bluuebunny/crossref_metadata_2025 huggingface dataset:
Activation of Brain Wnt signaling in vivo: Effect on LTP and Neurogenesis
Authors: Inestrosa Nibaldo C
Published: June 2021

The roles of the Wnt signaling pathway in several developmental processes, including synaptic differentiation, are well characterized. Wnts have a crucial role in synaptic physiology, as they modulate the synaptic vesicle cycle, the trafficking of neurotransmitter receptors and the interaction of these receptors with scaffold proteins in postsynaptic regions. Recent studies indicate that Wnts regulate synaptic plasticity and neurogenesis. We report here that the in vivo brain application of both agonists and modulators of Wnt receptors, results in improvement of several behavioral assays, concomitant with an increase in several pre and post‐synaptic proteins as well as in synaptic plasticity measured as long ‐ term potentiation (LTP) in hippocampal slices. These results were obtained with both canonical and non‐canonical Wnt modulators, including WASP‐1 and Foxy‐5. More recent studies, includes the effect of natural products as well as Wnts ligands on adult mammalian neurogenesis. In particular, we have examined the role of hypoxia on the activation of canonical Wnt signaling. Current studies are aim to ask whether non‐canonical Wnt signaling affects neurogenesis, since recent work indicated that Wnt‐5a modulates recycling of functional GABAA receptors in hippocampal neurons.
A Pilot Study: Soy protein may help decrease energy intake when consumed prior to meal
Authors: Cope Mark B , Jenkins Alexandra , Mukherjea Ratna , Krul Elaine , Hughes Glenna , Pawlik Kate , Campbell Janice , Wolever Thomas
Published: June 2021

Protein & fiber reduce subjective appetite ratings acutely, leading to subsequent decreased energy intake in studies with large amounts of protein & fiber. The objective of this study was to measure satiety & energy intake following consumption of snack bars containing modest quantities of soy protein, SP, (20 g), & soy fiber, SF, (8 g) and in combination. In a randomized, double blind crossover design, 40 healthy subjects were assigned to 4 treatments with 4 types of snack bars: Control (C), SP, SF, & SP + SF. Appetite effects & bar palatability (Visual Analog Scale) & post‐meal ad libitum energy intake were measured. Subjects were asked to consume 1 type of bar within 15 mins at each visit (1‐week washout between visits). Overall appetite scores were not significantly different; however, SP tended to decrease post‐meal ad libitum energy intake vs C (−43 kcal). C bars took a significantly longer time to consume than SP or SF bars (10.3 min vs. 7.7 & 8.4 min, respectively). A post hoc sensory analysis of the bars, showed significantly different chewing attributes. The lower energy intake in the SP group is consistent with previous studies & may have long term benefits.
Activation of angiotensin type 2 receptor rescues high‐fat dietinduced adipocyte size increase in young male mice
Authors: Nag Sourashish , Khan Mohammed Azhar , Hussain Tahir
Published: June 2021

Recent studies demonstrated that genetic deletion of angiotensin type 2 receptor (AT2R) decreases adipocyte size and controls obesity, a major risk factor for metabolic syndromes. Current study was designed to test whether pharmacological manipulation of AT2R regulates adipocyte size. Male C57BL/6 mice (4‐weeks old) were pretreated with AT2R antagonist (PD‐123319, 30 μg/kg/min, osmotic pump) or AT2‐R agonist (C21, 0.3 mg/kg, daily i.p.) for 4 days. Thereafter the animals were placed on normal chow diet (ND) or high‐fat diet (HFD) with concurrent drugs treatment for next 10 days. The PD treatment increased the ratio of total abdominal fat weight to body weight in both ND and HFD groups compared with the respective controls. Similarly, the PD treatment caused increase in epididymal fat weight in both ND and HFD. Epididymal adipocyte size was also increased in HFD group treated with PD. On the other hand, C21 treatment rescued HFD‐induced increase in epididymal fat weight and adipocyte size. The HFD‐induced increase in plasma free fatty acid level was prevented by C21 treatment. In sum, pharmacological blocking of AT2R increases and activation of AT2R decreases adipocyte size. This is the first study which demonstrates that adipocyte size increase is rescued by the pharmacological activation of AT2R, which may serve as therapeutic target for controlling obesity and associated disorders.

Grant Funding Source : NIH RO1 DK‐61578
Abdominal obesity and inflammation in tri‐ethnic population with and without type 2 diabetes in South Florida
Authors: Gundupalli Deva , Zarini Gustavo G. , Vaccaro Joan A. , Huffman Fatma G.
Published: June 2021

The aim of the present study was to assess the association of abdominal obesity as measured by waist circumference (WC) with inflammation as measured by high sensitivity C‐reactive protein (hs‐CRP) in a population with and without type 2 diabetes (T2D). Data from a cross sectional study of three ethnic groups (Cubans=304, Haitians=237, African Americans=188) recruited from Miami‐Dade and Broward counties in South Florida was used for the analysis. Hs‐CRP values > 10mg/L (n=136) were excluded. Exploratory data analysis, correlations and linear regressions were run. Hs‐CRP was log transformed to achieve normality. Age, gender, ethnicity, smoking, NSAID use, diabetes status and insulin use were used as control variables in the analysis. WC explained 11.1 % (β=0.36, P<0.001) of the variability in hs‐CRP. The model was significant [F (9, 719)=17.40 (P<0.001)]. Males had lower hs‐ CRP (β=−0.13, P<0.001). Cubans had higher hs‐CRP compared to Haitians (β=0.14, P<0.001). The results indicate that abdominal obesity is a risk for inflammation. Further studies are needed to assess the effect of nutritional intervention on abdominal obesity and inflammation. This study has been funded by NIH/NIDDK grant.
Brown adipocyte commitment of primary human adipose stem cells in vitro
Authors: Chung Soonkyu , Okla Meshail , Ha Jung-Heun , Lee Mee Ae
Published: June 2021

