bluuebunny/crossref_metadata_2025 huggingface dataset:
Protopanaxatriol‐type of Korean ginseng enhances nitric oxide production in human endothelial cells via genomic and non‐genomic signaling pathway
Authors: Ahn Heeyoon , Hong SoYoung , An Yunhui , Kim JiYeon , Kwon Oran
Published: June 2021

We previously investigated that protopanaxatriol‐type of Korean ginseng (Kg1) stimulated nitric oxide (NO) in human umblilical vein endothelial cells (HUVECs) derived cell line EA.hy926. EA.hy926 was established by fusing primary human umbilical vein cells with a thioguanine‐resistant clone of A549. However, the precise mechanisms in human endothelial cells for Kg1‐mediated nitric oxide production remained unclear. In this study, we hypothesized that Kg1 may induce NO production in human endothelial cells through both genomic and non‐genomic signaling pathways inside the cells. First, to test the genomic regulation of Kg1 on NO production, RT‐PCR analysis was performed for eNOS mRNA expression. It revealed increase of eNOS mRNA in dose‐dependent manners. To examine NO signaling cascade in the cytosol, we used diverse antagonists like L‐NMMA (eNOS), wortmannin (PI3K), RU 486 (GR :glucocorticoid receptor), compound C (AMPK). Also, western blot analysis was performed to test Kg1 regulation on the activations of GR, PI3K(Phosphoinositide 3‐kinase )/Akt or AMPK(AMP‐activated protein kinase). Furthermore, this hypothesis was verified by using the primary HUVECs. Extracellular NO production was measured by Griess reagent and 4,5‐diaminofluorescein (DAF‐2). And image of intracellular NO production was observed by DAF‐2DA fluorescence.
Association between Body Fat and Metabolic Abnormalities in Lebanese Children and Adolescents
Authors: Hwalla Nahla Chawkat , Nasreddine Lara
Published: June 2021

Objective

This study was conducted to investigate the relationship between obesity and various components of the metabolic syndrome (MS) among children and adolescents in Lebanon.

Methods

A total of 260 children and adolescents (161 obese, 50 overweight, 49 normal weight) were recruited from private and public schools in Lebanon. Anthropometric, biochemical and blood pressure measurements were performed.

Results

The number of metabolic abnormalities and their clustering increased with obesity. According to the modified Adult Treatment Panel III definition, the MS was identified in 26.4% and 28.4% of obese children and adolescents, respectively, exceeding rates reported from developed countries such as Italy (13.5%; 16.5%) and Spain (18.0%), while being lower than those reported from the United States (48.8%; 42.0%), the United Arab Emirates (44.0%) and Iran (41.9%).The most common abnormalities among obese children and adolescents were elevated waist circumference (83.9%; 90.5%), high triglycerides (TG) (42.5%; 24.3%) and low HDL (33.3%; 40.5%). The TG/HDL ratio appeared as the strongest predictor of the MS (OR=2.24; OR=4.40).

Conclusion

This study shows that obese children and adolescents in Lebanon may be at increased risk of metabolic abnormalities. The TG/HDL is a useful tool to predict the clustering of cardiometabolic risk factors in the Lebanese pediatric population.
Effect of camel milk on the activities of ATPases in normal and streptozotocin‐diabetic rats
Authors: Alsaif Mohammed A
Published: June 2021

According to the WHO projections, the prevalence of diabetes is likely to increase by 35% by the year 2025. In this study, the streptozotocin (STZ)‐induced diabetic rats, the activities of membrane‐bound adenosine triphosphatases (ATPases) are altered in erythrocytes and in tissues such as liver and kidney. Albino Wistar rats were rendered diabetic by a single intraperitoneal injection of STZ (40 mg/kg body weight). Diabetic rats exhibited significantly (p<0.05) increased levels of plasma glucose and decreased levels of plasma insulin. The activities of total ATPases, (Na++K+)‐ATPase, Ca2+‐ATPase and Mg2+ ATPase were significantly (p<0.05) decreased in diabetic control rats. Control and diabetic rats were treated with camel milk (250 mL/day) for a period of 45 days. A group of diabetic rats were also treated with glibenclamide (600 ìg/kg body weight). After the treatment period, a significant (p<0.05) decrease in the levels of glucose and increase in the levels of plasma insulin and the activities of ATPases in erythrocytes and tissues were observed in diabetic rats treated with camel milk. A similar effect is also observed in the glibenclamide treated rats. But, control rats treated with camel milk did not show any significant (p<0.05) effect in any of the parameters studied. Our study shows that camel milk has the potential to restore the deranged activities of membrane‐bound ATPases in STZ‐diabetic rats.
Associations between blood lipid and inflammatory biomarkers and obesity in university students
Authors: Wang Shu , Reed Debra , Goli Srikanth , Goswami Debalina
Published: June 2021

Obesity is an inflammatory disease. We aimed to explore the relationships between blood lipid and inflammatory biomarkers and obesity in university students. This cross‐sectional study included 110 university students from Texas Tech University. Serum triglyceride and cholesterol concentrations were measured by using enzymatic methods. Plasma concentrations of leptin, adiponectin, C‐reactive protein (CRP), interleukin‐6 and tumor necrosis factor alpha were measured using enzyme‐linked immunosorbent assay. Higher body mass index (BMI) was associated with increased blood concentrations of CRP, leptin, and triglyceride (only in males) and decreased blood adiponectin concentrations in university students. Higher BMI was associated with slightly increased blood cholesterol concentrations in males. Female students had significantly higher blood concentrations of leptin, adiponectin, and CRP than male students. Blood inflammatory biomarkers, especially leptin and CRP, provide more sensitive and accurate assessment than blood cholesterol and triglyceride for obesity in this population. Leptin, adiponectin and CRP are gender‐dependent inflammatory biomarkers.
Ameliorating effects of the Gastrodia elata on scopolamine‐induced amnesia in rats
Authors: Park Eun Ock , Kim Sun‐Young , Back Hyang‐Im , Park Soo‐Hyun , Oh Mi‐Ra , Kim Min‐Gul , So Byung‐Ok , Chae Soo‐Wan , Sui Zhi Yan , Zhao Tong , Huang Guang‐Biao , Chung Young‐Chul
Published: June 2021

