided into 5 groups; Control(CT), NaCl(NC), Soy bean(SB), Meju(MJ), Ganjang(GJ). The order of Systolic blood pressure(SBP) among the groups were NC>>MJ>CT>GJ>SB. Na+ concentration in the groups were as follows CT<GJ<NC=SB<MJ in urine. In feces, Na+ was not significantly different. Ion excretion indirectly show activation of RAAS which was higher in CT compared to NC, SB, MJ and GJ. Plasma Renin level were as follows CT=NC>GJ>MJ=SB. However, plasma Angiotensin II level and aldosterone level were not significantly different. RAAS is activated by low salt concentration in blood or decreased blood flow. Plasma renin level of NC group was higher and SBP of NC group was increased; SBP of SB, MJ and GJ were decreased. Plasma Angiotensin II and aldosterone levels were not significantly different, although SBP in SB, MJ and GJ groups were decreased. However, mRNA expression of Renin expression was CT>NC=SB=MJ>GJ and Angiotensin I converting enzyme (ACE) expression order was MJ>SB>NC=GJ=CT. Mineralocorticoid receptor (MR) expression was significantly low in GJ compared to other groups. Na+ transporters expression showed similar results. And SB was lower in Na+/K+ ATPaseα1(NKAα1) and Na+/H+exchanger3 (NHE3). MJ showed lower expression of Na+/HCO3− (Bicarbonate) coexchanger (NBC) and Carbonic anhydrases II (CAII) and NHE3. MR is aldosterone receptor and its activation causes in reabsorption of sodium. Therefore, the decreased SBP in SB, MJ and GJ could be a result of low of Na transporters expression. In conclusion, this study showed that the probability of hypertension is lower in Soy sauce including diet compared to the high salt, evident by low level of SBP and sodium transporters in SB, MJ and GJ group compare to NC group.

Support or Funding Information

[This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education(2015R1D1A3A01015869)]
An updated approach for extraction and detection of human plasma tocopherols and tocotrienols
Authors: Khan Md Shahjalal Hossain , Hao Lei , Shen Chwan‐Li , Wang Shu
Published: November 2023

Introduction

Tocopherols and tocotrienols are two subgroups of Vitamin E with a saturated or unsaturated side chain, respectively. Each subgroup has four isomers (α, β, γ and δ). Among four tocopherols and four tocotrienols, α‐ and γ‐tocopherol are the main form of vitamin E found in daily food. Tocotrienols are not abundant in foods, but some studies have demonstrated that they have beneficial effects on health via their anti‐inflammatory and anti‐oxidative properties. A 12‐week randomized, double‐blinded, and placebo‐controlled clinical trial has been conducted to evaluate the effects of γ‐ and δ‐tocotrienols on bone health and their underlying mechanisms. Accurate and reliable extraction and detection methods are critically needed for measuring blood concentrations of tocopherols and tocotrienols in this study.

Methods

Plasma was saponified by KOH followed by extraction with hexane. Rac‐tocol was used as an internal standard. The extracted lipids were dried under nitrogen gas. The dried lipids was resuspended into methanol and transferred into a high performance liquid chromatography (HPLC) vial. Tocopherols and tocotrienols were detected using a Waters HPLC system with a PFP column (4 μm, 4.6×150mm) and a florescence detector, excitation wavelength was 296 nm, and emission wavelength was 325 nm. The mobile phase was composed of methanol and water (35:9, v:v), the flow rate was 0.8 mL/min.

Results

Rac‐tocol, α‐, β‐, γ‐, δ‐tocopherols and α‐, β‐, γ‐, δ‐tocotrienols were separated very well using this HPLC detection method. Compared to the baseline levels, 12‐week supplementation of γ‐ and δ‐tocotrienols increased their blood concentrations.

Conclusions

The extraction method with saponification can effectively extract plasma tocopherols and tocotrienols. The HPLC detection method can simultaneously measure internal standard and all tocopherols and tocotrienols.
Bioavailability of Carotenoids and Specific Vitamins of a Commercially Available Fruit‐, Berry‐ and Vegetable‐Juice Concentrate
Authors: Dams Sebastian , Lamprecht Manfred , Holasek Sandra Johanna , Meier‐Allard Nathalie , Poncza Brigitte , Jansenberger Yvonne , Malliga Daniela Eugenia
Published: November 2023

Introduction

Several clinical trials have documented that supplementations with a whole food based encapsulated fruit/berry/vegetable juice powder supplement (Juice Plus+®) improved markers of redox biology, low‐grade inflammation and circulation. The aim of this study was whether this product provides bioavailable carotinoides and vitamins A, E and C to support mechanistic explanations for the beneficial outcomes observed in the past.

Methods

In this one‐arm clinical trial 18 healthy subjects (m/f, 20 – 50 yrs), non‐smokers, who fulfilled the inclusion criteria received the product over 8 weeks. 6 capsules a day, 2 of the fruit blend + 2 of the berry blend + 2 of the vegetable blend, were taken with meals. After wash‐out of all food supplements and dietetic foodstuff the bioavailability of several carotenoids and vitamins C, A, E was determined at baseline and after 8 weeks of intervention with the nutraceutical.