The bone morphogenic protein 7 (BMP7) has shown to promote brown adipogenesis from the adipose stem cells suggesting “Browning of WAT” would be a novel strategy to reduce obesity. However, extrapolating these findings to human is limited due to the developmental differences of adipose stem cells between rodent and human, and a lack of human brown adipocyte model system. The aim of this study was to establish the in vitro model of human brown adipocyte using BMP7. Adenoviral delivery of BMP7 to human adipose stem cells (hASC) induced a dose‐dependent increase of UCP1, a signature protein of brown adipocytes. Brown adipocyte differentiation was also confirmed by a significant increase of brown adipogenic markers, and a decrease of white adipogenic marker leptin. However, PRDM16 which is known to regulate brown adipogenesis was not associated UCP1 expression. Intriguingly, transient COX2 expression preceded the brown adipogenesis and the chemical inhibition of COX2 dampened UCP1 expression. In summary, we 1) established an in vitro cell model of brown adipocyte from the primary human adipose stem cells via adenoviral delivery of BMP7, and 2)demonstrated that COX2 activity is required for brown adipogenesis. These data suggest that BMP7 signaling may activate COX2‐dependent prostaglandin production in human adipose stem cells, which subsequently plays a key regulatory role for the development of brown adipocytes
Black pepper consumption does not influence 24‐hour energy expenditure or respiratory quotient in overweight postmenopausal women
Authors: O'Connor Annalouise , Corbin Karen , Nieman David , Swick Andrew
Published: June 2021

As traditional recommendations have not successfully dampened rising obesity rates, there has been an increased interest in the role of thermogenic ingredients in weight management. It has been suggested that black pepper (BP) may impact energy expenditure (EE), however despite promising in vitro and rodent work, no study has examined the thermogenic capacity of BP in humans. Therefore, our aim was to determine the impact of acute BP consumption on 24‐hour EE, respiratory quotient (RQ), and biochemical markers associated with metabolic rate and satiety in humans. In a randomized cross‐over study, post‐menopausal overweight or obese women (n=17) spent two 24‐hour periods within the metabolic chamber at the UNC Nutrition Research Institute. During each study day, subjects remained sedentary and consumed meals tailored to their energy requirements, supplemented with or without 1.5g of BP (0.5mg/meal). Blood was sampled 30 minutes following lunch for assessment of glucose, insulin, gut peptides (including GLP‐1, PYY, ghrelin), leptin, adrenaline and noradrenaline. Total 24‐hour EE and RQ did not differ significantly between the BP and control study day. There were no significant effects of BP on the biochemical markers assessed. In conclusion, acute consumption of BP did not significantly affect EE or RQ, however, it is possible that longer term treatment and/or combining BP with other spices could increase EE.
Neuroprotective Effects of Papaya Epicarp Extract against Aluminum‐Induced Oxidative Stress in Human Neuronal (SHSY5Y) Cells
Authors: Waly Mostafa , Guizani Nejib , Ali Amanat , Rahman Mohammad Shafiur , Deth Richard
Published: June 2021

Background

Aluminum (AL) as an environmental toxin induces oxidative stress and impairs folate‐dependent methionine synthase (MS) activity in human neuronal cells, SH‐SY5Y cells.

Objective

This study aimed to assess the in‐vitro antioxidant properties of the papaya epicarp extract (PEE) against AL‐induced insults in SH‐SY5Y cells.

Results

The analyzed data revealed that PEE protects against AL‐induced cells death, and in a dose‐dependent manner it restores the inhibited folate‐dependent MS activity and antioxidant enzymes (Catalase, Glutathione Peroxidase and Superoxide Dismutase). In addition PEE augments the AL‐induced impairment of cellular glutathione, thioredoxin and total antioxidant capacity.

Conclusion

This study shows that PEE supplementation has a neuroprotective role in ameliorating the changes in redox status of neural cells exposed to AL, a well known environmental toxin that has been involved in the pathogenesis of neurodegenerative diseases and neurodevelopmental disorders.
Mechanism of D‐psicose absorption is through glucose transporter or not: in vivo and in vitro studies
Authors: Moon MinSun , Jang Han I , Kim You Jin , Kim Ji Yeon , Kwon Oran
Published: June 2021

D‐psicose is a “rare sugar” present in small quantities in nature. Commercially, D‐psicose obtains from the hydrolysis of sucrose or isomerization of D‐glucose. In previous study, mechanisms of absorption on D‐psicose were not investigated yet. So, we evaluated some of the dominant intestinal transporters in vivo and in vitro models. We examined inhibition of 2‐deoxyglucose(DG) or fructose uptake in caco‐2 and AtT20ins. Caco‐2 cells showed inhibition of 2‐DG and fructose uptake. Although the effect was small, AtT20ins was similar to caco‐2 cells. In vivo study, the animals were fed three experimental diets, control(C), 58% high fructose(HF), and 58% high fructose with 5% d‐psicose(HFP) diets for 15 weeks. The body weight gain, fat pad, leptin, and MCP‐1 were significantly lower in HFP than in HF. Despite of no significant changes, insulin and HOMA‐IR were decreased in HFP than in other groups. These novel findings add evidence that the unabsorbed d‐psicose competes with fructose and/or glucose for intestinal glucose transporters which is consistent with in vitro and in vivo studies. *This study was supported by the 2nd stage of Brain Korea 21 Project in 2011 and grant from CJ Co., Ltd.
Single Nucleotide Polymorphisms in the β‐carotene 15,15′‐monooxygenase coding gene and Health Outcomes in Korean Adults
Authors: Kim You Jin , Moon Min Sun , Yang Yoon Jung , Kim Ji Yeon , Kwon Oran
Published: June 2021

