bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of novel in vitro and in vivo models to evaluate antibiotic efficacy against shigellosis
Authors: Arnold Samuel , McCloskey Molly , Shaheen Shareef , Hulverson Matthew , Choi Ryan 
Published: June 2021

Shigellosis is caused by oral ingestion of the gram‐negative bacteria Shigella. Symptoms of the disease include severe diarrhea and dysentery, and the impact of the disease disproportionately affects children and those in low‐income countries such as sub‐Saharan Africa, Asia, and Latin America. According to a reanalysis of the Global Enteric Multicenter Study (GEMS), Shigella has the highest attributable incidence of pathogen specific moderate to severe diarrhea in children between 12–59 months of age. Recent studies have estimated approximately 90 million shigellosis cases annually, and the Global Burden of Disease 2015 analysis suggests at least 164,000 deaths per year are due to Shigella. Interspecies‐variation with lack of cross‐protection may challenge the development of a broadly‐protective vaccine and no human vaccine candidates are currently approved for treatment. Therapeutic options for shigellosis are limited and globally emerging drug resistance to sulfonamides, tetracyclines, ampicillin, and TMP‐SMX has been reported. With resistance to many “first‐line” antibiotics, ciprofloxacin, azithromycin, and ceftriaxone have been recommended for treatment of shigellosis. However, rapidly evolving resistance has been reported for these antibiotics as well. While there is a clear need for novel therapeutics to treat this disease, current drug development efforts are hindered by a lack of fundamental in vitro and in vivo tools to assess the efficacy of pre‐clinical candidates. We have focused our efforts on establishing in vitro co‐culture systems and in vivo mouse models to evaluate small molecule therapeutics. Our aim is to use these tools to identify PK/PD relationships for established and novel therapeutics to treat Shigella infection. The gastrointestinal localization of the parasite generates a unique challenge for establishing a pharmacokinetic‐pharmacodynamic (PK/PD) relationship for an anti‐Shigella drug candidate. We have previously reported on our use of physiology based pharmacokinetic (PBPK) modeling to identify an association between gastrointestinal drug concentrations and in vivo efficacy for anti‐Cryptosporidium drugs and we plan to use a similar technique for the antibiotics used to treat Shigella infection. The presented work will be valuable for the entire research community currently engaged in the development of therapeutics for enteric diseases.Support or Funding InformationThis work was supported by the Tres Cantos Open Lab Foundation Award: TC246This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Trends and Advances on Precision Medicine through Scientometric Analysis
Authors: Sohn Eunsoo , Sohn Eun‐Hwa 
Published: June 2021

Precision medicine, more advanced concepts of personalized medicine, triggered by United States' Precision Medicine Initiative has become a new paradigm of health care. Countries around the world are planning to develop precision medicine as a national strategic project, and collaborative research is being actively carried out centered on developed countries. They are making various efforts to drive toward precision medicine by establishing a large scale clinical cohort studies, etc. Despite some current limitations, many opportunities and challenges still remain in precision medicine. In this paper, we reviewed recent R&D status and trends in precision medicine by scientometric analysis methods. Total 19,403 articles retrieved from the scopus database have analyzed using KnowledgeMatrix Plus, Gephi and VOSviewer software. Especially each nation's activities and their technology position, international collaboration and hot research area in precision medicine have also been analysed and identified through network and co‐word analysis showed by visualization map. Results showed that the R&D activities on precision medicine had a dramatic increase in recent years and were located in the early stage of technology development with differentiation by country.Support or Funding InformationThis research was supported by Korea Institute of Science and Technology Information (KISTI).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Phospholipids as Indicators of Castration Resistant Prostate Cancer
Authors: Ingram Lishann M. , Manosoura Maryam , Pati Sumitra , Cummings Brian 
Published: June 2021

