bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fermented Berry Beverage Phenolics Reduced Fat Mass and Fasting Blood Glucose in High‐Fat Fed Mice
Authors: Johnson Michelle , Gonzalez de Mejia Elvira 
Published: June 2021

The objective was to determine the potential of phenolic compounds from a fermented 30:70% blueberry‐blackberry beverage to reduce obesity and hyperglycemia in three week old male diet‐induced obese C57BL/6j mice. Mice (n= 12/group) were randomized into six groups to drink: an alcohol‐free fermented beverage (AFFB) containing 65.1 ± 1.6 mg cyanidin‐3‐glucoside (C3G) equivalents/L; three doses of a phenolic extract where AFFB was eluted through Amberlite XAD‐7HP and adjusted to 6.4 (0.1x), 59.6 (1x) and 192.4 (3x) mg C3G eq./L; sitagliptin (30 mg/kg BW/d) or water as positive and negative controls, respectively. After a week, mice started on a 60% fat diet (HFD). Daily consumption of anthocyanins was 8.4, 1.1, 9.0, and 19.0 mg C3G/kg BW/d for AFFB, 0.1x, 1x, and 3x, respectively, with anthocyanins present predominantly C3G (65.8%). The BW/food intake ratio and sucrose consumption were similar (p > 0.05). Mice in the 3x group had the lowest body weight gain (10.3 ± 0.3 g) and the water group the most weight gain (16.7 ± 1.2 g, p < 0.05). The weight increase was attributed to a gain in fat mass (18.0 ± 0.8 % BW for 3x vs. 31.3 ± 1.1 % BW for water) which was correlated with visceral fat (0.8 ± 0.1 g for 3x vs. 2.4 ± 0.2 g for water), while absolute lean mass did not differ (p > 0.05). After 3 weeks of treatment, all groups had fasting blood glucose (FBG) above 126 mg/dL. At the end of the study (week 12), FBG for 1x and 3x groups was significantly lower (168 and 184 mg/dL, respectively) than the water (222 mg/dL), and the sitagliptin group (217 mg/dL, p < 0.05). Overall, phenolic compounds, mainly anthocyanins, from a fermented blueberry‐blackberry beverage reduced BW gain, fat mass accumulation, and FBG in diet‐induced obese mice.Funded by Hatch grant Division of Nutritional Sciences 20/20
Effect of Recombinant Lubricin on Coagulation Parameters in Human Blood
Authors: Fareed Jawed , Hoppensteadt Debra , Iqbal Omer , Abro Schuharazad , Sweigert Patrick , Truitt Edward , Jay Gregory , Hopkinson William 
Published: June 2021

Lubricin (proteoglycan 4; PRG4) is a proteoglycan which acts as a joint lubricant. Lubricin is present in synovial fluid and plays a major role in synovial homeostasis. A recombinant form of lubricin, rhPRG‐4 is developed to treat a variety of inflammatory diseases. (Lubris Pharmaceutical). The purpose of this study is to determine the effect of rhPRG‐4 on blood coagulation utilizing various experimental studies.rhPRG‐4 was supplemented with citrated whole blood from normal human donors (n = 10) to obtain concenration range of 0 ‐ 200 mg/mL. Activated clotting time (ACT), thrombelastographic analysis (TEG), and whole blood prothrombin time (PT), partial thromboplastin time (aPTT), and thrombin times (TT) were measured. Similarly, citrated plasma was supplemented with rhPRG‐4 at 0‐200 mg/ML to determine PT, aPTT and TT. The effect of this agent was also studied on the generation of proteases. In addition, rhPRG‐4 was also studied for its effect on platelet aggregation.rhPRG‐4 did not produce any effect on the whole blood clotting assays, such as the PT, aPTT and TT. Similarly, in the citrated plasma, none of the clotting assays were altered. rhPRG‐4 did not produce any inhibition of thrombin generation in plasma. Furthermore, it did not produce any inhibition or augmentation of agonist‐induced platelet aggregation. These studies suggest that rhPRG‐4 does not modify whole blood and plasma coagulation profile and produce and more relation of platelet function.These studies suggest that because of no potential alteration of the coagulation system, lubricin is not expected to modulate bleeding or thrombotic processes in clinical indications.
Bovine and Porcine Mucosal Heparins Exhibit Similar Biologic Profiles
Authors: Fareed Jawed , Nader Helena , Lima Marcelo , Hoppensteadt Debra , Walenga Jeanine , Jeske Walter , Kumar Emmanuel , Raake Wolfram , Bakhos Mandouh 
Published: June 2021