In Republic of Korea, major source of dietary vitamin A intake is plant (vegetable, seaweed and fruits). Such as β‐Carotene, provitamin A carotenoid is the primary plant‐based source of vitamin A. β‐Carotene 15,15′‐monooxygenase (BCMO1) is the key enzyme responsible for conversion of dietary provitamin A carotenoids to active form of vitamin A, retinals, in the small intestine. In in vivo and human studies, it was shown that single nucleotide polymorphisms (SNP) in the BCMO1 gene would be affect efficacy of conversion and production of vitamin A. Two nonsynomous SNP (Arg267Ser A>T: rs12934922; Als379Val C>T: rs7501331) was identified the common variants in the coding regions of this gene. In Caucasians, the combined two variant alleles had a frequency of 36% in the HapMap database, whereas 9% in the Han Chinese and 2% in Japanese population. However, BCMO1 SNP frequencies and β‐carotene metabolism had not been investigated in Korean population. So this study was conducted to determine of allele frequency in the Korean population and, further, elucidate the association between allele variants and epidemiological data (anthropometric indexes and blood profile). To investigate, statistical analysis was performed using data from 2 epidemiologic studies: 1) the Ansung‐Ansan cohort study (n=2,000), 2) a cohort of elderly subjects (n=340). * This study was supported by the 2nd stage of Brain Korea 21 Project in 2011.
Pomegranate vinegar feeding enhances fatty acid oxidation: In vitro and in vivo
Authors: PARK YEOJIN , Ok Elly , Lee Hyo Jung , Kim Ji Yeon , Kim Mi Kyung , Kwon Oran
Published: June 2021

Vinegar is not only commonly consumed as a seasoning and a food preservative but also used as a traditionally folk remedy from the ancient. Recent studies show an acetic acid, major component of vinegar, has beneficial effect on anti‐obesity by increasing fatty acid oxidation in liver. In this study, we screened which vinegar beverages contained acetic acid and flavonoids showed synergistic effect on stimulation of genes related fatty acid oxidation using HepG2. We selected pomegranate vinegar beverage as anti‐obesity beverage and then treated to high fat diet rats for 11weeks. After treatment, in liver, muscle, and adipose tissue, we examined several genes related on fatty acid oxidation and synthesis such as PPAR‐a, CPT‐1, ACO, UCP‐2, SREBP using pomegranate vinegar and acetic acid in animal models using RT‐PCR. And also, with western blotting in same animal models, we examined P‐AMPK/ AMPK activation in liver.

*This study was supported by the 2nd stage of Brain Korea 21 Project in 2011
Influence of Fructose Consumption on Inflammatory Gene Expression in Liver and Brain in a Pig Model of Juvenile Obesity
Authors: Jang Saebyeol , Molokin Aleksey , Laskman Sukla , Panickar Kiran , Bruna Gonzalo , Urban Joseph F , Solano-Aguilar Gloria
Published: June 2021

Fructose intake has been associated with the prevalence of overweight and obese humans. Therefore, we studied the metabolic effects of added fructose consumption in young pigs as a model of juvenile obesity. Conventional crossbred pigs were randomized and fed 1) a basal calorie diet alone or 2) with added fructose (20% of total calories); or 3) a high calorie diet with added fat alone or 4) with added fat and fructose for 12 wk. Two‐way ANOVA revealed a significant main effect of fat and a fructose by fat interaction on body weight gain showing that fructose consumption increased body weight gain only when added to a basal calorie diet. Gene expression was increased for hepatic inflammatory genes TNF‐α, IL‐1β, COX‐2 and MCP‐1, and brain hippocampal IL‐6 by high fat consumption whereas gene expression for hippocampal brain derived neurotrophic factor (BDNF) was diminished. There was no significant effect of fructose on gene expression. Interestingly, correlation z test revealed a significant relationship between concentration of serum non‐esterified fatty acid (NEFA) and gene expression of hippocampal IL‐6 and BDNF suggesting that NEFA might be predictive of changes in gene expression in the brain. Overall, this study showed that increased fructose in the diet contributed to body weight gain in a lean condition, but did not negatively affect hepatic and hippocampal‐derived inflammatory gene expression.
Stunting in the first year of life is associated with unfavorable lipid profile in early childhood
Authors: Hoffman Daniel J. , Vitolo Marcia Regina , Campagnolo Paula Dal Bo
Published: June 2021

The objective of this study was to determine the association between stunting and adverse lipid profiles in preschool children. Data were collected during a randomized trial conducted in Sao Leopoldo, Brazil. At 12–16 months, anthropometric data were obtained and cut‐offs for severe stunting were height‐age‐Z score < −2 and mild stunting was < −1.65. Total Cholesterol (TC), Low‐density lipoprotein (LDL), high‐density lipoprotein (HDL) and triglyceride (TG) concentrations were assessed using standard methods at ages 3–4 years. Children who were severely stunted had a TC 14.3 mg/dL higher and non‐HDL cholesterol 13.8 mg/dL higher compared to normal height children, adjusting for gender, waist circumference, maternal education, breastfeeding, and maternal BMI. Children who experienced mild stunting had a TC 12.7 mg/dL higher, LDL‐c 10.2 mg/dL higher, and non‐HDL cholesterol 12.9 higher compared to normal height children. Based on these data, nutritional challenges early in life appear to be associated with an unhealthy lipid profile as early as the preschool years. Funding was provided by the Brazilian National Funding for Research and the Brazilian Ministry of Education.
Age‐dependent expression of retinoic acid‐related orphan receptor‐alpha (Rora) in the developing mouse cortex and hippocampus
Authors: Taniguchi Saori , Armoskus Chris , Tsai Houng-Wei
Published: June 2021