We attempted to investigate the effects of GE on scopolamine‐induced amnesia in rats using water maze test. Wistar rats were randomly allocated into 4 groups. Each group received 100, 500, or 1,000 mg/kg of the GE or cellulose daily for two week. After one week, scopolamine (0.5 mg/kg, i.p.) was administered 30 min daily before water maze test. The latency to find the platform was obtained in acquisition trial and working memory task and total time spent in the previous target quadrant was analyzed in probe trial. The control group administered cellulose for 1 week and scopolamine 30 min before water maze test showed significant increases in latency time in both acquisition trial and working memory task and time spent in the previous target quadrant in probe trial. The GE group treated with GE 500 mg/kg for 1 week and scopolamine 30 min before water maze test demonstrated significantly less latency time in acquisition trial compared to the control group. No significant differences were found in latency time in working memory task and total time spent in the previous target quadrant in probe trial. Our results suggest that GE may have ameliorating effects on the scopolamine induced amnesia in rats. Further studies need to be pursued to investigate long‐term effects of GE.
Metabolomic Profiling Reveals Pro‐Inflammatory Lipid Biomarkers Associated with Obesity
Authors: Sampey Brante P , O'Connell Thomas , Vanhoose Amanda M , Kuan Pei Fen , Freemerman Alex J , Newgard Christopher B , Makowski Liza
Published: June 2021

We set out to identify relevant biomarkers associated with obesity by comparing a traditional lard‐based high fat diets (HFD) to those more accurately reflecting poor Western diets of humans. Our previous work demonstrated the robust obesogenic, diabetogenic, and inflammatory outcomes of a nutrient poor, energy dense “Cafeteria diet” (CAF) vs. HFD. The objective of this study was to identify CAF‐related metabolite biomarkers that correlate with physiologic measures of Metabolic Syndrome (MS). Comprehensive metabolomic analysis of several tissues from rats fed CAF, HFD, or controls revealed that multiple saturated fatty acids (SFA), acylcarnitines (AC) and amino acids contributed to the CAF‐effect. Since SFA and AC are known pro‐inflammatory mediators, we performed multiple Spearman pairwise correlations with physiologic and histologic measures. CAF‐associated elevations in markers of adipose inflammation (crown‐like structures) correlated with several ACs including the oxidative intermediate of laurate (C12‐AC). In sum, CAF provides a robust model of human DIO compared to traditional HFD models, resulting in mitochondrial dysfunction and pro‐inflammatory biochemical alterations. Several SFA and C12‐AC emerged as relevant biomarkers of the inflammatory status of adipose tissue and MS, identifying putative targets to limit obesity‐induced inflammation and MS. Funding: K99/R00 AA017376.
Metformin Abolishes Increased Tumor FDG Uptake Associated With a High‐Energy Diet
Authors: Mashhedi Haider , Blouin Marie‐JosÃ© , Zakikhani Mahvash , David StÃ©phanie , Zhao Yunhua , Bazile Miguel , Birman Elena , Algire Carolyn , Aliaga Antonio , Bedell Barry , Pollak Michael
Published: June 2021

Insulin regulates glucose uptake by normal tissues. Although there is evidence that certain cancers are growth‐stimulated by insulin, the possibility that insulin influences tumor glucose uptake as assessed by 18F‐2‐Deoxy‐2‐Fluoro‐D‐Glucose Positron Emission Tomography (FDG‐PET) has not been studied in detail. We present a model of diet‐induced hyperinsulinemia associated with increased insulin receptor activation in neoplastic tissue, and with increased tumor FDG‐PET image intensity. Metformin, which has been associated with reduced cancer burden among diabetics, abolished the increase in insulin levels, tumor insulin receptor activation, and FDG‐PET signal associated with the high‐energy diet, but had no effect on these measurements in mice on a control diet. These findings suggest that for a subset of neoplasms, diet and insulin are variables that affect tumor FDG‐PET results, and have implications for design of clinical trials of metformin as an anti‐neoplastic agent.
Investigation of microRNA‐146a and microRNA‐218 expression in cervical cancer
Authors: Wang Hong‐Wei , Terinate Paul , Gao Youfang , Kalra Krishan L.
Published: June 2021

Background

Cervical cancer is the second most common cancer among women worldwide. However, information about miRNAs in cervical cancer remains limited.

Objective

To examine miR‐146a and miR‐218 expression patterns in cervical cancer, and to determine their value as biomarkers for cervical cancer.

Methods

Cervical cancer tissue microarray (TMA) samples (BioGenex TS‐4215) were used in this study. Probes for miR‐146a and miR‐218 were labeled with 5’‐Fluorescein. Fully automated Xmatrx system (from slides baking, denaturation, hybridization, detection, and final coversliping) was used for the experiments.

Results

Among the 33 cervical cancer TMA samples, upregulated cytoplasmic expression of miR‐146a in cancer cells was observed in 22 tissue samples (66.7%). There were 11 cervical cancer samples that exhibited basal level expression (33.3%). Only 8 out of 33 cervical cancer samples demonstrated elevated levels of miR‐218 (24.2%), 19/33 (58%) showed basal level expression, and there were 6/33 samples (18%) displayed low expression level.