The association of circulating lipids with clinical outcomes of metastatic castration‐resistant prostate cancer (CRPC) is not fully understood. Interestingly, recent studies demonstrated a correlation between plasma lipids and CRPC prognosis, resulting in a proposed prognostic four‐lipid signature of (ceramide, sphingomyelin, phosphatidylcholine (PC) and phosphatidylserine (PS)). While the increase in these lipids correlated to decreased survival, the mechanisms mediating alterations in these lipids, and the correlation to resistance to treatment are not known. We addressed this gap‐in‐knowledge using in vitro models of non‐cancerous, hormone sensitive and CRPC cell lines and quantitative lipidomic analysis. An initial untargeted approach assessing both cells and media indicated that ceramides, sterols and glycerophospholipids had higher relative abundance in CRPC cells (PC‐3 and DU145), as well as hormone sensitive cells (LNCaP), as compared to a non‐cancer cell model (PNT2). Similar data was seen in media. A targeted approach focusing on lipid species shown to correlate to survival in recent publications showed significantly higher levels of 36:2 phosphatidylserine (PS) in all prostate cancer cell lines tested, and lower levels of 36:2 PC in the media of these same cells. Thus, both untargeted and targeted analysis of prostate cancer cells showed significant differences in lipidomic profiles as compared to non‐cancer prostate cells. These data suggest that the lipidomic profiles of prostate cancer cells lines mirror lipidomic profiles in the plasma of prostate cancer patients. As such, these cells may prove useful for understanding the molecular mechanism mediating lipid changes in prostate cancer patients and yield insight into the role of lipids in mediating CRPC.Support or Funding InformationNational Institute of Biomedical Imaging and Bioengineering NIBIB (EB0160100)Department of Defense Prostate Cancer Research Program Idea Development Award (PC150431 GRANT11996600)This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Relative Neutralization of Heparin from Different Origins by Protamine, Polybrene, Platelet Factor 4, and synthetic heparin antagonist PMX
Authors: Laddu Rajan , Farooqui Ambar , Siddiqui Fakiha , Iqbal Omer , Hoppensteadt Debra , Kouta Ahmed , Fareed Jawed 
Published: June 2021

IntroductionHeparin is the most widely used anticoagulant, which is derived from mammalian tissues. While porcine mucosal heparin is most commonly used clinically, sheep mucosal heparin and bovine mucosal heparin are also currently developed. Protamine Sulfate is the most commonly used antagonist of heparin, which is used clinically. Platelet Factor 4 is a platelet‐derived protein which is capable of neutralizing heparin endogenously. Polybrene and PMX are synthetic agents which can neutralize heparin. These agents are not used clinically. The purpose of this study is to compare the neutralization profile of various heparins by protamine and other heparin‐neutralizing agents.Materials and methodPorcine heparin was obtained from Medefill Inc. (Glendale Heights, IL). Bovine Heparins were obtained from Extrasul (Sao Paolo, Brazil). Ovine Heparin was obtained from Ronssi Co. (Souzhou, China). Protamine sulfate was obtained from Sanofi Avantis Co. (Paris, France). Platelet Factor 4 was obtained from Enzyme Research (South Bend, IN). Polybrene and PMX were of synthetic origin. Pool‐citrated plasma was obtained from Loyola University Medical Center Blood Bank. Blood‐clotting tests were performed using aPPT and TT reagents. Chromogenic substrate assays were carried out on the ACL‐Elite instrument. Plasma samples were supplemented with each of the pooled heparin preparations at a concentration of 0–10 mg/ml. Protamine neutralization studies were carried out at a fixed concentration of 10 mg/ml. Neutralization profiles were determined graphically and by calculating percent neutralization at a fixed concentration of 5 mg/ml.ResultsAt 5 ug/ml the protamine, the neutralization index for the different heparins were comparable in the TT, anti‐Xa and anti‐IIa assays (> 90%) however in the APTT it was around 75% for porcine and bovine pool and 66% for ovine pool. In the polybrene neutralization the all heparins comparable neutralization index in the APTT (>80%), TT (>90%) and anti‐IIa (>90%) however in the anti‐Xa the porcine and bovine were greater than 80% and bovine pool was greater than 90%.ConclusionsProtamine sulfate was found to be the most effective neutralizing agent for the bovine, ovine and porcine heparins. Assay based variations were noted in the neutralization profiles. With polybrene and platelet factor 4. These studies suggest that titration with the heparin neutralizing agent may be useful in demonstration the biosimilarities of heparins obtained from different sources.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The association of Vitamin D levels to advanced liver fibrosis and response to hepatitis C therapy
Authors: Elhag mashair Khougali Abdelrahman , Deiab shihab , Hussain Nadia , Aziz Rasha Khougali Abdelrahman 
Published: June 2021

