bluuebunny/crossref_metadata_2025_split huggingface dataset:
Repeated Non‐invasive Remote ischemic Preconditioning Confers Cardioprotection in Diabetic Rats (LB559)
Authors: Wang Chunyan , Li Haobo , Xia Zhengyuan , Irwin Michael 
Published: June 2021

Non‐invasive Remote ischemic preconditioning (NRIPC) has a cardioprotective effect in normal rats, but its effect in rats with diabetes is controversial. This study was mainly to determine the cardioprotective effect of NRIPC on ischemia reperfusion injury (IRI) in diabetic rats. Diabetes was induced in Sprague‐Dawley rats with streptozotocin. NRIPC was implemented by applying 3 cycles of 5‐min occlusion/5‐min reperfusion in a unilateral hindlimb once per day for three days. The animals were then subjected to 30 minutes of coronary artery occlusion followed by two hours of reperfusion. A control group had no NRIPC. The results showed that NRIPC significantly reduced post‐ischemic myocardial infarct size,lactate dehydrogenase (LDH) and troponin‐1 release during reperfusion in both control and diabetic rats. Protein PKC‐ε, phosphorylation of AKT and signal transducer and activator of transcription 3(STAT3) were all significantly increased after NRIPC and their enhancement after reperfusion was correlated with reductions of post‐ischemic myocardial infarction and cellular injury. In conclusion, cardioprotection can be achieved in diabetes by NRIPC applied repeatedly for three days prior to inducing myocardial infarction and activation of PKC‐ε and STAT3 might be a major mechanism of NRIPC protection.Grant Funding Source: RGC/GRF grant (768211M) and a HKU seeding grant
Effect of scutellarin on JAK/STAT pathway in myocardial ischemia reperfusion rats (LB561)
Authors: Yang Weimin , Li Lin , Cai Donghua , Li Jiaxun , Du Yue , Guo Tao 
Published: June 2021

Animal studies have demonstrated that scutellarin (SCU) limits damage after myocardial ischemia injury. However, the underlying molecular mechanisms of SCU protecting against myocardial injury induced by ischemia reperfusion (IR) are not well known. This study examined whether SCU protects against myocardial IR injury in rats is mediated by the Janus kinase and signal transducer and activator of transcription (JAK/STAT) pathway. Protein expression of JAK2/STAT3 and phosphorylative products were assessed by Western blot and immunohistochemistry methods in myocardial tissue from myocardial IR rats pretreated with SCU (45 and 90 mg/kg, iv). Western blot and immunohistochemistry assays showed that myocardial IR injury caused significant increases of phosphorylated JAK2 (p‐JAK2) and phosphorylated STAT3 (p‐STATS) levels in the rat myocardial IR model. SCU (45 and 90 mg/kg, iv) significantly reduced ischemic size, and decreased histological p‐JAK2 and p‐STATS expression. SCU attenuates myocardial IR injury which involved in JAK/STAT signaling pathway.Grant Funding Source: Supported by grants from the National Natural Science Foundation of China (Nos. 30960450 and 81173110) and Yunnan Provincial Science and Technology Department (Nos. 2011FA022, 2012BC012, 2008CD054, and 2008ZC111M).
Functional overexpression and purification of a codon optimized synthetic four superantigens in Escherichia coli: Production of superantigens with improved safety and efficacy for cancer immunotherapy (LB582)
Authors: Bashraheel Sara , Al‐Qahtani Alanod , Al‐Sulaiti Haya , Al‐thani Amna , Goda Sayed 
Published: June 2021