Conclusions

miR‐146a expression is upregulated in cervical cancer. This may be used as a biomarker for the molecular diagnosis of cervical cancer. Future determination of the expression level and clinical prognosis is of importance for developing miR‐146a as a potential novel therapeutic agent against cervical cancer. (Supported by BioGenex Laboratories)
Cell fusion causes DNA double‐stranded breaks and transformation
Authors: Zhou Xiaofeng , Cascalho Marilia , Platt Jeffrey L.
Published: June 2021

Cell fusion promotes nuclear reprogramming, tissue repair and regeneration but may also induce tumors. Whether cell fusion drives tumorigenesis on its own is not known. To address this question, we fused rat intestinal epithelial cells (IEC‐6), which have a normal karyotype, exhibit contact inhibition and anchorage dependant growth, using polyethylene glycol and sorted the hybrids by FACS to establish clonal populations. Clones of single diploid IEC‐6 cells were also established as controls. After ten passages, we determined the fraction of fused clones that grew spheres in soft agar, a measure of cell transformation. More than 25% of the fusion‐derived clones were transformed, compared to 1% of non‐fused IEC‐6 clones. Fusion derived cells exhibited chromosome truncation and translocation, suggesting DNA double‐stranded breaks occurred at the outset or early passage. Consistent with this possibility, fusion‐derived cells studied at early passages had a high rate of DNA double‐stranded breaks, as estimated by γH2AX staining and comet assays. We propose that fusion‐induced DNA double‐strand breaks predispose cells to transformation by increasing the frequency of chromosomal rearrangements. These findings implicate cell fusion, as seen in inflammation, tissue injury and repair, in the genesis of tumors. This work is supported by NIH grant 5R37HL052297.
Quantitative assessment of endogenous retinoids in breast cancer models
Authors: Kane Maureen A , Shao Changxing , Pingitore Attilio , Pierchalski Keely
Published: June 2021

Breast cancer pathogenesis involves disregulation of signaling by retinoic acid (RA), the active metabolite of vitamin A and an essential regulator of cell proliferation, differentiation, and apoptosis. It is not clear if changes to endogenous RA concentrations (via RA biosynthesis defects) or changes to proteins that translate the RA signal drive physiological outcomes. We are interested in understanding if endogenous RA levels are altered in tumors and, if so, when does disregulation of RA concentrations occur during carcinogenesis. From evaluations of gene expression of various components of the retinoid pathway, disregulation of RA signaling is an early event in cancer making the retinoid pathway is an attractive metabolic target for intervention and chemoprevention. Altered expression of genes involved in the trafficking of retinoids is indirectly linked to RA synthesis defects with the implication that RA synthesis defects exacerbate tumor progression. However, there has been no direct quantification of endogenous RA to characterize if RA is altered and, if so, the extent of the alteration. Here we describe quantitative mass spectrometric approaches for quantifying endogenous RA and other retinoids in tissue and present data from models used to study mammary carcinoma.
Increased oxidative stress in patients undergoing coronary artery bypass surgery predicts a higher coronary risk
Authors: Tejerina Teresa , Gonzalez‐Correa Jose‐Antonio , Carnero Manuel , Munoz‐Marin Javier , De la Cruz Jose‐Pedro , Navarro‐Dorado Jorge , Ramajo Marta , Redondo Santiago
Published: June 2021

Introduction

We studied multiple oxidative stress related products in patients who underwent coronary artery bypass grafting (CABG).

Materials

We measured 2 hours before and 24 after CABG the concentration of malondialdehidic acid (MDA), nitrates, and peroxynitrites (prooxidative markers); and reducted glutation and mitochondrial superoxide dismutase (SOD‐Mn) (antioxidative markers).

Results

119 patients were included. Statistically significant differences were detected in the plasmatic MDA concentration before (0.148 mM) and after (0.283 mM) surgery (p<0.001). A trend towards higher concentrations of peroxynitrates (p=0.443) and nitrates (p=0.078) was also noted. A trend towards lower levels of reducted glutation and SOD‐Mn was noted (p=0.94 and p=0.070). Hematite MDA and nitrite levels were predictors of greater Framingham/ATP III score (B=32.146 points/mM/ml (IC 95% 24.132–40.16); B=0.326 points/μM/g (IC 95% 0.049–0.604). GSH levels were inversely related to Framingham/ATP III score (B= −0.175 points/μM/g (IC 95%=−0.344‐ −0.005)).

Conclusions

CABG surgery worsens the oxidative stress profile in patients with coronary disease by increasing oxidation/ antioxidation balance.
Analysis of oligonucleotide DNA binding to hydrotalcites and other nanomaterials
Authors: Sanderson Brian A , Lewis L. Kevin , Beall Gary W.
Published: June 2021

Hydrotalcite (HT) is a naturally occurring double metal hydroxide nanomaterial consisting of magnesium and aluminum ions that exhibits plate‐like morphology. The metal hydroxide layers have a positive charge that is compensated by negatively charged interlayer anions due to isomorphous substitution of Al3+ for Mg2+. These interlayer anions can be exchanged with various external anions, including biomolecules. Biomolecules such as nucleic acids and proteins form strong associations with HT because they can associate with the positively charged layers. The binding of nucleic acids with HT and other nanomaterials is currently being investigated for potential use in gene therapy; however, the binding of specific nucleic acid forms, such as single‐ and double‐stranded DNA, has been little explored. In this project, a series of HTs were prepared at different temperatures and with different anion exchange capacities (milliequivalents per 100 grams). The goal of the project is to investigate the binding of different forms of DNA with each series of HT. Initial experiments have demonstrated that HTs synthesized at higher temperatures associate with both single‐ and double‐stranded DNA more tightly than HTs synthesized at room temperature, likely due to the hydrothermal conditions promoting larger particle sizes.
Effect of enalapril on circulating NO and oxidative stress markers in hypertensive patients
Authors: Silva Pâmela Souza , Fontana Vanessa , Palei Ana Carolina Taveiros , Sertório Jonas Tadeu Cau , Biagi Celso , Tanus‐Santos José Eduardo
Published: June 2021