The importance of vitamin D in many diseases has come to light recently. In the Middle East, there is a prevalence of low vitamin D levels. Evidence linking vitamin D to the parthogenesis of chronic hepatitis C is present. The aim of this study is to analyze the relationship of vitamin D levels with advanced liver fibrosis (ALF) CHC treatment‐naive patients and sustained virology response (SVR) in CHC patients on PEGylated interferon alpha plus ribavirin (pegIFNa/ribavirin) therapy.Materials and methodsA meta‐analysis of eligible studies published from 1999 to 2014 was conducted. Databases such as PubMed, SCOPUS, Web of Science, and the Cochrane Library were used. Studies assessed included data assessment between plasma/serum vitamin D levels and its relation to ALF and/or SVR. Pooled odds ratios (ORs) were estimated by either fixed or random effects models. Eighteen studies were selected from the literature search, seven for ALF (1,099 patients) and 11 for SVR (2,580 patients). For liver fibrosis, low vitamin D status was related to a diagnosis of ALF, with the cutoffs of 10 ng/mL (OR=2.37, 95% confidence interval, [CI] = 1.20, 4.72) and 30 ng/ml (OR=2.22, 95% CI = 0.99, 3.97) being significant, and a near‐significance for 20 ng/ml.ResultsRegarding SVR, there was a significant heterogeneity among studies (P< 0.001). A significant association with SVR for a vitamin D cutoff of 20 ng/mL (OR=0.53, 95% CI=0.31, 0.91) was present. When meta‐analysis was performed excluding the outliers, significant pooled ORs were found for all patients (10 ng/mL [OR=0.48, 95% CI=0.34, 0.67] and 20 ng/mL [OR=0.58, 95% CI=0.45, 0.76]) and GT1/4 patients (10 ng/mL [OR=0.53, 95% CI=0.34, 0.81] and 20 ng/mL [OR=0.54, 95% CI=0.39, 0.74]).ConclusionLow vitamin D status in CHC patients is associated with a higher likelihood of having ALF and lower odds of achieving SVR following pegIFN/ribavirin therapy. Also vitamin D supplementation could improve the SVR rate in patients treated with pegIFN/ribavirin.Support or Funding InformationThis work was supported and funded by Ras Alkhaimah medical and health science universityThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A novel metformin‐methylglyoxal imidazolinone metabolite (IMZ) sensitizes cells to insulin; a potential role in alleviating T2DM complications
Authors: Hargraves Tiffanie L. , Mastrandrea Nicholas J. , Lau Serrine S. , Monks Terrence J. 
Published: June 2021

Reactive dicarbonyls, such as methylglyoxal (MG), are elevated in type‐two diabetes mellitus (T2DM) patients. These endogenous electrophiles covalently modify proteins, which may contribute to diabetic complications. The T2DM first‐line therapy, metformin (MF), significantly reduces adverse diabetic endpoints and mortality more effectively than other anti‐hyperglycemic agents, the mechanism(s) of which remain unclear. We previously identified and characterized the product of the MF and MG reaction as a novel five‐membered imidazolinone (IMZ) metabolite. IMZ was detected via LC/MS in all MF‐treated T2DM patients, and increased MF urinary levels directly correlated with elevations in urinary IMZ. Scavenging of MG by MF represents a possible alternative mechanism of MF drug efficacy, in addition to its antigluconeogenesis properties. Imidazoline receptor subtypes (I1R, I2R, and I3R) are novel targets for drug development in disorders associated with T2DM because they are involved in insulin sensitization, insulin secretion, and glucose homeostasis. Thus, we examined the ability of IMZ to modulate insulin‐mediated cell signaling pathways via western blot in PC12 cells, which express high levels of I1R and lack the α2‐adrenergic receptor, which many I1R agonists also activate. Combination treatment of insulin and IMZ at physiologically relevant concentrations increased AKT and ERK1/2 phosphorylation above levels seen with insulin treatment alone. This potentiation was not observed in the presence of I1R antagonists. Moreover, IMZ restores insulin sensitivity in an insulin resistant HEPG2 cell model. The results revealed that IMZ may enhance insulin action in the insulin‐dependent AKT and ERK1/2 pathways through I1R activation. Preliminary in vivo pharmacokinetic studies show that IMZ (IP; 10mg/kg and 20mg/kg) is rapidly absorbed, with an elimination half‐life of 12–16.5 minutes. In addition, IMZ enhanced hepatic pAKT 40 minutes after the 10mg/kg dose. In summary, the formation of IMZ in T2DM patients provides evidence that MF scavenges MG, potentially reducing detrimental protein modifications. This property of MF may play a role in the reduction of diabetic complications and represent a potential alternative mechanism of MF drug efficacy. Further, IMZ itself may have the ability to enhance insulin action, suggesting possible clinical utility in the treatment of T2DM.Support or Funding InformationR24DK0090958, T32ES007091, P30ES006694This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The oxidized phospholipid POVPC induces endothelial to mesenchymal transition
Authors: Ou Zhi‐Jun , Li Yan , Chen Jing , Li Shang‐Xuan , Wang Tian‐Tian , Shi Mao‐Mao , Ou Jing‐song 
Published: June 2021