We investigate if retinoic acid‐related orphan receptor‐α (Rora) is sexually dimorphic in developing mouse cortex and hippocampus. Recent study shows autistic patients express lower Rora mRNA in their cortex and cerebellum than controls do. Rora may cause sexual bias in autism susceptibility because it regulates expression of aromatase that converts testosterone to estrogen. Reduction of Rora may decrease aromatase levels to increase testosterone levels in autistic subjects (Sarachana et al., 2011). We predict male mice express lower Rora in the cortex and hippocampus as compared to females on the day of birth (PN0), 7 days after birth (PN7), 14 days (PN14), and 21 days after birth (PN21). We extracted RNA from cortexes and hippocampi of male and female C57BL/6J mice at PN0, PN7, PN14, and PN21 (n=8 per group). We measured mRNA levels of Rora and β‐actin (Actb) by RT‐qPCR, normalized Ct values of Rora and Actb, and analyzed two‐way ANOVA (sexes vs. ages) with Bonferroni's test for post‐hoc comparisons. Rora levels in the cortex and hippocampus were significantly higher in PN14 and PN21 mice than in PN0 and PN7 mice (p<0.05), but there was no significant difference between the sexes (p=0.183). Thus, Rora levels in the mouse cortex and hippocampus in early development is age‐dependent, but not sexually dimorphic. This research is supported by CSUPERB Faculty‐Student Collaborative Research Seed Grant and CSULB startup fund.
EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR 2 CORRELATES WITH METASTATIC POTENTIAL IN PROSTATE TUMOR CELLS
Authors: Brown Graham William , Shimizu Akio , Coma Silvia , Adam Roalyn , Bielenberg Diane
Published: June 2021

Studies in recent years have suggested that VEGFR2 could be expressed in non‐endothelial cell populations. This study explores the expression and function of VEGFR2 on non‐endothelial prostate tumor cells of differing metastatic potential. PC3M and PC3M‐LN4 human prostate carcinoma cell lines, low and high metastatic respectively, were analyzed using RT‐PCR, western blot, ELISA, and in vitro growth assays. Additionally, PC3M and PC3M‐LN4 cells were injected orthotopically into the prostate of male nude mice for in vivo tumor growth analysis. PC3M‐LN4 cells showed expression of VEGFR2 mRNA in RT‐PCR and VEGFR2 protein in western blot analysis whereas PC3M cells did not. ELISAs of PC3M and PC3M‐LN4 cells showed that both cells lines produce high levels of VEGF. Culturing cells in serum free media containing only VEGF‐A resulted in the proliferation and growth of PC3M‐LN4 cells but not PC3M. Tumors resected from nude mice injected with PC3M tumor cells were small and non‐metastatic whereas PC3M‐LN4 tumors were large, highly vascularized, and with metastases to the para‐aortic lymph nodes. These results pose an autocrine growth function for VEGFR2 in highly metastatic prostate tumor cells as well as a correlation between VEGFR2 expression and metastatic potential in these cells. Clinically, these data suggest that VEGFR2 inhibition may target both the tumor and stromal compartments to inhibit disease progression.
A novel role for thiol isomerase ERp72 in platelet activation, thrombosis and hemostasis
Authors: Wang Lu , Zhou Junsong , Wu Yi
Published: June 2021

A recent study has demonstrated that ERp72 is expressed in human platelets. We here investigated its function in platelet activation and thrombosis. We generated a rabbit polyclonal antibody to ERp72 (RP‐anti‐ERp72) using a recombinant full‐length human ERp72 as an immunogen. By Western blot, this antibody specifically recognized human ERp72 antigen, but not other thiol isomerase such as PDI or ERp57. In a concentration‐dependent manner, RP‐anti‐ERp72 markedly inhibited platelet aggregation induced by collagen (2 μg/ml) and thrombin (0.5 U/ml), as well as PAC‐1 binding (an integrin αIIbβ3 activation marker). It also inhibited P‐selectin expression on platelets activated by thrombin and convulxin, a collagen receptor GPVI agonist. As found by flow cytometry, both convulxin and thrombin increased surface exposure of ERp72 on platelets. RP‐anti‐ERp72 recognized ERp72 in mouse platelets in Western blot. When infused into mice, RP‐anti‐ERp72 (200 μg/mouse) increased the occlusion time by >2 fold (12.28 ± 1.96 vs. 4.17 ± 1.48 min, compared with control IgG, P<0.05) in a FeCl3‐induced carotid artery injury model, and prolonged the tail bleeding time (17.16 ± 1.21 vs. 10.32 ± 2.35 min, P<0.05). Our results demonstrate for the first time that a specific member of thiol isomerase family, ERp72, plays an important role in thrombosis and hemostasis, and its effect is at least via the regulation of platelet activation.
Evaluation of the Antithrombotic and Hemorrhagic Effects of a Novel Ultra Low Molecular Weight Heparin (ULMWH)
Authors: Gray Angel , Hoppensteadt Debra , Jeske Walter , Fareed Jawed
Published: June 2021

Semuloparin (S) (Sanofi‐Aventis, Paris, France) is an ULMWH exhibiting high anti‐Xa (aXa) activity due to preservation of antithrombin‐binding sites. Antithrombotic (jugular vein clamping model) and hemorrhagic (tail transection model) effects of S, the LWMH enoxaparin (E) and heparin (H) were compared in Sprague‐Dawley rats treated subcutaneously (SC) with doses of 1.0 and 2.5 mg/kg (n=5). Blood samples collected prior to euthanasia were analyzed using aPTT, aXa and Heptest (HT) assays. 1.0 mg/kg SC H significantly prolonged bleeding time compared to control (C). All agents had equivalent antithrombotic effects. S and E significantly increased HT prolongation and aXa activity compared to C and H (HT: p<0.001 A vs H; p<0.05 E vs H; Xa: p<0.001 A vs H; p<0.01 E vs H). At 2.5 mg/kg SC, only S did not significantly prolong bleeding time compared to C. Only H significantly increased aPTT compared to C. Prolongation of HT and aXa activity was observed with all agents. S exhibited a more potent aXa activity than H (p<0.001 S vs H). All agents demonstrated a dose‐dependent antithrombotic effect. Bleeding time was lower with S at both dosages, suggesting a better safety profile. AXa activity with S was closely related to its antithrombotic activity. These studies suggest that S represents a new alternative for the prevention of thrombotic disorders with an improved beneﬁt/risk proﬁle compared with standard H and E.
Exchange transfusion therapy and its effects on real‐time microcirculation in pediatric sickle cell anemia patients
Authors: To Wilson James , Miller Joshua W. , Miguelino Maricel G. , Li Jiajing , Lin Xin , Chen Peter , Green Ralph , Cheung Anthony T.W.
Published: June 2021