The aim of this study was to examine the effect of enalapril on plasma levels of markers of NO formation and oxidative stress in mild to moderate hypertensive patients. Eighteen untreated hypertensive patients were treated with enalapril 10 or 20 mg/day for 60 days. Eighteen normotensive healthy controls were followed for the same period. Venous blood samples were collected at baseline and after 30/60 days of treatment with enalapril. Plasma nitrite, NOx (nitrites+nitrates), thiobarbituric acid‐reactive species (TBARS), 8‐isoprostanes and whole blood nitrite concentrations were determined. Treatment with enalapril decreased blood pressure in hypertensive patients. However, we found no significant changes in plasma NOx, nitrite, whole blood nitrite, and in the levels of markers of oxidative stress in both normotensive controls and in hypertensive patients treated with enalapril. Our data show that enalapril 10–20 mg/day does not affect the concentrations of relevant markers of NO formation or markers of oxidative stress in mild to moderately hypertensive subjects, despite satisfactory blood pressure control. Supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).
Effect of high fat diet‐induced type 2 diabetes on liver and serum paraoxonase activities in C57BL/6J mice
Authors: Howell George Eli , Meek Edward , Chambers Janice
Published: June 2021

Diabetes mellitus is the seventh leading cause of death in the U.S. and is a major risk factor for cardiovascular disease. The causative link between type 2 diabetes, which accounts for 90–95% of all cases of diabetes mellitus, and increased prevalence of cardiovascular disease remains elusive. In recent years it has been discovered that the serum level and activity of paraoxonase‐1 (PON1), a cardioprotective enzyme, is decreased in humans with type 2 diabetes. Therefore, the current study explored the effect of high fat diet‐induced type 2 diabetes on PON1 production and activity in C57BL/6J mice. After twelve weeks, animals in the high fat fed group were obese, hyperinsulinemic, hyperglycemic, dyslipidemic, and hyperleptinemic when compared to low fat fed controls. Serum PON activity, as measured by hydrolysis of paraoxon, was significantly elevated in the high fat fed animals while liver PON activity was unchanged. Serum levels of PON1 and apolipoprotein A‐1 protein were comparable between the two groups. However, hepatic PON1 protein levels were decreased in the high fat fed animals. Thus, the current data demonstrate that high fat diet‐induced diabetes in the present murine model increases PON activity in the serum without a corresponding increase in protein production. The present work was supported by a preliminary data grant from the College of Veterinary Medicine Department of Basic Sciences.
Effects of DA11 in db/db Mice and Streptozotocin ‐induced Type 2 Diabetic Rat Model
Authors: Kang Se‐Chan Kang , Yang Yoon‐Jung , Kim Han Kyeol , Kang Nam‐Sung , Sohn Eun‐Hwa , Kim Sun‐Young
Published: June 2021

The Effects of DA11, a medicinal plant mixture developed as a pharmaceutical for the treatment of diabetes mellitus (DM), were investigated in type 2 diabetic db/db mice as well as in streptozotocin (STZ)‐induced diabetic rat models. Various doses of DA11 were administered to the db/db mice and STZ‐induced rats for 5 weeks by PO. The blood glucose levels of the diabetic control animals were increased, whereas the concentrations in DA11‐treated animals remained consistently low throughout the experiment period. Serum insulin levels were not different between control and DA11‐treated animals when measured at the end of the study. DA11 suppressed the levels of total cholesterol and triglyceride after 5 weeks in both the db/db mice and the STZ‐induced diabetic rats. In microscopic observation and immunohistochemistry, DA11 restored the insulin level. These data demonstrate that DA11 can provide beneficial effects in treating type 2 DM by lowering serum glucose levels as well as total cholesterol and triglyceride.
NOVEL AMIDOXIMES THAT INDUCE APOPTOSIS OF COLON CARCINOMA CELLS DECREASE HISTONE ACETYLATION AND INHIBIT p300 IN VITRO
Authors: REDDY SUDHEER REDDY DHANI
Published: June 2021

The eukaryotic nucleus contains chromatin which consists of DNA wrapped 1.6 times around repetitive units of histone octamer called core particles. The folding and unfolding of chromatin can be regulated by the deacetylation and acetylation of lysine residues on the histones, respectively. Histone acetyltransferases and histone deacetylases are two groups of enzymes that modulate the degree of acetylation of histones, therefore regulating the levels of gene expression. Our project tested the growth inhibition effect of nine novel amidoximes on six malignant cell lines. Four of the compounds showed a specific and significant inhibition of proliferation in several malignant cell lines. DNA fragmentation assay and cleavage of caspase 3 demonstrated that these compounds also induced apoptosis in colon cancer cells. These compounds inhibited the acetylation of specific lysines on core histones in colon cancer cells. In vitro acetylation assays revealed that one of the amidoximes, JJMB9, is an inhibitor of the histone acetyltransferase p300. These results show for the first time to our knowledge, amidoximes that demonstrate HAT inhibition activity and anti‐proliferative effect.
Synthesis of Some New 3‐(Heteroaryl‐iminoethyl)‐Benzopyran‐2‐ones and their Antimicrobial Activity
Authors: Hoti Ramiz , Mulliqi‐Osmani Gjyle , Vehaoi Idriz , Thaçi Veprim , Gashi Fatbardh , Ismaili Hamit
Published: June 2021