Results

After 8 weeks of supplementation the plasma carotenoids α‐carotene, β‐cryptoxanthine, and lycopene increased significantly (p<0.05). β‐carotene increase almost reached significance (p=0.053). Also plasma concentrations of vitamins C, A (all‐trans retinol), and α‐tocopherol increased significantly after eight weeks of supplementation. We observed no differences between genders. For those micronutrients with known reference ranges like α‐carotene, β‐carotene, and vitamins C, E and A, all observed increases leveled off around the upper limit of the individual reference range.

Conclusion

The data demonstrate that the investigated fruit‐, berry‐ and vegetable‐juice concentrate is able to increase plasma carotenoids and vitamin concentrations of healthy subjects within 8 weeks. Our observations let us assume that carotenoid‐ and vitamin‐malnourished subjects could benefit from supplementation with this whole food based nutraceutical.

Support or Funding Information

This study was funded by Green Beat, Graz, Austria
GENE EXPRESSION AND COORDINATION OF CELLULAR ZINC TRANSPORTERS AND METALLOTHIONEINS ARE ALTERED IN TYPE 2 DIABETES MELLITUS
Authors: Samman Samir , Chu Anna , Foster Meika , Hancock Dale P , Petocz Peter
Published: November 2023

Background/Aims

The involvement of zinc in multiple physiological systems requires tight control of cellular zinc concentration. The current study aims to explore the relationships among cellular zinc transporters (ZnT and ZIP) and metallothioneins (MT) in a healthy population and a cohort with Type 2 diabetes mellitus (T2DM).

Methods

Baseline data from three trials forming two cohorts, healthy (n = 70) and T2DM (n = 42), were used for combined statistical analyses. Cluster analysis and principal component analysis were used to identify groupings within 10 zinc transporter and MT gene expressions of peripheral blood mononuclear cells, stratified by health status. Multiple regression models were used to explore relationships among zinc transporter/MT groupings, dietary zinc and plasma zinc.

Results

Gene expression of most cellular zinc transporters and MT were lower in the T2DM cohort (P < 0.01). Cluster analysis identified similar groupings of zinc transporters and MT between the cohorts, with the exception of the placement of 2 transporters: ZnT1 and ZIP7. In the T2DM group, ZnT1 was associated with ZnT6 and ZIP3, while ZIP7 was associated with ZnT5, ZnT7, ZIP1 and ZIP10. When grouped by the components identified, zinc transporters and MT were significant determinants of plasma zinc (r2 = 0.48, P = 0.001) in the healthy cohort, but not in T2DM.

Conclusions

The current study suggests altered cellular zinc homeostasis in T2DM; the dissociation of ZnT1 and ZIP7 from their groupings as identified in the AH cohort, highlights potential defects in zinc transport system associated with T2DM.

Support or Funding Information

N/A
Regulation Of Selenogenome, Selenoproteome, And Selenocystenine Biosynthesis And Degradation‐Related Genes By A Novel Organic Se Compound In Avian Tissues
Authors: Sun Lv‐hui , Zhao Ling , Briens Mickael , Xu Shi‐wen , Lei Xin Gen
Published: November 2023

A new organic Se compound, 2‐hydroxy‐4‐methylselenobutanoic acid (SeO), was shown to be more bioavailable than Na2SeO3 or seleno‐yeast (SeY) to chicks, hens, and pigs. This study was to determine regulations of 26 selenoprotein genes, 5 selenoproteins (GPX1, GPX4, SEPP1, SEPW1, and SELU), and 5 selenocystenine biosynthesis and degradation‐related genes (Pstk, SeciSbp2, Sephs1, SepSecS, and Scly) by the three sources of Se in liver, breast, thigh, and/or plasma of chicks. Day‐old male broilers (n = 6 cages/diet, 6 chicks/cage) were fed a Se deficient, corn/soy‐based diet (BD, 0.05 mg Se/kg) or the BD supplemented with Na2SeO3, SeY, or SeO at 0.2 mg Se/kg for 6 weeks. While the three sources of Se enhanced (P < 0.05) mRNA levels of Gpx1, Gpx3, Sepp1, Pstk, and SepSecS in all three tissues over the BD group, only SeO upregulated (P < 0.05) the expression of Sels and Sepx1 in the liver and thigh. Compared with Na2SeO3, SeO caused greater increases (P < 0.05) in expression of Gpx3 mRNA and GPX4, SELU, and SEPP1 proteins in the three tissues and/or plasma. A greater (P < 0.05) expression of GPX4 and SEPP1 protein in the three tissues and/or plasma and Sepp1 and SepSecS mRNA in the two muscles was produced by SeO than SeY. In conclusion, SeO exhibited a unique ability to induce the expression of Sels and Sepx1 mRNA and to enhance the production of GPX4 and SEPP1 proteins in the chick tissues.

Support or Funding Information

Supported in part by NSFC 31501987 and 31320103920, NSTSPC‐2013BAD20B04, and Adisseo.
Dietary excess selenium regulates the transcription of pancreatic selenoproteome in chicken
Authors: Li Jin‐Long , Li Wei , Xiang Li‐Run , Cao Chang‐Yu , Jiang Xiu‐Qing , Zhu Shi‐Yong
Published: November 2023

Background

Recently, numerous studies have found high Selenium (Se) influences the pancreas in mammals. However, little is known about the influence of high Se in the pancreatic tissue in avian.

Objective

We attempted to characterize the relationship between dietary Se, the selenoprotein, and the pancreas in chicken. This project will provide certain help for poultry farming and provide a reference for medicine.