The goal of exchange transfusion therapy is to ameliorate vasoocclusion and improve tissue perfusion by removing sickled red blood cells and introducing normal red blood cells. Using computer‐assisted intravital microscopy, pre‐transfusion and post‐transfusion microvascular characteristics were analyzed and compared. In this study, the bulbar conjunctiva exhibited a “blanched” avascular appearance in all 6 pediatric sickle cell anemia patients before transfusion, indicative of tissue hypoperfusion and ischemia. Immediately after transfusion, substantial improvement in vascularization and tissue perfusion resulted. In addition, a decrease in red cell velocity was observed. These observations provide evidence that exchange transfusion therapy is beneficial in ameliorating vasoocclusion and improving tissue perfusion. However, with the paradoxical post‐transfusion decrease in red cell velocity presumably due to induced hyperviscosity from the large transfusion volume, blood flow is still impaired. This decreased velocity may thwart efforts to improve oxygen delivery through transfusion and may, to some extent, promote vasoocclusion instead.
Clozapine induces mitochondrial alterations and the secretion of proinflammatory cytokines in insulin‐sensitive cell types
Authors: Contreras-Shannon Veronica , Heart Dylan L. , Maffi Shivani , Catano Gabriel , Walss-Bass Consuelo
Published: June 2021

Clozapine is an effective treatment for schizophrenia but causes side effects like weight gain and diabetes. Mitochondria dysfunction, oxidative stress and inflammation are mechanisms proposed for clozapine side effects as well as the development of obesity and insulin resistance. Prior investigations have focused on the brain and neurons. In this study, the effect of clozapine on insulin responsive cell types and cells associated with obesity was examined. Cultured myoblasts, adipocytes, and hepatocytes were treated with 0, 25, 50 and 75 μM clozapine. After 24 hours, a mitochondrial selective probe was used to assess morphology and membrane potential (ΔΨm). ATP lysates were also prepared and culture media collected for cytokine analyses by multiplex array. Clozapine caused mitochondria fission in all cell types. In addition, it altered mitochondria ΔΨm (p<0.05), volume (p<0.05), and ATP levels (p<0.05) relative to controls. Clozapine also significantly induced the production of the proinflammatory cytokines MCP‐1, IL‐6, and GM‐CSF. In conclusion, clozapine damages mitochondria and promotes inflammation in insulin responsive cells. Understanding how clozapine alters metabolism may provide insight on the origin of metabolic disease in the general population. Supported by Biaggini Research Fellowship, St. Mary's Univ. to VC‐S & Friends for Psychiatric Research Grant, Dept. of Psychiatry to CW‐B.
FoxO3a transcription factor differentially modulates the metastatic potential of ER+ and ER− breast tumors
Authors: Morelli Catia , Maris Pamela , Anselmo Wanda , Cesario Maria Grazia , Lanzino Marilena , Sisci Diego , Andò Sebastiano
Published: June 2021

FoxO3a (F3a) transcription factor has been reported to suppress estrogen(E2)‐dependent breast cancer cell growth and tumorigenesis through its functional interaction with the estrogen receptor α (ER). However, the role of this interplay in tumor metastasis is poorly defined and is the aim of the present study. To this purpose, F3a and its active form AAA, were over‐expressed both in ER+ and in ER− breast cancer cell lines. Our results show decreased migration and invasion of F3a/ER+ cells compared to control, paralleled by a strong upregulation of Caveolin‐1 (Cav‐1), the main constituent of caveolae that seems to function as a negative regulator of cell transformation and invasion in breast cancer, and a significant reduction of MMP‐13 mRNA. F3a mediated effects were more evident in AAA expressing cells and were emphasized by E2. On the other hand, in ER− cells, F3a and AAA increased cell migration and invasion with a concomitant induction of MMP‐13 transcripts, while no difference was observed in Cav‐1 levels. Since both Cav‐1 and MMP‐13 promoters bear Forkhead canonical motifs next to ER binding sequences, our hypothesis is that F3a exerts a protective role in ER+ breast cancers through F3a/ER mediated regulation of Cav‐1 and MMP‐13, while it could represent a bad prognostic factor in ER− breast tumors. Financial support has been provided by AIRC (Associazione Italiana Ricerca Cancro).
Programmed cell death by S. pneumoniae D39 and its PspA mutant in microglial and neuronal cells
Authors: KIM JI YUN , Rhee Dong-kwon , Pyo Suhkneung
Published: June 2021