Novel 3‐(heteroaryl‐iminoethyl)‐benzopyran‐2‐ones are synthesized by catalytic condensation of 3‐acetyl‐benzopyran‐2‐one and corresponding heteroarylamines. Condensation of 3‐acetyl‐benzopyran‐2‐one 1 and 2‐aminothiazoles 2(a,b) gave corresponding 3‐[(thiazol‐2‐ylimino)‐ethyl]‐benzopyran‐2‐ones 3(a,b). By catalytic condensation of 3‐acetyl‐benzopyran‐2‐one 1 and 3‐amino‐5‐methylthio‐1H‐1,2,4‐triazol 2c, 3‐[1‐(5‐methylthio‐1,2,4‐triazol‐3‐ylimino)‐ethyl]‐benzopyran‐2‐one 3c was synthesized, whereas condensation of 1 and 3‐amino‐5,6‐dimethyl‐1,2,4‐triazine 2d gave 3‐[1‐(5,6‐dimethyil‐triazine‐3‐ylimino)‐ethyl]‐benzopyran‐2‐one 3d. The antimicrobial activity of products 3(a–d) were investigated and results were submitted for their activities against E. coli, S. aureus and Candida albicans. Compound 3c showed light bactericide activity against S. aureus. Emphatic activity against E. coli exhibited compounds 3a and 3b, whereas 3d and 3c exhibited the strongest activity against Candida albicans. In general there was a direct positive correlation between increasing concentration of the compound and antimicrobial activity.
Analgesic and Anti‐Inflammatory Effect of Pyridazinones/Aminoantipyrine Hybrids
Authors: mavaneh khatereh fattahpour , Inceler Nazan , Baytas Sultan Nacak , Gokce Mehtap , Uludag M. Orhan , Abacioglu Nurettin
Published: June 2021

In this study we have investigated the analgesic and anti‐inflammatory effect of synthesized 6‐subsititue‐3(2H)‐pyridazinone/aminoantipyrine hybrids. Twelve pyridazinones/aminoantipyrine hybrids were synthesized and evaluated for their in vivo analgesic and anti‐inflammatory activity using p‐benzoquinone‐induced writhing test and carrageenan‐induced paw edema model, respectively. 30 min after the subcutaneous administration of a pyridazinones/aminoantipyrine hybrid (100mg/kg body weight), the mice (n=8) were injected with 0.1mL/10g body weight of 2.5% (w/v) p‐benzoquinone solution intraperitoneally and suspension of carrageenan (0.5mg/25μL) into sub‐plantar tissue of the right hind paw. The total number of the abdominal contractions (writhing movements) was counted for the following 15 min, starting 3 min after the p‐benzoquinone injection. Paw edema was measured in 90, 180, 270 and 360 min after carrageenan injections. Among the synthesized compounds, especially, SNB 144, NI 29, NI 31, NI 33 showed potent anti‐inflammatory activity and SNB 156, SNB 140, SNB 146 showed potent analgesic activity as compared to indomethacine (10mg/kg), aspirin (100mg/kg) respectively. Therefore, we can conclude that the preparation of suitable pyridazinones/aminoantipyrine hybrids of contemporary NSAIDs may represent a potentially useful method for developing compounds with potent analgesic and anti‐inflammatory activity.
EFFECT OF CLOZAPINE ON THE ENTERIC SYSTEM: IMPLICATIONS FOR ANTIPSYCHOTIC‐INDUCED WEIGHT GAIN
Authors: Palavicino‐Maggio Caroline B , Tobon Krishna , Douard Veronique , Ferraris Ronaldo P. , Kuzhikandathil Eldo V.
Published: June 2021

It is well established that the use of atypical antipsychotics results in weight gain and metabolic diseases such as obesity, type‐2 diabetes and metabolic syndrome. However, the underlying mechanisms for weight gain are not clear. Recent studies have shown that various dopamine receptor subtypes are expressed in the enteric system; however, the physiological role as well the effect of antipsychotics on the enteric dopamine receptors is not well understood. We hypothesized that the dopamine receptors might be involved in the modulation of nutrient transport in the intestine and alteration of this modulation by chronic antipsychotic treatment might contribute towards weight gain. In particular, excessive and chronic intake of fructose is known to adversely affect long‐term health and lead to obesity, type‐2 diabetes and metabolic syndrome. The intestinal fructose transporter GLUT5 is considered as a biomarker of excessive fructose consumption. In this project, we test the hypothesis that antipsychotics acting at D2‐like dopamine receptors modulate the expression or function of intestinal GLUT5 transporters, resulting in weight gain. To test this hypothesis, we compared the ability of clozapine (CLZ) to induce weight gain in wild type C57BL/6 mice and GLUT5‐null mice. Preliminary results showed that CLZ dose‐dependently induced weight gain in the wild type C57BL/6 mice but not in the GLUT5‐null mice. In addition, there was a dose‐dependent increase in fructose uptake in the C57BL/6 wild type mice treated with CLZ. The expression of GLUT5 mRNA was not altered by CLZ treatment suggesting that the increase in fructose uptake induced by CLZ might be mediated by an increase in GLUT5 activity. The results suggest that modulation of intestinal nutrient transport by antipsychotics might contribute to weight gain.
Sex‐specific gene responses to estrogen and androgen in human osteoclasts
Authors: Stern Paula H , Wang Jun
Published: June 2021

Estrogen and androgen are both critical for maintenance of bone, but the target cells, mechanisms and responses could be sex‐specific. To compare sex‐specific actions of estrogen and androgen on osteoclasts (OC), mononuclear precursor cells from adult Caucasian males (n=3) and females (n=3) were differentiated into OC and then treated for 24 h with 10nM 17β‐estradiol (E2) or testosterone (T), or ethanol (control). Gene expression was studied with a custom designed qPCR array containing 94 target genes related to bone and hormone action. Genes changed > 2 fold with p<0.05 were considered significantly modified. In untreated OC, 4 genes showed significant gender differences. E2 significantly affected 12 genes in OC from females and 6 genes in OC from males. 15 of the 18 E2‐responsive genes differed in cells from the two sexes. 2 of the genes affected by E2 in both sexes were regulated oppositely in the two sexes. T significantly affected 6 genes in OC from females and 2 genes in OC from males; all except one gene were different in the two sexes. The findings indicate that 1) OC from both sexes are affected by both E2 and T; 2) within the genes examined, more were affected in OC from females than males; 3) effects of both E2 and T differ in the two sexes; 4) E2 and T largely affect different genes, suggesting that conversion of T to E2 has a limited role in the gene responses.