The exact mechanisms of atherosclerosis remian unclear. Endothelial to mesenchymal transition (EndMT) plays an important role in atherosclerosis. 1‐Palmitoyl‐2‐(5‐oxovaleroyl)‐sn‐glycero‐3‐phosphocholine (POVPC), the oxidation product of oxidized low‐density lipoprotein, is an oxidized phospholipid and has been found in atherosclerotic lesions. We recently demonstrated that POVPC could impair endothelial function and induce endothelial cell apoptosis. However, the effects of POVPC on EndMT are unclear. Human aortic endothelial cells were treated with POVPC for long term (at least 48 h). Endothelial cell viability and morphology were observed. Nitric oxide (NO) production and superoxide anion generation (O2•−) were measured. The expression of specific endothelial markers cluster of differentiation 31 (CD31) and mesenchymal markers alpha smooth muscle actin (α‐SMA) were detected by immunofluorescence and immunoblotting. The expression of transforming growth factor beta (TGF‐β) and the expression and phosphorylation of Smad were also determined. Finally, endothelial cell migration was tested. POVPC did not affect the endothelial cell viability, but induce the cellular morphology transformation from cobblestone‐like endothelial cells into spindle‐like mesenchymal cells. POVPC decreased NO production and increased O2•− generation, which inhibited by NG‐nitro‐L‐arginine methylester (L‐NAME). POVPC also inhibited expression of CD31, but stimulated the expression of α‐SMA and TGF‐β as well as the phosphorylation of Smad, which partially inhibited by L‐NAME. Long term POVPC treatment promoted endothelial cell migration. These data demonstrated that POVPC could induce the changes in endothelial morphology and could uncouple eNOS activity to generate O2•− to increase oxidative stress, which may stimulate the expression and phosphorylation of TGF‐β/Smad to mediate the occurrence of EndMT. These effects may promote fibrosis remodel and spindle‐like mesenchymal cell migration to accelerate the process of atherosclerosis.Support or Funding InformationNational Natural Science Foundation of China (Grants 81370370, 81670392, 81600382 and Distinguished Young Scholar Grant 81325001), International cooperation project(2015DFA31070) and National Major Research Program (2016YFC0903000) from the Ministry of Science and Technology of China, Guangdong Natural Science Fund Committee (Grant 2015A030312009), the Changjiang Scholars Program from the Ministry of Education of China, the Guangdong Pearl River Scholars Program, the Sun Yat‐sen University Clinical Research 5010 Program.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Characterization of Microparticle Generation from Blood and Endothelial Cells during Inflammatory Stimulation
Authors: Chatterjee Victor , Meegan Jamie , Overstreet Jonathan , Coleman Danielle , Szekeres Charles , Yuan Sarah 
Published: June 2021