Methods

1‐day‐old chickens received different quantities of dietary Se (0.15 mg, 3.0 mg, or 5.0 mg Se/kg of diet) for 8 weeks. Blood samples were analyzed immediately for glucose content at 2nd, 4th, 6th, and 8th week. The pancreas tissue was collected and examined for the activity of selenoenzymes (GPX, TrxR), and antioxidant related factors (T‐AOC, MDA, GSH) and mRNA expression of the transcriptome of selenoproteins at 4th and 8th week.

Results

In the 5.0 mg Se/kg group, a significant increase (P<0.05) in blood glucose was observed. Dietary excess Se influenced on four antioxidant factors, such as T‐AOC, H2O2, MDA, and CAT. From these data, we observed that excess Se induces decreased antioxidant capacity of pancreatic tissue. At super‐nutritional Se (3.0 mg/kg), most selenoproteins were up‐regulated in chicken pancreatic, but Gpx3, Sepp2, and Selu was only down‐regulated.

Conclusion

Dietary Se level can inluence the pancreatic antioxidant capacity by regulating the activity and transcription of selenoenzymes. These results indicate that excess Se induces pancreatic dysfunction through selenoproteins. Dietary Se status stabilizes pancreatic normal physiology function via regulation of the selenoprotemic transcriptions. We hypothesize that hierarchy of regulated the transcriptions of selenoproteome makes an important roles of renal Se metabolism and transport in avian.

Support or Funding Information

China New Century Excellent Talents in University (No. NECT‐1207‐02), National Natural Science Foundation of China (No. 31572586) and Academic Backbone Project of Northeast Agricultural University (No. 15XG16).
Choline and betaine alter fetal brain expression of genes related to docosahexaenoic acid availability in a mouse model of gestational diabetes
Authors: Joselit Yaelle , Jack‐Roberts Chauntelle , Axen Kathleen , Jiang Xinyin
Published: November 2023

Gestational diabetes mellitus (GDM) is associated with lower cord blood docosahexaenoic acid (DHA) concentrations, which can potentially delay brain development of affected infants. Phosphatidylcholine synthesized via the de novo pathway is enriched with DHA, thereby providing an available source of DHA. Using a mouse model of GDM, we assessed whether supplementation of dams with choline or its oxidative derivative, betaine, modified the expression of genes related to DHA availability in fetal brains. C57BL/6J female mice were fed a high‐fat diet (60% kcal from fat) six weeks before and throughout gestation to induce GDM‐like symptoms. The animals received 25 mM choline chloride, 1% betaine, or control drinking water until embryonic day 17.5. Fetal brain samples were collected for gene expression analysis. Betaine supplementation in the high‐fat fed dams increased the expression of fatty acid binding protein 7 (Fabp7), which transports DHA, and Fabp5, which transports saturated fatty acids in fetal brains. Choline supplementation did not alter the expression of Fabp genes. However, choline upregulated expression of phospholipase A2 (Pla2g6), which releases DHA from the middle position of the glycerol backbone of phospholipids such as phosphatidylcholine, enhancing availability of free DHA in the brain. Expression of arachidonate 15‐lipoxygenase, cyclooxygenase‐2, and brain‐derived neurotrophic factor were not altered by choline or betaine supplementation. Our results support a role for maternal choline or betaine supplementation in modifying DHA availability in fetal brain by altering gene expression in the offspring of GDM dams.

Support or Funding Information

NIH, New York Academy of Sciences, PSC‐CUNY
Effects of Zinc Supplementation on Carnitine and Lactate Concentrations in Cultured Liver Cells
Authors: Barton‐Wechsler Silena , Stachura David L , Ho Emily , Housley Lauren
Published: November 2023

Zinc is an essential cofactor involved in hundreds of biochemical reactions in the human body and serves many critical roles in human health, including the support of energy metabolism. Zinc deficiency is a major public health concern, leading to over 450,000 deaths annually in children under the age of five. Even marginal deficiencies may result in negative health effects if left untreated. Currently, it is challenging to reliably detect and study the health impact of marginal zinc deficiency, because a sensitive, reliable biomarker of zinc status has not yet been characterized. Metabolomics is an emerging tool that has shown promise in biomarker discovery. Analysis from a randomized, placebo‐controlled clinical trial, employing metabolomics analysis, revealed changes in plasma metabolite profiles that identified prospective biomarkers of zinc status. Specifically, our analysis revealed that carnitine (p = 0.04) and lactate (p = 0.02) concentrations were increased in human plasma following zinc supplementation. To further establish if the change in zinc nutriture led to the observed changes in the levels of these energy metabolism‐associated metabolites, we grew human liver cells (HepG2) in media with different levels of zinc to mimic states of zinc deficiency (0.7μM ZnSO4), zinc adequacy (2.9 μM ZnSO4), and zinc supplementation (10.0 μM ZnSO4). One group of cells cultured in zinc‐deficient media was transferred to zinc‐supplemented media to mimic conditions in the previous clinical trial. Following growth in these differential media, carnitine and lactate levels were quantified by colorimetric assays and revealed differential effects of zinc status. This study provides information on the effects of zinc on energy metabolism in humans and informs research aimed at characterizing a sensitive, reliable biomarker for assessing zinc status in humans.