Bacterial superantigens (SAGs) are potent T cell stimulatory molecules and comprise a large family of disease‐associated proteins. The best‐characterized members of the superantigen family are staphylococcal enterotoxins (SEs), which are secreted by Staphylococcus aureus, and streptococcal pyrogenic exotoxins (SPEs), which are secreted by Streptococcus pyogenes Unlike conventional antigens they are not processed internally by antigen presenting cells (APC), and are thus not displayed as peptide antigen in the peptide‐binding groove of the MHC class II molecule. Superantigens bind to APCs on the outside of the MHC class II molecule and to T cells via the external face of the T cell receptor (TCR). This enables them to activate up to 20% of resting T cells, whilst conventional antigen presentation results in the activation of 0.001‐ 0.0001% of the T cell population. These biological properties of superantigens make them very attractive for use in immunotherapy.In this study, four synthetic genes coding for four different superantigens, SEA, SEB, TSST‐1 and SPEA were codon‐optimised and synthesized for maximum expression in E. coli using the vector pET28a.Our work indicated that in all cases the recombinant superantigens were expressed to 40% of the total host protein. We managed to optimise the condition to obtain 20% of the recombinant superantigens in a soluble form. The purification of the soluble His‐tagged recombinant superantigens have been achieved in a single step by Ni2+ charged column chromatography.The superantigenicity of each synthetic recombinant SEA, SEB, TSST‐1 and SPEA was determined by 2 methods. First method includes 3H‐thymidine incorporation assay as a measure of T‐cell proliferation i.e. T‐cell stimulation by modified SAGs. The second part of the superantigenicity assays includes measurement of cytokines produced by peripheral blood cells.The successful cloning and expression of four codon optimised synthetic superantigens genes will provide the recombinant protein for production of safe superantigen for cancer immunotherapy.Grant Funding Source: Anti‐doping lab‐Qatar
Use of Levitated Cells n3D BiOAssay System to Determine Cytotoxicity of Calcium Sulfide (CaS) Clusters in Malignant Cells as Potential Cancer Cell Targeted Therapy (LB583)
Authors: Carrasco Nestor , Ortiz Carlos , Quiñones Jaileen , Feliciano Geishamarie , Ortiz Angel , Rivera Daniel , Castro Miguel , Suarez Edu 
Published: June 2021

Preliminary results in our lab showed that naked Calcium Sulfide (CaS) nanoparticles (5.0nm) decreased cell proliferation in the carcinoma cell lines CRL‐2124. Objective: The objective of this study was to determine if CaS clusters measuring < 3.0 nm were cytotoxic to malignant cells when compared to benign fibroblasts CRL‐2522 by using the n3D BiOAssay (Biosciences, Inc.). Methods: A total of 100,000 cells per cell line were seeded in T‐75 flasks. When reaching 80% confluence, they were incubated with the n3D nano‐shuttles overnight (ON) at 37oC, 5% CO2, and 98% relative humidity. Next day, cells were trypsinized, counted, and seeded (1.4 ‐ 1.6 M/per well) in 6‐well plates using the n3D BiOAssay to levitate the cells. A total of 150,000 cells/well were seeded in a 96‐well plate in triplicates with a total volume of 300ul (150ul of cells, 120ul of media, and 30ul of the treatments). The treatments consist of 10 pmoles of CaS, 2% DMSO, or growth media. Analysis for cytotoxicity were performed with the ImageJ software by measuring the “O” ring outer and inner diameters. Results: The cell were successfully levitated by the n3D BiOAssay System. The expected “O” ring was formed before the treatments were added. We observed changes in the diameters for the CaS treated wells. Conclusions: The n3D BiOAssay provides an excellent and cost effective system for testing cellular toxicity of compounds with the potential to be used as therapeutic agents. We found that malignant cells treated with CaS showed more cytotoxicity than benign normal cells. CaS clusters may represent a novel Cancer Cell Targeted Therapy.Grant Funding Source: NIH‐NIGMSR25GM096955, NIH‐R25GM088023
The combination of curcumin and ginkgolide B reduces cyst growth by multiple cell signaling pathways (LB586)
Authors: Gao Jinsheng , Li Yousong , Wang Xixing , Hao Xuliang , Liu Likun , Zhou Hong , Yang Baoxue 
Published: June 2021