Pneumococcal infection in the central nervous system (CNS) such as brain can trigger apoptosis of neurons and other cells in CNS. Pyroptosis is a caspase‐1‐dependent inflammatory form of programmed cell death. PspA of S. pneumoniae is one of surface virulence proteins which prevents binding of C3 onto pneumococcal surface. The role of PspA in apotoisis or pyroptosis remains unexploited. In the present study, we investigated the apoptotic and pyroptotic effect of S. pneumoniae D39 and its PspA mutant(PspA−) in neuroblastoma cells and microglial cells. MTT assay and western blotting showed that D39 and PspA−, regardless of p53 gene status, induced apoptosis of neuroblastoma cells in a time‐dependent and dose‐dependent manner. Moreover, D39 and PspA− did not cause an activation of caspase‐3 and caspase‐8. Our data also demonstrated that apoptosis inducing factor(AIF) was translocated from mitochondria to the nucleus in neuroblastoma cells treated with D39 and PspA−, respectively. In addition, western blot analysis revealed that treatment of PspA− led to caspase‐1 activation in microglial cells. Thus, these results suggest that Streptococcus pneumoniae D39, and PspA− trigger a p53‐independent and caspase‐independent apoptosis in human neuroblastoma cells and PspA− might have a role in pyroptosis of microglail cells.
Prostate specific antigen: Enhancing the anti‐microbial response of the prostate during urinary tract infection
Authors: Townes Claire Louise , Ali Ased , Lanz Marcelo , Gross Naomi , Robson Craig , Pickard Robert , Hall Judith
Published: June 2021

The inability to successfully clear an initial episode of mild cystitis may result in a greater susceptibility to recurrent urinary tract infection (rUTI). A chart review carried out at our institution showed a significantly increased risk of recurrent urinary tract infection (rUTI) in men whose prostate specific antigen (PSA) levels during an initial UTI episode were low (<4ng/ml). PSA is known to function as a serine protease and several proteins belonging to the WFDC (WAP Four Disulphide Core) family and present in semen are known to contain antimicrobial domains released by serine protease cleavage. We suggest that PSA may function to digest these proteins during UTI and therefore enhance the prostates antimicrobial response.

The expression of genes encoding WFDC2 and SLPI were identified in prostate tissues by RT‐PCR. WFDC2 and SLPI expression (p<0.001) and PSA synthesis (p<0.01) were significantly increased in LnCAP and Du145 prostate cell lines challenged with Escherichia coli, as determined by realtime PCR and ELISA. The presence of PSA significantly reduced the ability of bacteria to invade prostate cells. Incubation of PSA with recombinant WFDC2 and SLPI for 24 hours resulted in total proteolysis of the full‐length protein and significantly increased antimicrobial activity. These results suggest PSA may have an important role in protecting against rUTI.
HMGB1 guides cytokines in early carbon nanoparticle lung exposure
Authors: Herndon Betty , Das Monisha , Nalvarte Elisabet , Molteni Agostino , Quinn Tim , Fibuch Eugene
Published: June 2021

An initial effect of carbon nanoparticle (CN) lung exposure is the cellular release of damage‐associated molecular pattern recognition molecules (DAMPs). One of the earliest is the high mobility group box 1 (HMGB1) protein, which, extracellularly, exerts its actions as a ligand for pattern recognition receptor (PRR) target cell activation. This work studies HMGB1's role in damage recognition and activation of cellular processes. CN (0.025 mg, endotoxin free in 0.050 mL Survanta) was intratracheally instilled into rat airways (n=45). At necropsy 0.5hr to 4 wks later, bronchoalveolar lavage fluid (BAL), lungs and sera were measured (ELISA, cytology) for HMGB1 and 3 cytokines. BALs were also used to activate PRRs in RAW Blue cells, stably transfected lung macrophages expressing the SEAP gene inducible by NF‐kB and AP‐1. HMGB1‐containing BALs induced significant expression of SEAP in the RAW blue cells, highest at 24 hr, which was compared to purified HMGB1 dose‐dependent activation. Histopathology described Inflammation of lung parenchyma and translocation of CN from airways to the mesothelial lining. IL‐10 and IL‐6 in lung related in timing to HMGB1‐receptor complexes (p=0.04). The results show that sterile CN exposure releases lung cell HMGB1, and BAL from this exposure activates raw blue cells at times when HMGB1 is elevated, with related receptors and cytokines.

Sponsor: Saint Luke's Hospital Foundation
A Novel ER Export Motif Modulates the ER‐to‐Cell Surface Traffic of α2B‐Adrenergic Receptor
Authors: Dong Chunmin , Nichols Charles D , Wu Guangyu
Published: June 2021

The molecular mechanisms underlying export of the G protein‐coupled receptor (GPCR) superfamily from the endoplasmic reticulum (ER) remain largely unknown. Here we have demonstrated that a triple Arg (3R) motif in the third intracellular loop functions as a novel ER export signal for α2B‐adrenergic receptor (α2B‐AR). The 3R motif mediates α2B‐AR interaction with Sec24C/D which are components of ER‐derived COPII transprot vesilces, and modulates ER exit, cell surface transport, and function of α2B‐AR. Furthermore, export function of the 3R motif is independent of its position within α2B‐AR and can be conferred to CD8 glycoprotein. These data provide the first evidence implicating that export of GPCRs is controlled by code‐directed interactions with selective components of the COPII transport machinery.
Elucidating the Molecular Mechanisms of the Inhibitory Effects of Viscolin on Neointimal Hyperplasia
Authors: Hsieh Yi-Ni , Wu Chieh-Hsi , Wu Tian-Shung
Published: June 2021