Support or Funding Information

California State University:

Office of Graduate Studies, Research and Conferences Award

Department of Nutrition and Food Science
YAP/TAZ Regulate Hepatic Stellate Cell and Portal Fibroblast Proliferation After Hepatic Ischemia/Reperfusion Injury
Authors: Konishi Takanori
Published: November 2023

Introduction

Hepatic ischemia/reperfusion (I/R) injury is a major complication after liver resection, liver transplantation, and trauma, however, the precise mechanisms of liver recovery after I/R have not been elucidated. The Hippo signaling pathway and their downstream effectors, Yes‐associated protein (YAP) and transcriptional co‐activator with PDZ‐binding motif (TAZ), have been identified as key regulators of regulating cell proliferation and organ size. In the current study, we sought to determine the expression of YAP and TAZ and their contribution to liver recovery after I/R.

Methods

A murine model of partial hepatic ischemia/reperfusion (I/R) was used to study liver recovery after injury. Hepatic stellate cell and portal fibroblast in liver tissue were assessed by immunonhistochemical staining for desmin. Tissue levels of PDGF‐BB and TGF‐b1 were measured by enzyme‐linked immunosorbent assay (ELISA). The expression of YAP/TAZ and their related protein were measured by immunohistochemical staining and Western blot. Proliferation of desmin positive cell was assessed by dual immunofluorescence staining for desmin and proliferationg cell nuclear antigen (PCNA).

Results

Stellate cells and portal fibroblasts, which stained positive for desmin, proliferated around portal veins during early recovery periods. Proliferation of these cells correlated with increased liver expression of PGDF‐BB and TGF‐b1. The expression of phosphorylated LATS1 and phosphorylated MOB1 which sequester YAP/TAZ in the cytoplasm were decreased in liver tissue and nuclear staining of both YAP and TAZ was evident in desmin‐positive cells during liver recovery after I/R. Pharmacological inhibition of YAP and TAZ using verteporfin attenuated desmin positive cell proliferation after I/R.

Conclusion

The data suggest that hepatic stellate cell and/or portal fibroblasts expand during liver recovery after I/R and YAP/TAZ regulate their proliferation.
Thyroid receptor beta agonist reduces tumor burden in an established murine model of human hepatocellular cancer
Authors: Min Qian , Puliga Elisabetta , Columbano Amedeo , Monga Satdarshan(Paul) Singh
Published: November 2023

Triiodothyronine (T3) or selective TRβ agonist GC‐1 induces hepatocyte proliferation at least in part through the activation of Wnt/β‐catenin signaling. This mitogenic effect has formed the basis to support their preclinical use to induce liver regeneration in mice. However, to enable its use in clinical setting, it is highly relevant to directly investigate the effect of compounds like GC‐1 in carcinogenesis especially since Wnt pathway activation due to CTNNB1 mutations is common in hepatocellular cancer (HCC) cases. Our current study was aimed to explore the effect of GC‐1 in preclinical model of HCC driven by CTNNB1 mutations. We co‐expressed hMet and S45Y‐mutant‐β‐catenin in hepatocytes by sleeping beauty transposon/transposase and hydrodynamic tail vein injection, which induces HCC by 4 weeks. At this time mice were randomly stratified into two groups, one that recieved regular diet ad libitum versus second which received GC‐1 diet (5mg/kg) ad libitum. At least 4 mice from each group were euthanized at 10 days and at 21 days after starting the diets. Mice on GC‐1 diet showed a significantly reduced tumor burden as reflected by lower liver weight to body weight ratios at both timepoints. Immunohistochemistry (IHC) for Glutamine synthetase showed fewer and smaller tumor nodules in GC‐1 group. A modest decrease in cyclin‐D1 was visible in GC‐1 group and was shown to occur in tumor nodules where as increased cyclin‐D1 was evident in non tumor areas of the liver. A dramatic decrease in phospho‐Met and phospho‐ERK was clearly evident in GC‐1 treated livers, which coincided with a pronounced decrease in tumor proliferation as detected by both Ki‐67 and Brdu IHC. In conclusion GC‐1 reduces HCC tumor proliferation through blocking hMet signaling while not having any impact on Wnt signaling within tumors. Thus, GC‐1 actually leads to less tumor burden and hence may be safe to use in promoting liver regeneration in chronic liver diseases and transplantation settings.
Substituted Chalcones and Flavones as Human Monoamine Oxidase A (hMAO‐A) Inhibitors with Prostate Cancer Antiproliferative Effects
Authors: Zarmouh Najla O , Gangapuram Madhavi , Eyunni Suresh , Elshami Faisel , Redda Kinfe K , Messeha Samia S , Soliman Karam F
Published: November 2023