Autosomal dominant polycystic kidney disease (ADPKD), a common disease with high incidence ratio, often causes great harm to people. However, no effective drug is currently available. The purpose of this study was to evaluate the cyst coinhibition of curcumin and ginkgolide B and to elucidate their different inhibition mechanisms. In vitro MDCK cyst model was used to observe the coeffect of curcumin and ginkgolide B on cyst development. The results showed that the shapes of cysts were different after curcumin or ginkgolide B treatment. The cysts treated with curcumin often became small thick‐walled cysts, the cysts exposed to ginkgolide B often turned into small thin‐walled cysts, and the cysts after coinhibition of curcumin and ginkgolide B often became round shaped colonies, which suggested the combination of two drugs was more effective than any of them. Our study and foreign study have confirmed the cyst inhibition effect of curcumin and ginkgolide B on polycystic kidney disease mice. Curcumin may execute its anti‐cyst activity partly by blocking MAPK signaling pathway and m‐TOR signaling pathway. Furthermore, our study demonstrated that curcumin up‐regulated Ca2+ level in MDCK cell, which may be related to its cyst inhibition. Ginkgolide B may execute its anti‐cyst activity partly by blocking MAPK signaling pathway. The results provide proof‐of‐concept for application of combination of curcumin and ginkgolide B to reduce cyst growth in polycystic kidney disease.Grant Funding Source: The National Youth Science Fund (81302828), Shanxi Province Youth Science and Technology Research Fund (2013021037‐3), Taiyuan Science and Technology Research Fund (201110‐1555), Shanxi Province Youth Science and Technology Research Fund (201201040)
Inhibition of Buprenorphine Metabolism by Proton Pump Inhibitors: Enigmatic Time‐Dependent Effect of Esomeprazole (LB594)
Authors: Moody David , Liu Fenyun , Fang Wenfang 
Published: June 2021

While several studies have investigated the effect of proton pump inhibitors (PPI) on the activities of specific cytochrome P450s (CYP450), most studies used model substrates. Here we investigate the effect of PPIs on the N‐dealkylation of buprenorphine, a CYP3A4 and 2C8 driven reaction. An initial screen was performed using three concentrations of PPI in human liver microsomes (HLM) with or without a 15‐minute pre‐incubation of the HLM with inhibitor and a source of NADPH. For five PPIs, the following apparent IC50s (µM) were determined without (‐) or with (+) 15 minute pre‐incubation: omeprazole, (‐) 20.9, (+) 2.5; esomeprazole, (‐) 9.8, (+) < 1.0; lansoprazole, (‐) > 100, (+) > 100; pantoprazole, (‐) 120, (+) 16.2; and rabeprazole (‐) 79.4, (+) 60.3. For all but lansoprazole, pre‐incubation effects suggest time‐dependent inhibition (TDI). Subsequent studies with cDNA‐expressed CYPs (rCYP) were conducted with 6 or more concentrations of inhibitor to determine an IC50. Respective IC50s (µM) determined with rCYP3A4 and 2C8 were: omeprazole, 220 and 65; esomeprazole, 90 and 65; lansoprazole, > 100 and 90; pantoprazole, > 200 and 200; and rabeprazole, 150 and 70. TDI tests were also conducted for specific PPIs with a dual incubation system. From a primary incubation with 3 or more inhibitor concentrations, aliquots then transferred (and diluted) after varying pre‐incubation times to a secondary system with buprenorphine and additional NADPH. These were conducted for omeprazole, esomeprazole, lansoprazole, pantoprazole and rabeprazole with rCYP3A4, and omeprazole and esomeprazole with rCYP2C8. Only rabeprazole produced pre‐incubation time‐ and inhibitor concentration‐dependent decreases in CYP3A4 activity (Ki = 2.6 µM, kinact = 0.052 min‐1). None of the others demonstrated TDI, thus an enigma arose between HLM and rCYP TDI results. When we performed the dual incubation using esomeprazole and HLM, inhibition was pre‐incubation time‐ and inhibitor concentration‐dependent (Ki = 20.9 µM and kinact = 0.0265 min‐1). Therefore, we hypothesize that metabolism of esomeprazole (and likely omeprazole and pantoprazole) by another CYP produces a potent inhibitor for CYP3A4 and/or CYP2C8 metabolism of buprenorphine.Grant Funding Source: Supported by National Institute of Justice Award #: 2011‐DN‐BX‐K532
Central administration of C‐X‐C chemokine receptor type 4 antagonist alleviates the development and maintenance of peripheral neuropathic pain in mice (LB618)
Authors: Luo Xin , Tai Wai , Sun Liting , Qiu Qiu , Xia Zhengyuan , Chung Sookja , Cheung Chi Wai 
Published: June 2021