The aim of this study was to elucidate the active compounds of natural products with anti‐inflammatory and anti‐proliferation effects to prevent neointimal formation in rats. Viscolin and MA‐1 have been identified to most significantly suppress the proliferation and migration of vascular smooth muscle cells (A7r5). The anti‐proliferative effects of viscolin or MA‐1 on A7r5 were quantified after 24 h of cell incubation by MTT assay. The anti‐migratory activities of viscolin or MA‐1 on A7r5 were evaluated by a wound migration assay after 24 h and 48h of drug incubation. The anti‐inflammatory effects of viscolin were determined by measuring the pro‐inflammatory mediators in lipopolysaccharide (LPS)‐stimulated murine macrophage RAW264.7 cells. NO concentrations were measured in the culture supernatants via the Griess reaction and ELISA assays. We assessed the effects of viscolin on LPS‐induced iNOS and COX‐2 protein level expression in RAW 264.7 cells. We also determined the effects of viscolin on MMP‐9 and MMP‐2 protein level expression in A7r5 cells by Western blotting. The inhibitory effects of viscolin on neointimal formation in balloon injured rat model were also studied. Viscolin or vehicle were delivered to rats by intraperitoneal injection for two weeks after balloon injury. Morphological analysis using H & E staining at day 14 indicated that viscolin prevented intimal hyperplasia in rat thoracic aorta as compared to vehicle. Our current data suggest that viscolin could be a useful drug to prevent restenosis after balloon injury of the arteries.
TLR4 Overexpression After Orthotopic Liver Autotransplantation Contributes To Acute Lung Injury In Rats
Authors: Chi Xinjin , Ge Mian , Zhang Ailan , Luo Gangjian , Xia Zhengyuan , Hei Ziqing
Published: June 2021

Acute lung injury(ALI) is a common and severe complication of orthotopic liver transplantation(OLT), while the underlying mechanism of post‐OLT ALI is not very clear. Toll‐like receptor (TLR) 4 is an important TLR family member that plays a key role in the innate immune system. TLR4 is increased after OLT, accompanied with increased inflammatory response and ALI (Chin Med J, 2009,122:895–899). We hypothesized that TLR4‐mediated inflammation response may play a ctitical role in ALI during OLT. Healthy SD rats (n=8/group) were randomly divided into sham operated control group (C) and OLT groups with different durations (4, 8 and 16 hours) of reperfusion in the presence or absence of TLR4 siRNA interference performed 48 hours before OLT. Lung tissue TLR4 gene and protein expressions were significantly increased 8 hours after OLT and onwards, which was coincident with increased tissue levels of tumor necrosis factor (TNF) α, interleukin (IL)6 and IL‐1β, and evident lung pathological damage manifested as increased lung injury scores and increased lung water content and functional impairment manifested as reduced PaO2 (all P<0.05 vs. OLT vs. C). TLR4 dominant negative siRNA prevented all the above changes after OLT. It is concluded that TLR4 overexpression, potentially by stimulting proinflammatory cytokine over production, plays a critical role in the developent of ALI after OLT.

The study was supported by National Natural Science Foundation of China grant (No. 30801080 and 30972858).
Thyroid hormone (T3) regulates lipoprotein metabolism via induction of Angiopoeitin‐like 4 (ANGPTL4) in human hepatoma cell line
Authors: Talasila Phani K. , Sadana Prabodh
Published: June 2021

Angiopoietin‐like protein 4 (ANGPTL4) is known to be expressed in several tissues, including the metabolically active liver as well as adipose tissue. There is strong evidence of ANGPTL4 playing a role in lipid metabolism through its inhibition of the activity of Lipoprotein Lipase (LPL). Triiodothyronine (T3) is a hormone known to play a critical role in metabolic homeostasis especially in the lipid metabolism. Thyroid hormone mimetics are under clinical development to treat hypercholesterolemia because of their ability to reduce LDL and total cholesterol levels in the plasma. We hypothesized that thyroid hormones play a role in the expression and function of ANGPTL4 in hepatocytes. We found that T3‐treated HepG2 and Hep3B cells showed increased ANGPTL4 mRNA and protein expression compared to vehicle treated cells. Interestingly, there was a concomitant decrease in the expression of the C‐terminal domain (37kDa) fragment of ANGPTL4 showing that T3 regulates not just the expression but also the posttranslational processing of ANGPTL4. Our results obtained in the human liver cells are contrary to findings in rodent livers. In conclusion, our studies suggest a probable mechanism by which T3 regulates lipoprotein levels. These studies were carried out using research support provided to Prabodh Sadana by Northeast Ohio Medical University.
The effect of nitric oxide on blockade of the norepinephrine transporter, uptake‐1, by imipramine
Authors: Simaan Joseph A.
Published: June 2021

In a previous study (Eur. J. Pharmacol, 2011) we showed that fresh synthesis of nitric oxide (NO) modulates transport of norepinephrine (NE) and tyramine (T) by uptake‐1 and its blockade by cocaine. We explored the effect of blocking the synthesis of NO with Nw‐nitro‐L‐arginine (L‐NNA) and restoration of the rise in pressure it induces with nitroglycerin, a nitric oxide donor (NG), on blockade of uptake‐1 by imipramine (IM), tested by modulation of the pressor responses of sympathomimetic amines that act directly like NE, that release NE like T, and which spare uptake‐1 like methoxamine (M), in anesthetized Sprague‐Dawley rats with measurement of mean arterial pressure. IM reduced the pressor effect of T by 81–85% which was restored after L‐NNA/NG by 233–641% (P<0.000) in different doses (0.025–0.1 mg). IM potentiated the pressor effect of NE by 60–86% (P<0.000), and decreased the pressor effect of M minimally with no further change after L‐NNA/NG. L‐NNA/NG relieved blockade of uptake‐1 and restored the pressor effect of T without further effect on NE because of opposing actions, relief of IM effect and direct depression of entry of NE. Minimal depression of the pressor effect of methoxamine is due to the alpha‐adrenoceptor blockade of IM. It is concluded that fresh synthesis of nitric oxide is significantly involved in the blocking activity of IM on uptake‐1. Supported by the American University of Beirut.
β‐Endorphin Inhibits the Expression of Amyloid Precursor Protein in SK‐N‐SH Neuroblastoma Cells expressing high levels of mutant APP
Authors: Boggeti Renuka , Bhatt Vrushank D , Ratna Warren N
Published: June 2021