IntroductionIn consideration of the limited supply of porcine heparin (PH), which may result in a global shortage, the current discussions on the reintroduction of bovine heparin (BH) are timely and justified. This study compared 5 different batches each of BHs and PHs both of mucosal origin.Materials and MethodsCommercially available PHs and BH were obtained from various US and Brazilian vendors. USP reference standard heparin (Lot F01187) was used to compare the potency of these two groups of heparin utilizing ACT, aPTT, Heptest, Calcium Thrombin Time, anti‐Xa and anti‐IIa activities, as well as protamine and PF4 neutralization. The interaction with HIT antibodies was studied using a pooled HIT sera preparation. The molecular weight profile was determined by HPLC.ResultsOn a gravimetric basis, the anticoagulant activity of PH was 10‐30% higher in comparison to BH. PH exhibited higher anti‐Xa and anti‐IIa activities. The USP cross referenced potency in the anti‐Xa assay was 160+20 U/mg for PH vs 132+16 U/mg for BH. Protamine and PF4 titration profiles were comparable. In the HIT assay, there was no difference in between the two groups. .The molecular weight of BH was 17.1 ± 0.8 kDa, whereas that of PH was 16.6 ± 0.5 k Da (p>0.05).ConclusionWhile PH exhibits relatively stronger anticoagulant and anti‐protease activities, in other assays both BH and PH exhibit comparable compositional and functional profiles. Thus, BH may be considered as biosimilar to PH and can be accordingly defined and developed for clinical use.
The Allosteric Heat Shock Protein Inhibitor YM‐01 Protects Against Alcohol‐Induced Growth Arrest
Authors: Ganapathy Ekambaram , Khachatoorian Ronik , Patel Harsh , Arumugaswami Vaithilingaraja , French Samuel 
Published: June 2021

BackgroundOxidative stress is implicated in the pathogenesis of alcohol‐induced liver injury. This is, in part, mediated by the cytochrome P450 CYP2E1 which contributes to reactive oxygen species (ROS) production. YM‐01, a derivative of MKT‐077, is an allosteric heat shock protein inhibitor and modulator of the triage function of heat shock cognate protein 70 (HSC70) and may protect cells from oxidative stress. In this study, we investigated the effect of YM01 on ethanol‐induced growth arrest.MethodsHepG2 cells overexpressing CYP2E1 (E47) were pretreated with either vehicle or YM‐01 then treated with arachidonic acid (60 µM for 18 hrs), followed by iron nitrilotriacetic acid (Fe‐NAT, 25 µM for 5 hrs), and finally with ethanol (100 mM for 48 hrs). MTT and western analysis was then performed.ResultsIn the CYP2E1‐expressing ethanol‐treated cells, cell growth was arrested as determined by cell viability (MTT assay). This arrest was relieved by treatment with YM‐01. Further, western analysis showed that pretreatment with YM‐01 prevented 4‐hydroxynonenal (4‐HNE) adduct formation induced by ethanol (the most abundant lipid peroxidation end product). In addition, we determined ubiquitination levels by Western analysis to determine role of the ubiquitin‐proteasome pathway.ConclusionThis study indicates that YM‐01 decreases 4‐HNE adduct formation in CYP2E1‐expressing cells and protects against alcohol induced cell growth arrest.
GPC3‐CD81 axis in the HCV mediated liver carcinogenesis
Authors: Xue Yuhua , Bowen Bill , Orr Anne , Koral Kelly , Haynes Meagan , Bell Aaron , Paranjpe Shirish , Mars Wendy , Michalopoulos George 
Published: June 2021