C‐X‐C chemokine receptor type 4 (CXCR4) is an emerging area of pain hypersensitivity in the peripheral nervous system; however, its role in the central nervous system (CNS) for pain processing remains unclear. This study aimed at exploring the function of central CXCR4 in pain processing in vivo using its specific antagonist AMD3100 and partial sciatic nerve ligation (pSNL) model of peripheral neuropathic pain in C57BL/6 mice. A single intrathecal (central) administration of AMD3100 (intrathecal AMD3100, 5 μg) was found to prevent the development of pSNL‐induced allodynia. Intrathecal AMD3100 (1 μg, 5 μg and 25 μg) also reversed the established pSNL‐induced mechanical allodynia in a dose‐dependent way on post‐operative day 7 (POD 7). In rotarod test, intrathecal AMD3100 of different dosages (1 μg, 5 μg and 25 μg) did not impair the motor function of mice. Among mitogen‐activated protein kinases (MAPKs) pathways, intrathecal AMD3100 was found to downregulate the activation of JNK1 and p38 pathway signaling proteins in the L3‐L5 spinal cord segment as assessed by western blotting on POD 7. Our results firstly suggest that central (spinal) CXCR4 appears to be involved in the development and maintenance of peripheral neuropathic pain, implicating that CXCR4 would potentially be a therapeutic target for chronic neuropathic pain.Grant Funding Source: Supported by the departmental fund, Department of Anaesthesiology, The University of Hong Kong
Cocaine induces HIF1 (Hypoxia Inducible Factor1) in rat C6 glial cells (LB619)
Authors: Agharahimi Maryam , Goodman Carl , Mazzio Elizabeth , Taka Equar 
Published: June 2021

Investigations with astroglial cells carry more prominence in drug abuse studies. However, not many investigations have been conducted on the toxicity of various abused drugs, like cocaine. The present study was aimed to discern the effect of cocaine on rat astroglioma cells. Glial cells were grown in 10% FBS (Fetal Bovine Serum) in Dulbecco’s modified Eagle medium (DMEM) and penicillin/streptomycin (100 Units/0.1 mg per ml). Treatment of glial cells with cocaine for 24 and 48 hours caused significant loss of cell viability. We also used rat cytokine array membranes to determine the change in rat C6 glial cell cytokine expression upon cocaine exposure (2, 4 and 7 mM). We found after 24 hours cocaine exposure, expression of vascular endothelial growth factor (VEGF) was relatively high in each concentration compared to control and these results were confirmed with western blot analysis. Preliminary data from different studies indicated that hypoxia is a potent inducer of VEGF in vitro partly because of an increased transcription rate, mediated by binding of hypoxia‐inducible factor‐1 (HIF1) to a hypoxia responsive element, VEGF gene. Thus following cocaine treatment of various concentration and RNA isolation, real time PCR (Polymerase Chain Reaction ) was carried out using the IQ Sybr Green Supermix in conjunction with MyiQ software (Bio‐Rad, Hercules, CA). Expression of HIF1 followed by VEGF observed in each concentration compared to control. We concluded that cocaine may be involved in hypoxia in C6 glial cells.Grant Funding Source: This research was supported by National Institute on Minority Health Disparities of the National Institute of Health under award number G12MD007582, FAMU RCMI, Pharmaceutical Research Center
Interactions of APOE genotype and RXR agonists on soluble Aβ and oAβ levels (LB624)
Authors: Koster Kevin , Luo Jia , Lee Sue , Shete Varsha , Thatcher Gregory , LaDu Mary Jo , Tai Leon 
Published: June 2021