Support: Northwestern Univ. Institute for Womens Health Research.
A gamma scintigraphic clinical study of the absorption of Insulin co‐formulated with Eligen® absorption enhancer 4‐CNAB
Authors: Castelli M. Cristina , Connor Alyson , Wilding Ian , Riley M. Gary
Published: June 2021

Orally non‐bioavailable peptides eg. calcitonin, GLP‐1 and insulin are rapidly absorbed when formulated with Eligen carriers. A single center non‐randomized non‐blinded 2‐way crossover study in 6 healthy subjects assessed the bioavailability of insulin deposited directly into the jejunum or dosed orally. Blood glucose and PK of C‐peptide and the Eligen carrier 4‐CNAB were also measured. Regimen A: 150U insulin+80 mg 4‐CNAB as an immediate release (IR) granulate was deposited into the jejunum with an Enterion capsule. Regimen B: The same dose of IR granulate insulin and 4‐CNAB was dosed orally as an 111In radiolabeled tablet. Tablet transit /disintegration and location of Enterion capsule activation were measured scintigraphically. Mean Cmax and AUC0–2 for total insulin were 118 μU/mL and 84 μU*hr/mL respectively following gastric delivery and 38 μU/mL and 49 μU*hr/mL with jejunal delivery. 4‐CNAB and C‐peptide PK were similar with both regimens. Decreased glucose (−35%) accompanied only gastric delivery. No severe or serious adverse events occurred. This study suggests that gastric uptake of insulin occurs when insulin is formulated with 4‐CNAB.
Common alleles in human N‐acetyltransferase 1 (NAT1 *10 and *11) increase enzyme activity via distinct mechanisms and associate with sulfamethoxazole induced hypersensitivity
Authors: Wang Danxin
Published: June 2021

N‐acetyltransferase 1 (NAT1) metabolizes drugs and environmental carcinogens, affecting drug response and cancer susceptibility. Two common NAT1 alleles, *10 and *11, have been reported to alter enzyme activity compared to wild‐type NAT1 (*4), but their effects and underlying mechanisms remain uncertain. We measured allelic NAT1 mRNA expression and translation, considering multiple transcription start sites, alternative splicing, and three 3′‐polyadenylation sites in human livers and B lymphocytes. *11 was found to favor usage of the second 3'polyadenylation site, increasing formation of NAT1 mRNA with the 3'UTR of intermediate length (major isoform). This major 3'UTR isoform supported more efficient protein translation than the short 3'UTR isoform, resulting in increased *11 NAT1 enzyme activity. Similarly, NAT1 *10 also increased protein translation efficiency, but without affecting NAT1 mRNA levels or 3′‐UTR polyadenylation site usage. B‐lymphocytes with *11/*4 and *10/*10 genotypes displayed higher NAT1 enzyme activity than the reference genotype *4/*4. Moreover, carriers of *10/*10 and *11/*4 (‘fast NAT1 acetylators') were less likely to develop hypersensitivity to NAT1/NAT2 substrate drug sulfamethoxazole, but this was observed only in subjects with slow NAT2 acetylator genotype. In conclusion, NAT1 *10 and *11 significantly increase NAT1 enzyme activity, enabling the classification of carriers into normal and rapid acetylators. Supported by NIH grant R21 AI074399 and U01 GM092655.
Interaction of 5‐HT3R ligands with acetylcholine binding protein by surface plasmon resonance
Authors: Rideout Shane Alexander , Harms‐Smyth Abe E , Ataian Yeganeh , Schulte Marvin K
Published: June 2021

The field of Ligand Gated Ion Channels (LGIC) has benefited greatly over the last several years due to the discovery of the Acetylcholine Binding Protein (AChBP) from snail Lymnaea stagnalis and Aplysia californica. Studies have utilized docking simulations, crystal structure and radio ligand binding data of the AChBP to better understand the function of ligand binding. The AChBP has also been used to characterize other cys‐loop super family LIGC including the serotonin type 3 receptor (5‐HT3R). Although homology models for the AChBP and 5‐HT3R exist there is a lack of information describing how 5‐HT ligands interact with the AChBP.

Understanding how 5‐HTR ligands interact with the AChBP binding pocket is crucial knowledge for comparing the functional relationship between AChBP and 5‐HT3R ligand binding site. The goal of this work was to obtain binding data for 5‐HT3R ligands using radio‐ligand binding experiments and Surface Plasmon Resonance (SPR) as well as confirm validity of 5‐HT3R antagonist binding models based on AChBP. We evaluated the binding of 5‐HT3R agonists, partial agonists and antagonists to the AChBP. Results showed binding affinities for 5‐HT3R agonist were 500–30,000 times lower on the AChBP compared to 5‐HT3R. Binding affinities for 5‐HT3R antagonists were 2–30 lower on the AChBP compared to the 5‐HT3R (>KD). The interaction of antagonists to AChBP is similar to those present in 5‐HT3R while those residues interacting with agonists show less homology. Our docking studies supported these conclusions.
CB1 Cannabinoid Receptor Location and CRIP1a
Authors: O'Neal Jeremy , Blume Lawrence C. , Bass Caroline E. , Howlett Allyn C.
Published: June 2021