Aggressive prostate cancer (PCa) tumors exhibit an increased expression of monoamine oxidase A (MAOA). The use of MAO‐A inhibition as a strategy to treat PCa have not been adequately investigated. In the current investigation, we tested 22 of synthesized chalcone and flavone derivatives for their inhibition of recombinant human (h) MAO‐A activity and their anticancer effect in the PCa LNCaP. Our results showed that five of these derivatives inhibited hMAO‐A with an IC90 < 100 μM, 2‐(3,4,5‐trimethoxystyryl)‐3‐hydroxy‐5,7‐dimethoxy‐4H‐chromen‐4‐one (2a, 6%), 6,8‐dichloro‐3‐hydroxy‐2‐phenyl‐4H‐chromen‐4‐one (2b, 8%), (E)‐3‐(3,4‐dichlorophenyl)‐1‐(2‐hydroxy‐4,6‐dimethoxyphenyl)prop‐2‐en‐1‐one (2c, 9%), (E)‐1‐(3,5‐dichloro‐2‐hydroxyphenyl)‐3‐phenylprop‐2‐en‐1‐one (2d, 3%) and 6,7‐dichloro‐2‐phenyl‐4H‐chromen‐4‐one (2e, 3%), to be comparable to standard MAO‐AIs of pirlindole (4%) and clorgyline (2%). Furthermore, the IC50 of the top compounds was 2e (IC50= 0.03 μM) to be the most potent among all the screened compounds on hMAO‐A. This compound was followed by 2a (IC50= 0.7 μM), 2d (IC50= 1.2 μM), 2c (IC50= 4.9 μM), 2d (IC50= 15.5 μM), respectively. In silico, docking scores were generated for the compounds under study using Open Eye's OEDocking Hybrid method. The highly active compound 2e scored high −7.21 (Chemgauss4 Score) compared to all other compounds under study at the active site of human MAO‐A crystal structure (PDB; 2Z5X). The effects of these five compounds on PCa cells were investigated using the resazurin fluorometric assay. At 20 μM Incubation for 72 h, 2a, 2b, 2c and 2d showed a significant LNCaP cells viability inhibition (P > 0.0001), while 2e was not effective. Compound 2d (IG50= 5.5 μM), 2b (IG50= 8.8 μM), and 2c (IG50= 9.5 μM) were the most potent antiproliferative compounds which were more effective than the standard MAO‐AIs pirlindole and clorgyline (IG50 > 10 μM). The obtained data using 2b, 2c, 2d, and 2e chalcone and flavone derivatives suggest that MAOA has a potential as a therapeutic target in PCa.

Support or Funding Information

Supported by NIH Grants G12 MD007582 and P20 MD 006738
Using Peer‐assisted Learning in a Pharmacology Undergraduate Research Program to Enhance Student Engagement and Performance
Authors: Kemp‐Harper Barbara Kathryn , Drummond Grant Raymond , Davis Elizabeth Anne
Published: November 2023

Introduction

The coursework component of the Pharmacology undergraduate research program at Monash University comprises the critical analysis of a scientific paper under exam conditions. Traditionally, teacher‐led tutorials were utilised to highlight pharmacological techniques, experimental design and data analysis. However, although students were given opportunities to undertake practice exams, overall performance in the assessment task has been relatively poor.

Aims

The current study aimed to revise the learning activities associated with the literature critique exam to encourage student engagement and improve performance.

Methods

In 2013, a tutorial program was implemented which involved peer learning groups such that groups of 4–5 students critiqued components of a scientific paper, presented their findings to the class and lead class discussion. In 2014, to further align the learning activity (group work) with the assessment task (individual work), the exam format was changed to incorporate a peer‐assisted learning component. Students were given the opportunity to discuss the scientific paper with their peers for 30 minutes before being given the exam questions which were then completed under usual exam conditions. The 2014 exam paper was identical to the 2013 paper. In 2015, a new exam paper was used yet the format remained unchanged, with the inclusion of a peer‐assisted learning component. Evaluation of the effectiveness of this change in tutorial and exam format, was via comparison of the exam results and a student feedback survey at the end of the unit.

Results

Students entering the Honours program in 2012–2015 had a similar background knowledge in pharmacology with the average mark in the foundation 3rd year pharmacology unit ranging from 73.2 ± 10% (n=14, 2015) to 78.0 ± 8.1% (n=19, 2012). Following the 2013 revision of the tutorial format from ‘teacher‐led’ to ‘peer‐assisted’, there was no change in the exam marks (2013: 65.7 ± 9.9%; 2012: 63.4 ± 10.2%). However, the majority of students strongly agreed that the peer‐group format of the tutorials enhanced their learning. The introduction of the peer group discussion at the start of the 2014 exam appeared to increase exam performance (74.0 ± 7.3%, n=14), an effect which was sustained in 2015 (72.9 ± 5.3%, n=14).

Conclusion

Our results suggest that the inclusion of peer‐assisted learning in both the learning activities and assessment task may increase the level of engagement of students and improve their critical analysis skills.

Support or Funding Information

Department of Pharmacology, Monash University
Rescue from Synthetic Cannabinoid Intoxication
Authors: Ginsburg Brett C
Published: November 2023