Interactions of APOE genotype and RXR agonists on soluble Aβ and oAβ levels Kevin Koster1, Jia Luo2, Sue Lee2, Varsha Shete2, Gregory RJ Thatcher2, Mary Jo LaDu1, Leon M Tai1 There is a critical need to develop novel Alzheimer’s disease (AD) therapeutics that lower amyloid beta (Aβ) levels, as current therapeutics are failing in clinical trials. APOE4 (encodes apolipoprotein E; apoE) is the greatest AD genetic risk factor compared to APOE3. ApoE represents a promising target for modulating A levels, as it has been shown to have genotype‐specific effects on Aβ pathology in humans and mice. Recently, bexarotene (Bex), a retinoid X receptor (RXR) agonist, was shown to reduce soluble and insoluble levels of Aβ via increasing mouse apoE levels. However, subsequent data have raised concerns over the activity and mechanism of action of Bex. A major issue is that the effect of modulating human apoE on Aβ levels is unclear. Further, the levels of Aβ pathology at time of treatment may determine RXR agonist activity i.e. treatment vs prevention paradigms. Therefore, novel EFAD mice (overexpress Aβ42 and express human apoE3 or apoE4) were treated with Bex and LG268, a more selective RXR agonist. RXR agonist treatment of E3FAD and E4FAD mice with intermediate Aβ levels resulted in increased soluble Aβ42 and oligomeric Aβ levels (oAβ), with a trend to increase insoluble Aβ levels. RXR agonist treatment of E4FAD mice with high Aβ levels decreased soluble Aβ42 and oAβ levels, with no effect on insoluble Aβ. Further, our data indicate that the beneficial effects of RXR agonists are mediated through: increased ABCA1 and ABCG1 expression, increased lipidation of apoE‐containing lipoproteins, and thus increased apoE/A complex levels. Collectively these data suggest that RXR agonists may increase neurotoxic Aβ levels in prevention paradigms. However, RXR agonists may be effective for AD patients expressing APOE4 at a more advanced stage of the disease via a novel mechanism of action: enhanced apoE4 lipidation.Grant Funding Source: Supported by Alzheimer's Disease Drug Discovery Foundation
Targeting cathepsins‐tBid‐mitochondial apoptotic signaling pathway in astrocytes contributes to neuroprotection against cerebral ischemia (LB627)
Authors: Zhang Hui‐Ling , Xu Min , Yang Lei , Rong Jia‐Guo , Ni Yong , Ishidoh Kazumi , Katunuma Nobuhiko 
Published: June 2021

ABSTRACT The roles of cathepsins in the ischemic astrocytic injury remain unclear. Here we test the hypothesis that activation of cathepsin B and L contributes to the ischemic astrocyte injury via the tBid‐mitochondrial apoptotic signaling pathways. In the rat models of pMCAO, CA‐074Me or Clik148, a selective inhibitor of cathepsin B or cathepsin L, reduced the infarct volume, improved the neurological deficits and increased the MAP2 and GFAP levels. In OGD‐induced astrocyte injury, CA‐074Me or Clik148 decreased the LDH leakage and increased the GFAP levels. In the ischemic cortex or OGD‐induced astrocytes injury, Clik148 or CA‐074Me reversed pMCAO or OGD‐induced increase in active cathepsin L or cathepsin B at 3 h or 6 h, increase in tBid, reduction in mitochondrial Cyt‐c and increase in cytoplastic Cyt‐c and active caspase‐3 at 12‐24 h of the late stage of pMCAO or OGD. CA‐074Me or Clik148 also reduced cytosolic and mitochondrial tBid, increased mitochondrial Cyt‐c and decreased cytoplastic Cyt‐c and active caspase‐3 at 6 h of the early stage of Bid activation. CA‐074Me or Clik148 blocked the pMCAO‐induced release of cathepsin B or L from the lysosomes into the cytoplasm and activation of caspase‐3 in ischemic astrocytes at 12 h after ischemia. Concurrent inhibition of cathepsin B and cathepsin L provided better protection on the OGD‐induced astrocytic apoptosis than obtained with separate use of each inhibitor. These results suggest that inhibition of the cysteine cathepsin B and cathepsin L activation in ischemic astrocytes contributes to neuroprotection via blocking the tBid‐mitochondrial apoptotic signaling pathway. This work was supported by grants from The National Natural Science Foundation of China (30973510, 81171104, 30930035).Grant Funding Source: This work was supported by grants from The National Natural Science Foundation of China (30973510,
Dexmedetomidine suppresses gap junctional intercellular communication and attenuates the sensitivity of gliocytoma to temozolomid (LB642)
Authors: Wang Lingzhi , Zhang Suzhi , Huang Huansen , Xia Weiyi , Xia Zhengyuan 
Published: June 2021