Glypican‐3 (GPC3) is highly over‐expressed in human hepatocellular carcinomas (HCC). GPC3 decreases liver regeneration by interacting with CD81. Other groups have shown that activation of CD81 increases phosphorylation of Ezrin (p‐Ezrin), which inhibits Hippo pathway. Hepatitis C virus (HCV) interacts with human CD81 by E2 protein to enter into hepatocytes. We investigated the mechanism of HCV and GPC3‐CD81 axis in liver carcinogenesis. HepaRG human hepatoma cell line was treated with E2 protein to evaluate the Hippo activity. Rat hepatocytes were treated with CD81 agonist antibody to determine if CD81 related Hippo activity depends on p‐Ezrin. Partial hepatectomy (PHx) was used to evaluate the dynamic changes of GPC3‐CD81 axis with Hippo activity. Human and rat HCC cell lines were used to define GPC3‐CD81 interactions. HCC tissue array was used to assess the expression characteristic of CD81‐GPC3. After E2 treatment, Hepa RG cells had higher activity of Hippo. Treatment of rat hepatocytes with CD81 agonist antibody led to lower Hippo activity and higher p‐Ezrin. PHx shows that GPC3 increases at the early stage of liver regeneration while CD81 decreases, while Yap activity increases. Both human and rat HCC cell lines have dramatically decreased CD81 expression levels. HCC tissue array also shows absence of CD81 in most HCC.ConclusionsDependent on CD81, GPC3 may function as tumor‐suppressor gene by modulating YAP expression. Persistent E2 HCV protein interacts with CD81 to decrease Yap via Hippo. Hepatocytes with no expression of CD81 would have a growth advantage to undergo clonal expansion, facilitating formation of HCC. After absence of CD81, GPC3 may be unable to control the Hippo, and this maybe the reason for its high expression in HCC.
Regulation of Actin Interacting Protein Drebrin by Mutations in HSPB1
Authors: Mata Marina , Sun Xiankui , Fink David 
Published: June 2021

Heat shock protein B1 (HspB1) is an ATP‐independent molecular chaperone with a conserved α‐crystalline domain in the C‐termini region. A number of mutations in HspB1 have been identified in families with Charcot Marie Tooth type IIF (CMT‐IIF) disease or with distal hereditary motor neuropathy II (dHMN II). Mutations outside the crystalline domain (P39L, G84L, P128S, P182L) associate with dHMN. Mutations S135F, R136W and R127W associate with a CMT‐IIF phenotype.Wild type (wt) HspB1 reduces RhoA activation by translational inhibition of PDZ‐RhoGEF, a RhoA specific GEF. Decreased expression of PDZ‐RhoGEF by HspB1 was accomplished by enhancing expression of specific microRNAs (miR20a and miR128) that inhibit the translation of PDZ‐RhoGEF and resulted in an increased neurite growth response. The regulatory effect of HspB1 on RhoA activity is blocked by mutation in S135F of HspB1 but not by R136W and R127W. This suggested either that these HspB1 mutations alter independent cellular mechanisms or that their effects may converge downstream of RhoA GTPase signaling to cause similar disease phenotype.We constructed lentiviral vectors carrying full length human HspB1 cDNA of wt or mutant S135F, R136W or R127W. Cortical neurons derived from SD rat pups E15 were transfected to expressed wt or mutant HspB1transgene to determine their effect on actin cytoskeleton.The studies showed S135F, R16W and R127W mutations cause HspB1 to partition more readily with the cytoskeletal fraction and enhance the binding of HspB1 to F‐actin leading to the loss of Drebrin, an F‐actin side‐binding protein, from the F‐actin domain. These findings highlight the diverse mechanisms by which HspB1 functions as an important regulatory protein of the actin cytoskeleton.
Use of Daniovision™ to Measure Activity in 3rd Instar Drosophila melanogaster Larvae
Authors: Graham Stephanie , Rogers Ryan , Alper Richard 
Published: June 2021

The noncompetitive GABAA receptor antagonist picrotoxin (PTX) decreases activity in 3rd instar Drosophila larvae (Eur J Neurosci, 2006). The purpose of these studies was to replicate the data obtained by Stilwell et al. using an automated data acquisition/data reduction system, Daniovision™ by Noldus.Larvae were extracted from food (Carolina Biologic Supplies 4‐24) in 20% sucrose at the 3rd instar stage. The larvae were randomly placed into vials containing vehicle (20% sucrose + 0.3% DMSO) or PTX from 0.03‐3 mM: n = 20‐30 larvae/treatment. After incubation for 2‐24 hr, individual larvae were placed in separate wells of a 6‐well culture plate previously coated with 2% agar, then placed in the Daniovision™ apparatus. The activity of each larva was digitized and recorded for 5 minutes using Ethovision software.PTX reduces 3rd instar larvae activity in a concentration‐dependent manner, consistent with findings reported by Stilwell et al. Incubation in 1 mM PTX for 4 or 24 hours decreased activity, with consistent reduction produced by 3 mM, variably by 0.3 mM, and never by 0.03 mM. PTX, 0.03‐3 mM, did not alter activity in a mutant strain, Rdl, known to have a point mutation in the PTX binding site on the GABAA receptor conferring a lower affinity for PTX.We have shown that an automated system can be used to quantify activity in 3rd instar Drosophila melanogaster larvae. As human observation and review of videos is not required, and multi‐well plates can be used, we have refined an in vivo high‐throughput screen for anti‐seizure compounds.Supported by funds from the University of Saint Joseph.
β‐caryophyllene‐complexed in β‐cyclodextrin Produces Antihyperalgesic Activity in Animal Model for Fibromyalgia
Authors: Quintans Jullyana , Quintans Júnior Lucindo , Araújo Adriano , Barreto Rosana , Sluka Kathleen 
Published: June 2021