The number of patients seeking emergency treatment for acute synthetic cannabinoid intoxication have increased over the past decade, despite laws prohibiting their legal sale and use. Reported clinical signs of acute synthetic cannabinoid intoxication include tachycardia and hypertension, lethargy, confusion, agitation, hallucinations, and seizures. Clinicians have little guidance on the best course of treatment for such patients. The current standard of care involves administering benzodiazepines to sedate agitated patients and reverse seizures, antidepressants to treat anxiety reported by patients, and antipsychotics to reverse apparent psychotomimetic effects. However, the effectiveness of these treatments remains unclear. Here, in adult male C57 mice, the ability of several clinically available medications to reverse effects including hypothermia, antinociception, and ataxia of the synthetic cannabinoid JWH‐018 are examined. Additionally, effects of JWH‐018 on blood pressure and heart rate are determined, to evaluate potential cardiac effects. JWH‐018 (0.3, 1.0, or 3.0 mg/kg) or vehicle was administered 10‐min prior to test agents which included diazepam (1 & 10 mg/kg), imipramine (10 & 30 mg/kg), fluvoxamine (10 & 30 mg/kg), and haloperidol (0.3 & 3 mg/kg). Effects of these agents are compared to those of rimonabant (1 & 3 mg/kg), the CB1 antagonist. Behavioral effects were assessed immediately before, then 10, 20, 30, 40, and 50‐min after JWH‐018 administration. In other mice, cardiac effects were assessed 25–35 min after JWH‐018 administration. The potential reversal agents are administered after JWH‐018 administration because the clinical situation necessitates administration of reversal agents after cannabinoid exposure. JWH‐018 had no effect on blood pressure and only modestly decreased heart rate. As expected, JWH‐018 had dose‐dependent effects on rectal temperature, antinociception, and ataxia that persisted for at least 50‐min. Within 10–20 min of administration, rimonabant reversed hypothermic and antinociceptive effects and partially reversed ataxic effects of JWH‐018. No other agent tested affected any of the JWH‐018 effects. These results suggest that clinical reports of hypertension and tachycardia are unlikely to be due to direct effects of synthetic cannabinoids, but rather are likely due to the situation arising from intoxication requiring emergency medical care. Further, while treatment with a sedative might help calm an agitated patient, these medications are unlikely to alter the course of synthetic cannabinoid intoxication. Finally, although rimonabant was withdrawn from human use, this decision was based on adverse events during chronic rimonabant treatment for obesity. These results indicate that rimonabant or a similar drug could be useful for acute reversal of synthetic cannabinoid intoxication.

Support or Funding Information

Supported by The Department of Defense (JPC‐5) project FA8650‐15‐C‐6589
Methamphetamine Self‐Administration History May Increase the Reinforcing Effects of Cannabinoid Receptor Agonists in Rhesus Monkeys
Authors: Minkiewicz Molly , John William S , Nader Michael
Published: November 2023

Animal models of drug self‐administration (SA) are a predictive measure of abuse liability. While Δ9‐tetrahydrocannabinol (THC) remains one of the most commonly abused drugs among humans, it has been an on‐going challenge to establish animal models of intravenous SA of THC. In a recent study (John et al., 2017), we found that THC did not function as a reinforcer in rhesus monkeys responding under a fixed‐ratio (FR) 10 schedule of reinforcement, but the cannabinoid receptor (CBR) agonist CP 55,940 was reinforcing in 3 of 8 monkeys; these 3 monkeys all had a previous methamphetamine (MA) SA history. In an effort to increase THC SA in those 8 monkeys, we treated them chronically with THC and found that THC functioned as a reinforcer in 3 monkeys (only 1 of which CP 55,940 had reinforcing effects). The goals of the present study were to (1) evaluate how chronic THC treatment affected CP 55,940 in those 3 monkeys and (2) examine whether exposure to MA would enhance CP 55,940 SA. After a year of abstinence from chronic THC treatment, CP 55,940 (0.001–0.04 μg/kg/injection) no longer functioned as a reinforcer at doses previously identified as reinforcing in those 3 monkeys. To test the hypothesis that the MA history had influenced the reinforcing effects of CP 55,940, these three monkeys and a fourth animal with no previous MA history, were given access to 0.03 mg/kg/injection MA for 8 sessions and then CP 55,940 was re‐evaluated under an FR 10 reinforcement schedule. Following a brief re‐exposure to MA, resulting in intakes ranging from 2.90–6.3 mg/kg, CP 55,940 functioned as a reinforcer in 2 of the 3 monkeys relative to their previous baselines; in the previously MA‐naïve monkey, this history did not increase the reinforcing effects of CP 55,940. CP 55,940 did function as a reinforcer in the monkey with the highest MA intake (6.3 mg/kg), but the sample size is too small to know if there is a direct relation between MA intake and CBR agonist reinforcement. Additional studies are underway to determine if the chronic MA history affected dopamine D3 receptor availability and if this transient increase was necessary for the CBR agonist to have reinforcing effects. Ultimately, our goal is to identify conditions that will permit CBR agonists to function as reinforcers in Old World monkeys.

Support or Funding Information

DA06634
Effects of Brevetoxin and NMDA on Dendritic Arborization in Murine Organotypic Hippocampal Slice Culture
Authors: Gomez Dina , Murray Thomas
Published: November 2023

Background

During neuronal development dendritic arbors become complex as function of both intrinsic genetic factors and neuronal activity. The N‐methyl‐D‐aspartate receptor (NMDAR) is essential for activity‐dependent neurite outgrowth, dendritic arborization and spine formation. Dysfunction in NMDARs is associated with diverse neuropsychiatric disorders such as depression and autism spectrum disorder. High and low NMDAR activity, respectively, leads to excitotoxity and failure of neuronal development. Voltage‐gated sodium channel (VGNaC) activation using gating modifiers such as brevetoxin (PbTx‐2) is one pathway to influence NMDAR function. PbTx‐2 augments NMDAR function in dissociated cerebrocortical neurons. Here we assessed the effect of PbTx‐2 and NMDA on dendritic arbor complexity in organotypic hippocampal slice cultures (OHSC). The activity of the NMDAR is also linked to its phosphorylation status. Phosphorylation of tyrosine residues on the NMDAR increases the open probability and mean open time of the NMDAR leading to increased Ca2+ levels. Calcineurin (CaN) regulates NMDAR function by dephosphorylating the tyrosine residues. In cerebrocortical neurons, FK‐506, an inhibitor of calcineurin, eliminated the reduction in neurite outgrowth observed at high concentrations of PbTx‐2. Here, we determined the influence of FK‐506 alone on dendritic arborization in OHSC.