Microparticles (MPs) are small membrane vesicles released by activated cells in response to physical or biological stimulation. They mediate cell‐cell communication by transferring a cargo of cell surface receptors, mRNAs, and microRNAs to target cells. Recent studies have shown an altered MP production in the circulation during inflammation or sepsis. However, cell‐specific mechanisms of MP generation and their functional impact remain poorly understood. In this study, we measured MP generation from platelets, red blood cells (RBC) and leucocytes in human whole blood, as well as in cultured human umbilical vein endothelial cells (HUVEC), during stimulation by different inflammatory mediators, including tumor necrosis factor alpha (TNFα), interferon gamma (INF γ), granulocyte‐macrophage colony stimulating factor (GM‐CSF), bacterial lipopolysaccharide (LPS), thrombin, complement C5a and nitric oxide (via the NO donors DEA NONOate and SNAP). MPs isolated from blood or cell culture conditioned media were analyzed by flow cytometry using cell specific markers and Annexin V, a marker for phosphaditylserine (PS) abundant in MPs. Size‐based gating of MP was performed using a mixture of standardized beads of sub‐micron size and absolute MP numbers calculated based on reference beads of known concentration. The results showed that platelet‐derived MPs were significantly increased by C5a, thrombin, cytokines and LPS. Neutrophil‐derived MPs were increased by LPS, cytokines, GM‐CSF, C5a, and NO donors. RBC‐derived MPs were mainly increased with NO donors. In cultured endothelial cells, TNFα caused a significantly increased, whereas NO decreased production of MPs that were stained positive for ICAM‐1, VCAM‐1, PECAM‐1 and E‐selectin. Interestingly, endothelial cells were able to internalize leukocyte‐derived MPs, evidenced by confocal microscopic confirmation of their intracellular location. Furthermore, neutrophils responded to endothelial‐derived MPs by forming neutrophil extracellular traps. These findings suggest a cell‐specific production and crosstalk of microparticles with blood cell and endothelium origins during stimulation with different inflammatory mediators.Support or Funding InformationNIH HL070752, HL126646This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Evaluation of the Impact of the Traditional Medicine Formulation Goreisan and Its Components on Mesenteric Lymphatic Vessel Contraction and Lymphatic Endothelial Barrier Function
Authors: Jo Michiko , Trujillo Andrea N. , Shibahara Naotoshi , Breslin Jerome W. 
Published: June 2021

Kampo medicines are traditional herbal formulas used to treat and prevent various diseases. The Kampo medicine Goreisan is a known diuretic and is currently being tested in the clinic to reduce symptoms of surgically induced lymphedema in OB/GYN patients in Japan. The present study was aimed to clarify how Goreisan and its five different components affect lymphatic pump function. Mesenteric collecting lymphatics were isolated from anesthetized male Sprague Dawley rats, and mounted on resistance‐matched glass micropipettes in a 37 °C physiological salt solution bath for studies. The luminal pressure was set at 2 cm H2O and vessels that established intrinsic phasic contractions were studied. Contraction frequency (CF), end diastolic diameter (EDD), and end systolic diameter (ESD) were measured. From these, amplitude (AMP), ejection fraction (EF), and fractional pump flow (FPF) were calculated. Goreisan was applied at concentrations of 1–30 μg/ml. Its components, Cinnamomi Cortex, Atractilodys Rhizoma, Alismatis Rhizoma, Polyporus, and Poria were also studied at concentrations of 1–30 μg/ml. To measure barrier function, human dermal lymphatic endothelial cells (LEC) were treated with 1–100 μg/ml of each compound and transendothelial electrical resistance (TER) was determined by electric cell‐substrate impedance sensing (ECIS). The results show that treatment with Goreisan causes no significant changes in all parameters compared to baseline. However, 30ug/ml Alismatis Rhizoma caused a significant reduction in CF and FPF. The other components did not affect pumping. Goreisan also does not significant affect TER. Polyporus significantly increased TER compared to baseline, while the other components elicited no change. Collectively, the results suggest that Goreisan does not directly affect lymphatic pumping or barrier function, although at its component Alismatis Rhizoma can influence pump activity and Polyporus can enhance barrier function. By extension, in the lymphedema patients treated with Goreisan, its pharmacological action is more likely to be on renal mechanism to reduce excess body fluids. Additional work is needed to determine whether Goreisan may have direct effect to rescue impaired lymphatic vessel functions under inflammatory conditions.Support or Funding InformationThis work was supported by Rotary International Global Grants Scholarship Grant Number GG1746618.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.