CB1 cannabinoid receptor (CB1R) is a Gi/o linked GPCR that influences psychological processes such as mood and memory. Cannabinoid Receptor Interacting Protein (CRIP1a) has been shown to interact with the C‐terminal tail of the CB1R, leading to the hypothesis that CRIP1a could influence receptor localization and trafficking. The goal of this study was to identify the role of CRIP1a in CB1R localization using N18TG2 wild type (WT) and CRIP1a over‐expressing neuroblastoma cell clones as a model system. Cells were transfected with green fluorescent protein‐tagged CB1 receptors (GFP‐CB1R). GFP‐CB1R and CRIP1a were imaged using confocal microscopy, and the localization was identified using nuclear (HCS nuclear mask) and early endosome (Cell‐Lights) molecular probes. N18TG2 WT cells displayed CRIP1a uniformly in the cell cytosol. At basal conditions, the GFP‐CB1Rs in WT cells are primarily expressed on the plasma membrane surface. On cell Western studies using an N‐terminally‐directed CB1 antibody, revealed that stable over‐expression of CRIP1a reduced the density of CB1R's on the plasma membrane, when compared to WT cells. During agonist (WIN55212‐2)‐challenge, internalization of CB1 receptors occurs in WT cells, but not in CRIP1a‐overexpressing clones. Upon exposure to the CB1 antagonist rimonabant, the density of cell‐surface CB1 receptors increased in both WT as well as CRIP1a‐overexpressing cells. These studies suggest the CRIP1a influences CB1R trafficking and localization during basal and agonist conditions, and provides a mechanisms for regulating CB1 receptor availability at the plasma membrane.

Supported by: GM‐GM064249, RO1‐DA03690, R21‐DA025321
Oxidative stress induces increases in the gene expression and protein levels of SIRT2 in primary murine astrocyte cultures and PC12 cells
Authors: Nie Hui , Hong Yunyi , Ma Yingxin , Shao Jiaxiang , Chen Heyu , Jiang Jingwen , Xia Weiliang , Ying Weihai
Published: June 2021

Oxidative stress is a major pathological factor in multiple major neurological diseases. Certain Sirtuin family proteins, such as sirtuin 1 (SIRT1), also play important roles in aging and cell survival. In this study we investigated the effect of oxidative stress on the gene expression and protein levels of sirtuin 2 (SIRT2) in primary murine astrocyte cultures and PC12 cells. Our Western blot studies, using the samples collected at 24 hrs after H2O2 treatment, showed that treatment of primary astrocyte cultures with 0.1, 0.3 and 0.5 mM H2O2 increased SIRT2 levels by 100% ‐‐‐ 300% in astrocytes; and treatment of PC12 cells with 0.1, 0.3 and 0.5 mM H2O2 increased SIRT2 levels by 30% ‐‐‐ 80% in the cells. Our real‐time PCR study, using the samples collected at 12 hrs after H2O2 treatment, has also suggested that 0.1 and 0.5 mM H2O2 increased the SIRT2 mRNA levels by approximately 100% in astrocytes. Our immunocytochemistry study has further provided preliminary results suggesting that H2O2 can induce an increase in SIRT2 levels in PC12 cells. Collectively, our study has suggested that oxidative stress can significantly increase the gene expression and protein levels in primary astrocyte cultures and PC12 cells, which may be involved in oxidative stress‐induced cell injury (supported by a National Key Basic Research ‘973 Program Grant’ #2010CB834306, a Pujiang Scholar Program Award 09PJ1405900, and a Key Research Grant of Shanghai Municipal Educational Committee #09ZZ21).
Procyanidin B3, an inhibitor of histone acetyltransferase, enhances the action of antagonist for prostate cancer cells via inhibition of p300‐dependent acetylation of androgen receptor
Authors: Choi Kyung‐Chul , Yoon Ho‐Geun
Published: June 2021

Increasing evidence suggests that AR (androgen receptor) acetylation is critical for prostate cancer cell growth. In the present study, we identified Pro‐B3 (procyanidin B3) as a specific HAT (histone acetyltransferase) inhibitor. Pro‐B3 selectively inhibited the activity of HATs, but not other epigenetic enzymes. Pro‐B3 substantially inhibited the p300‐mediated AR acetylation, both in vitro and in vivo. Pro‐B3 inhibited both p300‐dependent and agonist‐induced AR transcription. We demonstrate that the p300‐mediated AR acetylation is critical for the hormone responsiveness of AR. Interestingly, B3 treatment efficiently enhanced the antagonist activity of flutamide through suppression of p300 HAT activity, demonstrating that relative p300 activity is critical for the antagonist action. Finally, Pro‐B3 treatment inhibited acetylation‐dependent prostate cell proliferation and expression of cell‐cycle control genes, subsequently increasing cell death, indicating the functional importance of AR acetylation for prostate cancer cell growth.
CB1 Cannabinoid receptor Regulates Ethanol‐mediated inhibition of Neurogenesis
Authors: Mukhopadhyay Somnath , Devkota Shailendra , jha Shalini
Published: June 2021

CB1 cannabinoid receptor (CB1R) activation stimulates neurogenesis in developing rat brain. Chronic ethanol treatment has been shown to inhibit developmental and adult brain neurogenesis. The objective of the present study is to determine if CB1R regulates ethanol‐mediated inhibition of developmental neurogenesis in zebrafish. We have used green fluorescence protein (GFP)‐expressing transgenic zebrafish (ISLET‐1) embryos to study neurogenesis using confocal microscopy. We have shown that zCB1R mRNA transcript is expressed in the different regions of zebrafish brain at 48 and 72 hpf (hour post‐fertilization). We also showed that CB1R agonist CP55940 (1–10 μM) treatment stimulated zCB1R expression and motor neurogenesis in zebrafish brain. We further showed knocking down CB1R using morpholino or antagonist‐mediated blockade of CB1R attenuated CB1R‐mediated increase in neurogenesis. Further we have found that chronic ethanol (0.5%–1.5%)‐treatment during embryonic development (6hpf–36 hpf) dose‐dependently reduced zCB1R expression and motor neurogenesis. Co‐treatment with CB1R agonist CP5540 or FAAH‐inhibitor URB597 attenuated ethanol‐mediated inhibition of developmental neurogenesis. Together, results from this study showed that zCB1R plays a vital role in the regulation developmental neurogenesis as well as ethanol‐mediated deleterious effects on neurogenesis.
Heroin self‐administered female rats display impaired memory and increased heroin‐seeking behavior
Authors: Ye Guiping , Wang Jianhua , Zhou Xiaoli , Zhou Xin , Lai Miaojun , Zhu Huaqiang , Fu Dan , Zhou Wenhua
Published: June 2021