Dexmedetomidine(DEX), an alpha‐2 adrenergic agonist which can affect cell‐to‐cell gap junctions(GJ), is used as an adjuvant analgesic and sedatives in patients with cancer pain, especially in neural cancer patients who concomitantly receiving chemotherapy. GJ may increase the sensitivity of tumor cells to chemotherapeutic agents (Oncol Rep. 2014;31:540‐550). It is unknown whether or not DEX may affect the sensitivity of gliocytoma (U87 cell, which richly expresses the GJ protein Connexin43) to temozolomide (TEM, an anticancer agent for gliocytoma) and if DEX exerts its effect via affecting GJ. U87 cells were treated with TEM, respectively at high density (which form GJ) or low density (without GJ formation), for 1 hour in the absence or presence of 3 hours DEX pretreatment prior to applying TEM. TEM toxicity (i.e., reduction of clonogenic cell survival), was assayed by “Standard Clony‐forming assay” and GJ function was examined by “Parachute” dye‐coupling assay. TEM toxicity was greater at high density than at low density cells (P<0.01), while either oleamide (a GJ inhibitor) or Cx43 siRNA reduced TEM toxicity. Similarly, DEX reduced GJ function and compromised TEM treatment effects, manifested as increases in clonogenic cell survivals at high but not at low cell density. We concluded that DEX reduced TEM cytotoxicity through inhibiting GJ function in gliocytoma.Grant Funding Source: supported by departmental development fund and Supported by Science and Technology Planning Project of Guangdong Province(2010B060900089)
Angiotensin‐(1‐7) infusion in the brain reduced blood pressure and cardiac hypertrophy in (mRen2)27 transgenic hypertensive rats (LB647)
Authors: Kangussu Lucas , Guimaraes Priscila , Nadu Ana Paula , Bader Michael , Santos Robson , Campagnole‐Santos Maria 
Published: June 2021

Ang‐(1‐7) through Mas receptor is involved in the control of cardiovascular function and may present important cardioprotective actions after peripheral administration. In the present study, Sprague Dawley (SD) and transgenic hypertensive rats (mRen2)27, instrumented with telemetry probes for arterial pressure (AP) measurements, were subjected to 14 days of lateral ventricle (ICV) infusion of Ang‐(1‐7) (200 ng/h) alone, Ang‐(1‐7) (200 ng/h) in association with a Mas receptor antagonist (A779, 1 μg/h) or saline (0.5 µl/hour) through osmotic mini‐pumps. ICV Ang‐(1‐7) attenuated the hypertension of (mRen2)27rats and this effect was completely blocked by A779. The AP lowering effect of Ang‐(1‐7) was accompanied by a significant improvement of the baroreflex control of heart rate and an attenuation of the increased ratio of sympathetic/vagal activity to the heart of (mRen2). Further, there was a significant attenuation of the increased levels of atrial natriuretic peptide, brain natriuretic peptide, collagen I, fibronectin, TGF‐β1 and hypertrophy in the heart of (mRen2)27 rats. All effects induced by ICV Ang‐(1‐7) were reduced by the concomitant ICV infusion of A‐779. These data indicate that increased levels of Ang‐(1‐7) in the brain may contribute to the treatment of hypertension, not only by reducing the AP, but also leading to an attenuation of cardiac remodeling, at least in (mRen2) 27 hypertensive rats. These effects of Ang‐(1‐7) are likely to be mediated by Mas receptor interaction in the brain.Grant Funding Source: Supported by CAPES, FAPEMIG and CNPq through INCT NanoBioFar and PRONEX.