Alzheimer's is a neurodegenerative disorder characterized by increased deposition of insoluble extracellular amyloid plaques in the brain with neurofibrillary tangles within the neurons. Amyloid plaque formation or amyloidosis results from the deposition of amyloid‐β peptide produced from amyloid precursor protein (APP) by the activity of γ‐secretase. APP gene promoter contains a cAMP response element (CRE) and is responsive to increase in intracellular cAMP levels. Agents that cause elevated intracellular cAMP through cell‐signaling have been shown to increase the expression of APP mRNA and protein levels. β‐endorphin has been shown to modulate intracellular cAMP through cell signaling. To determine the effect of beta‐endorphin on the expression of APP, SK‐N‐SH neuroblatoma cells were treated with β‐endorphin and its effect was studied using RNA and protein analyses. Northern blot and RT‐PCR analyses indicated decreased levels of transcription of the APP mRNA. Western blot analysis showed a corresponding decrease in the expression of the protein. Treatment with propranolol showed a similar effect; forskolin on the other hand caused an increase in APP mRNA levels and corresponding increase in APP expression. The elevated physiological release of β‐endorphin may likely have a beneficial effect in Alzheimer's patients. This work was supported by AMS College of Pharmacy, Long Island University, NY.
Interaction of eNOS and iNOS polymorphisms supports the role of genes related to nitric oxide (NO) bioavailability in migraine pathophysiology
Authors: Gonçalves Flávia Magazoni , Luizon Marcelo Rizatti , de Oliveira Alisson Martins , Speciali José Geraldo , Dach Fabíola , dos Santos José Eduardo Tanus
Published: June 2021

Migraine is a debilitating neurovascular disorder with high prevalence worldwide but its pathophysiology is not completely clarified. There is strong evidence that NO is critically involved in migraine etiology. In this regard, genes encoding for regulators of endothelial function and vascular permeability have been suggested as important candidate genes. However, although gene‐gene interactions are suggested to be an important component to complex disease, only a few studies focused on this issue in migraine. Therefore, we sought to characterize the possible interactions among polymorphisms of clinical relevance in the eNOS, iNOS and VEGF genes with migraine using multifactor dimensionality reduction (MDR) method. We observed a best model of interaction of the rs743506 SNP of eNOS gene with the G2087A SNP in exon 16 of iNOS gene associated when all migraine patients were compared to control group (C; N=103) (P=0.012). Interestingly, the same interaction best model was significantly associated when both migraine without aura patients (MWA; N=113) and with aura patients (MA; N=42) were compared to controls (P=0.030 and P=0.079, respectively). Despite the marginally significant MA result, the overall this finding suggests the role of genes related to NO bioavailability pathway in migraine and supports a similar pathophysiology basis for both disease phenotypes. Research support: CNPq, FAPESP and CAPES
Dual Ligand Complexes of Human Cytochrome P450 2B6 and Rabbit Cytochrome P450 2B4 with Amlodipine Reveal Substrate Access Channels into the Active Site
Authors: Shah Manish B , Stout C. David , Halpert James R
Published: June 2021

Substrate access channels have been predicted previously for several mammalian P450 enzymes based mainly on molecular dynamics simulations. We now report X‐ray crystal structures of human P450 2B6 and rabbit P450 2B4 in complex with two molecules of the calcium channel blocker amlodipine. The presence of the two drug molecules identifies clear access pathways in each P450. Based on the access channel nomenclature, 2f and 2a channels pointing towards the membrane correlate well with amlodipine access into the active site of P450 2B4. However, amlodipine appears to enter the active site of human P450 2B6 only via channel 2f. The results also indicate that key arginine residues located on the surface and at the entrance of the substrate access channels in each of the above P450s may play a crucial role in guiding substrate entry. In addition, secondary structure elements B′, F, F′, G′ and part of helix G were substantially retracted compared with the 4‐(4‐chlorophenyl)imidazole complexes of 2B4 and 2B6. Conformational variations were predominantly localized to the above elements that surround the substrate access channels as seen previously in the open P450cam structure. To our knowledge, the structures represent the first clear evidence from X‐ray crystallography of the substrate access channels into the active site in mammalian P450s. Supported by NIH grant ES03619.
Isoform‐selective changes in arachidonic acid (AA) metabolizing cytochrome P450 (P450) enzymes in warm hepatic ischemia (IR) reperfusion injury
Authors: Edpuganti Vindhya , Mehvar Reza
Published: June 2021

The objective of this study was to investigate the effects of warm hepatic IR injury on the major P450 enzymes responsible for epoxygenation (CYP2J2 and CYP2C23) and ω/ω‐1 hydroxylation (CYP4A) of AA in rats. Four groups of rats (n = 5–6/group) were subjected to 60 min of in vivo partial (70%) hepatic ischemia or sham operation, followed by 3 or 24 hr of reperfusion. Liver microsomes and plasma samples, collected at the end of the reperfusion period, were used for quantitation of alanine aminotransferase (ALT) concentrations and the protein levels of CYP4A, CYP2C23, and CYP2J2 (Western blot), respectively. Compared with sham‐operated animals, IR caused 10‐ and 3.3‐fold increases in the ALT concentrations after 3 and 24 hr of reperfusion, respectively (P<0.0001). In the 3‐hr reperfusion group, IR caused no changes in the expression of CYP4A and CYP2C23, whereas it down regulated CYP2J2 levels by 19.7% (P<0.01). In the 24‐hr reperfusion group, IR caused no changes in the expression of CYP2J2, whereas it upregulated CYP4A by 46.5% (P<0.05) and CYP2C23 by 56.1% (P<0.05). These results suggest that warm hepatic IR causes isoform‐selective changes in the AA‐metabolizing enzymes, potentially altering hepatic disposition of P450‐mediated metabolites of AA. Financial Support: Texas Higher Education Coordinating Board and Texas Tech School of Pharmacy Vascular Drug Research Center.