Hypothesis

1) VGNaC activation or direct activation of NMDARs will promote dendritic arborization in OHSC. 2) FK‐506 alone will not affect dendritic arbor complexity in OHSC.

Methods

The Thy‐1 YFP transgenic mice were used. Hippocampal slices were prepared from postnatal day 5 mouse pups. Slices were treated 18 hours after plating with control, (10–1000 nM) PbTx‐2, (0.01 – 100 μM) NMDA or (0.001–10 μM) FK‐506 for a period of 4 days. A Leica SP8 microscope was used to generate Z‐stack confocal images and Imaris‐XT software was used create 3D‐reconstructed images of the dendritic arbor to run a sholl analysis.

Conclusion

The results demonstrated that both 100 nM PbTx‐2 and 1 μM NMDA treatment produced significant increases in dendritic arbor complexity. The OHSC appeared more sensitive to NMDA than dissociated cerebrocortical neurons inasmuch as 100 μM NMDA was extremely toxic to OHSC neurons. The use of the OHSC confirmed that VGNaC activation augments dendritic arbor complexity and this effect generalizes to hippocampal pyramidal neurons. Our results also indicate that (0.001–10 μM) FK‐506 alone had no effect on dendritic arborization in OHSC. Overall these studies suggest that VGNaC activators may represent a novel pharmacological strategy to regulate NMDAR function and promote neuronal structural plasticity.

Support or Funding Information

NIH‐RO1NS053398
The study of circulating biomarker panels in early diagnosis of Alzheimer's disease
Authors: Zhou Wen‐xia , Jiang Ning , Ruan Can‐jun , Cheng Xiao‐rui , Zhang Yong‐xiang
Published: November 2023

Early, reliable and non‐invasive diagnosis of Alzheimer disease (AD) is a great challenge, which makes the prognosis and therapeutic interventions quite difficult. We aimed to investigate the expressions of let‐7g, miR‐197, miR‐126 and miR‐29a which were screened from 877 microRNAs in 60 serum samples with a microarray platform in prior study in serum of cognitively normal controls (CNC), mild cognitive impairment (MCI), a potential preliminary stage of AD, and AD and identify microRNA panel for predicting and diagnosing AD. Quantitative reverse‐transcriptase polymerase chain reaction assay was applied to evaluate the expressions of let‐7g, miR‐197, miR‐126 and miR‐29a with two independent cohorts including 202 participants CNC, MCI and AD. Logistic regression model based on microRNA panel was constructed using a training cohort (n=150) and then validated using an independent cohort (n=52). Area under the receiver operating characteristic curve (AUC) was used to evaluate diagnostic accuracy. The expressions of the four microRNAs were significantly decreased in MCI and AD versus CNC, and positively correlated with mini mental state examination (MMSE) score. Then, we identified a microRNA panel with the four microRNAs that demonstrated good diagnostic performance for MCI (AUC=0.788 and 0.815 for training and validation data set, respectively) and AD (AUC=0.769 and 0.823 for training and validation data set, respectively). When combined with MMSE score, the diagnostic performance of the microRNA panel was further improved. These results suggested that the serum microRNA panel we found has considerable clinical value in diagnosing AD, and the four microRNAs are potential circulating biomarkers.

Support or Funding Information

The project supported by the National Science and Technology Ministry (2011ZX09102‐101‐07, 2012ZX09301003‐002‐001, 2012BAI29B07)
Rapamycin as a Novel Therapeutic for Alzheimer's Disease: Prevention Assessed Through Neuroimaging
Authors: Wang Amy C , Ma David Wang , Parikh Ishita , Bakshi Vikas , Hoffman Jared , Thalman Scott , Lin Ai‐Ling
Published: November 2023

Alzheimer's disease (AD) is the sixth leading cause of death in the United States. Although AD research is bustling, a concrete and effective treatment has yet to be discovered. This study aimed to assess vascular magnetic resonance imaging (MRI) as an early detection method for AD, and to then test the potential of FDA‐approved drug Rapamycin (Rapa) in treating pre‐symptomatic AD. Rapa is capable of increasing lifespan in mammals and inhibiting unwanted cell mobility, such as cancer cell growth. This study is the first to connect Rapa's capabilities with AD. In order to analyze Rapa's efficacy, neuroimaging and behavioral tests were conducted on mice that carry the apolipoprotein E4 gene (APOE4), the strongest genetic risk factor for AD. Methodology included MRI, magnetic resonance spectroscopy (MRS), Blood‐Brain Barrier (BBB) integrity and neuroinflammation determinations, Radial Arm Water Maze (RAWM), and Novel Object Recognition Test (NOR). These methods provided a comprehensive analysis of the subjects' neurological health and cognitive functions by giving insight into cerebral blood flow, brain metabolites, spatial memory, recognition memory, and neuroinflammation. Vascular MRI effectively pinpointed vascular defects and therefore accurately identified early AD. Data strongly supported the initial hypothesis that with the introduction of Rapa, APOE4 mice will show improved cognition, memory, and neurological health. Cerebral blood flow and crucial brain metabolites were restored in Rapa mice; Rapa mice exhibited cogent memory in comparison to their counterparts in cognitive tests; Rapa reduced neuroinflammation‐‐all with significance. Thus, vascular neuroimaging is a promising early AD detection technique and Rapa holds high potential in preventing AD development.