Heroin dependence is frequently accompanied with cognitive dysfunction and psychiatric symptoms, but the impact of heroin self‐administration on these remains elucidated. Adult female rats with or without ovariectomy (VOX) were trained under the fixed‐ratio 1 (FR1) schedule for self‐administration of heroin (0.05 mg/infusion, i.v., 4 h/d×14 d). Nose pokers were extinguished prior to reinstatement testing, which consisted of presentation of drug‐paired cues or a priming injection of heroin (0.25 mg/kg, s.c.). After 2 wk of heroin withdrawal, rats were examined in a variety of behavioral tests, including object recognition and fear condition. All the rats produced escalation of daily heroin intake. During reinstatement testing following exposure to conditionedcues and drug‐priming injection with cues, VOX females produced a robust reinstatement behavior. Compared to normal females, VOX rats displayed more susceptibility to memory deficits after heroin self‐administration. These results suggest that ovarian hormones deficit may enhance drug‐primed reinstatement and facilitate cognitive deficits and other psychiatric disorders, such as anxiety and depression, after chronic heroin exposure in female rats.
Behavioral phenotype of phosphodiesterase 4A (PDE4A) knockout mice suggests a role in memory and anxiety
Authors: Hansen Rolf , Wu Qiang , Wang Chuang , Huang Ying , Hu Wei , Jiang Youming , Meng Zhenzhi , Conti Marco , O'Donnell James , Zhang Hanting
Published: June 2021

Phosphodiesterases (PDEs) comprise a group of 11 families of enzymes that play a significant role in the regulation of intracellular signaling. Of particular interest to the CNS is PDE4 (comprised of four subtypes A, B, C and D) due to its high expression in the brain. While the examination of PDE4B and PDE4D knockout mice has begun to elucidate their roles in CNS function, the role of PDE4As remains largely unknown. The purpose of this study was to analyze the behavioral phenotype of PDE4A knockout mice, in particular as it relates to memory, anxiety, and depression. Using a battery of behavioral paradigms, including passive avoidance, water‐maze, forced‐swim, tail‐suspension, elevated plus‐maze, light dark transition, and holeboard tests, we found that PDE4A knockout mice displayed enhanced memory in the passive avoidance test, but not the water‐maze test; the mice also exhibited anxiogenic‐like behavior, but no significant changes in the forced‐swim or tail‐suspension tests. These results suggest that PDE4A may play a significant role in mediating memory and anxiety, but not depression related behavior.
A long‐term efficacy of allogeneic bone marrow mesenchymal stem cells in myocardial repair
Authors: Huang Xi‐Ping , Sun Zhao , Miyagi Yasuo , Zhang Li , Weisel Richard , Li Ren‐Ke
Published: June 2021

Allogeneic mesenchymal stem cells (AMSCs) are currently undergoing clinical trials for myocardial repair. However, their long‐term therapeutic efficacy has not been established. We conducted a six month comparative study on allogeneic and syngeneic MSCs using a MI rat model. We found AMSCs were immunoprivileged soon after implantation and produced a robust cardiac functional improvement similar to syngeneic MSCs in the first three months after treatment. After two weeks implantation, the differentiated AMSCs acquired immunogenic phenotype associated with increased cell surface expression of MHC class I and II molecules, rendering them immune rejected and diminishing their ability to prevent heart failure after 6 months of treatment. These results suggest that AMSCs have short‐term benefits for heart repair, and reconcile inconsistent findings from earlier studies by identifying the unknown biphasic immune response to AMSCs. Differentiation induced immune rejection supports the need for future studies to modify the late immunogenic phenotype of MSCs to prevent rejection and achieve long term benefits of AMSCs for cardiac repair. Supported by HSFO and CIHR.
Establishing a Hierarchy of cis‐Linked Chromatin Structures that May Contribute to Epigenetic Risk
Authors: Mussar Kristofer Joseph , Meagher Richard
Published: June 2021

While a clear cause‐and‐effect relationship is well established between genotype and inheritance of risk factors for human disease, the link between cis‐linked chromatin structures (epitypes) and the risk of acquiring disease is much less clear. This is in part due to the fact that neither the rules nor the rates governing the evolution of epitypes are well understood. There are two ways to establish that particular epitypes have the potential to contribute to the inherited risk of disease. Traditionally, the transgenerational inheritance of risk factors and epitypes in related individuals are measured directly. In addition, risk factors that are transgenerationally inherited may be identified by analyzing the conservation of epitypes after gene duplication. Using these data we've established a hierarchy of epigenetic controls based on their potential to contribute to risk. DNA sequence changes would reside at the pinnacle of the risk pyramid evolving in parallel with epigenetic risk. Below would be other transgenerationally inherited properties like nucleosome position and cytosine methylation. Finally, histone modifications appear to be the least likely to be transgenerationally inherited, and hence, therefore are at the bottom of the pyramid. One outcome of this research is establishing a clear distinction between inherited epigenetic risk and somatically inherited epigenetic factors contributing to organ and cell development. This study is funded by NIH research grant GM‐36397 and training grant GM‐07103.