Support or Funding Information

The work was supported by funding from the National Institute on Aging (NIA) of NIH (K01AG040164 to A‐LL) and NIH CTSA at the University of Kentucky (UL1TR0000117; pilot grant to A‐LL).
Zfra restores memory deficits in Alzheimer's disease triple transgenic mice by blocking tau aggregation and amyloid beta formation
Authors: Lin Yu‐Hao , Shih Yao‐Hsiang , Lee Ming‐Hui , Lin Sing‐Ru , Chang Jean‐Yun , Sze Chun‐I , Kuo Yu‐Min , Chang Nan‐Shan
Published: November 2023

Introduction

Zfra is a naturally occurring 31‐amino‐acid protein. Synthetic peptides Zfra1–31 and Zfra4–10 are known to effectively block the growth of many types of cancer cells.

Methods

10‐month‐old triple‐transgenic (3×Tg) mice for Alzheimer's disease (AD) received synthetic Zfra peptides via tail vein injections, followed by examining restoration of memory deficits.

Results

Zfra significantly down‐regulated tau and amyloid beta (Abeta) aggregates in the brains of 3×Tg mice, and effectively restored their memory capabilities. Zfra inhibited melanoma‐induced neuronal death in the hippocampus and plaque formation in the cortex. Mechanistically, Zfra blocked the aggregation of Abeta42 and many serine‐containing peptides in vitro, and bound cytosolic proteins for accelerating their degradation in ubiquitin/proteasome‐independent manner.

Discussion

Zfra peptides exhibit a strong efficacy in blocking tau aggregation and Abeta formation and restores memory deficits in 3×Tg mice, suggesting its potential for treatment of AD.

Support or Funding Information

Department of Defense USA; Ministry of Science and Technology, Taiwan; National Cheng Kung University Top Notch Project
Co‐ultra PEALut Enhances Neuronal Recovery After a Moderate Traumatic Brain Injury
Authors: Cuzzocrea Salvatore , Paterniti Irene , Campolo Michela , Crupi Rosalia , Filippone Alessia , Cordaro Marika , Bruschetta Giuseppe , Esposito Emanuela
Published: November 2023

Traumatic brain injury (TBI) is a major health problem worldwide. Currently, there is no effective treatment to improve neural structural repair and functional recovery. Recently, has been demonstrated that hippocampal injury‐induced neurogenesis is one mechanism underlying endogenous repair following TBI. The aim of this study was to evaluate the neuro‐regenerative properties co‐ultra PEALut (constituted by the association of palmitoylethanolamide (PEA), with the flavonoid luteolin (Lut)) in a mouse model of TBI until 7 days. TBI was induced in mice by the controlled cortical impact (CCI), one of the most common models of TBI; the mice were orally administered co‐ultra PEALut (1mg/Kg) 1h after trauma and daily for 72h and 7d. A conspicuous neurogenesis was seen in mice 72h and 7d after trauma. Co‐ultra PEALut treatment stimulated neuronal reconstitution process restoring the basal level of new neurons (by BrdU staining) and mature neurons (by doublecortin staining). Moreover this positive effect was associated with an important upregulation of neurotrophic factors (BDNF and NT‐3), ultimately leading to improvement in memory recall on behavioral testing (Morris water maze test).

Co‐ultra PEALut could represent a therapeutic potential of augmenting the endogenous repair response for treating TBI.
Characterization of Rap1A‐dependent activation of PLCɛ
Authors: Sieng Monita , Garland‐Kuntz Elisabeth E. , Lyon Angeline M.
Published: November 2023

Phospholipase C (PLC) enzymes hydrolyze the lipid phosphatidylinositol 4,5‐bisphosphate (PIP2) to produce diacylglycerol (DAG) and inositol 1,4,5‐triphosphate (IP3) which activate protein kinase C and promote intracellular Ca2+ release, respectively. These second messengers are critical for many physiological processes, especially within the cardiovascular system where they regulate cell growth and survival. The PLCɛ subfamily has emerged as a key player in cardiovascular health, due to its involvement in cardiac contractility and regulation of hypertrophic gene expression. PLCɛ is activated downstream of G protein coupled receptors, including the β adrenergic receptors, through direct interactions with small G proteins. The best characterized PLCɛ activator is the small G protein Rap1A. In contrast to canonical PLC activation, Rap1A· GTP binds to PLCɛ and translocates the activation complex to the perinuclear membrane, where phosphatidylinositol‐4‐phosphate (PI4P) is the preferred substrate. Activation by Rap1A is associated with the onset of cardiac hypertrophy. However, while Rap1A is known to interact with the C‐terminal Ras association (RA) domain of PLCɛ, little is known about the molecular basis of activation. We seek to characterize the interactions between Rap1A and PLCɛ using structural and functional studies to map the entire Rap1A binding site on PLCɛ, and determine whether activation results in conformational changes that release autoinhibition and/or increase membrane association. These studies provide the first molecular details of the Rap1A‐dependent activation of PLCɛ.

Support or Funding Information

Funding provided by American Heart Association Grant 16SDG29920017 to Angeline M. Lyon.