β‐caryophyllene is a dietary cannabinoid and it is a major plant volatile found in large amounts in the essential oils of many different spice and food plants, such as oregano (Origanum vulgare L.), cinnamon (Cinna‐momum spp.) and black pepper (Piper nigrum L.). The analgesic activity of β‐caryophyllene‐complexed in β‐cyclodextrin (CD‐CAR) was assessed in an animal model of fibromyagia (chronic non‐inflammatory muscle pain). Male Swiss mice were pretreated with CD‐CAR (10 or 20 mg/kg, p.o.) or vehicle (neutral saline). They were screened for mechanical hyperalgesia of the paw and muscle (von Frey and tweezer), motor coordination (Rota Rod) and muscle strength (Grip Strength Meter) beginning 60 min after treatment and continuing through 15 daysCD‐CAR, all doses, induced a significant reduction (p<0.05, p<0.01 or p<0.001) of mechanical hyperalgesia of the paw nd muscle in the chronic non‐inflammatory muscle pain model that lasted for 15 days. CD‐CAR produced no significant change in motor coordination or muscle strength. Therefore, we show that CD‐CAR reduces pain behaviors in an animal model for fibromyagia.Financial Support: CNPq/Brazil, FAPITEC/Brazil and FINEP/Brazil
Effects of Incisional Pain on Food‐Maintained Responding under Progressive Ratio Schedules of Reinforcement
Authors: Warner Emily , Krivitsky Rebecca , Atherton Phillip , Pitre Travis , Cone Katherine , Lanpher Janell , Giuvelis Denise , Bilsky Edward , Stevenson Glenn 
Published: June 2021

The current set of experiments characterized the effects of incisional pain on food‐maintained responding under progressive ratio operant schedules. Rats were trained to press a lever for food on a FR 1 schedule of reinforcement. The ratio requirement was gradually increased to 5, and following stable responding rats were assigned to one of four progressive ratio schedules: PR2, PR5, Log1 (j=0.17), Log2 (j=0.20). The effects of reinforcer type (grain vs. sugar pellets) limited hold duration (min), and ratio progression (logarithmic vs. arithmetic) were manipulated to develop a schedule with (1) low intra‐animal variability in breakpoint, (2) high rates of responding, and (3) session lengths shorter than 60 min to facilitate future drug testing. The Log2‐LH15 min schedule was chosen as terminal schedule. Rats received either incision + anesthesia or anesthesia only. Parallel studies assessed mechanical allodynia. For rats responding for grain, the incisional pain manipulation produced significant reductions in breakpoints and mechanical thresholds across three days. In contrast, for rats responding for sugar, incisional pain produced no change in breakpoints and decreased mechanical threshold only at post‐6hr. Given that PR schedules are interpreted to reflect workload and/or motivation, these schedules may have utility as a preclinical model for assessing pain / motivation / cognition interactions in rodents.Funding: NIAMS/NIH 2R15AR054975‐02A1 to GWS
Interacting microRNA and mRNA genes identify potential therapeutic targets for NSCLC
Authors: Black Esther , Krentz Madeline , Wu Wenhui , Stromberg Arnold 
Published: June 2021

Lung cancer patients die from drug‐resistant, metastatic disease. Lung cancers have high mutation rates. We propose that analysis of gene expression data can uncover new drug targets and strategies to improve outcomes in Non‐Small Cell Lung Cancer (NSCLC). Partial Least Squares (PLS) regression analysis coupled with bioinformatics processing can identify microRNA(miR) and mRNA interactions with druggable potential.miR/mRNA signatures that predict NSCLC response to epidermal growth factor (EGFR) inhibitors were assembled using SAS. mRNA were regressed on miR genes for 8 samples. mRNA probesets that interacted at least 5 miR genes moved forward to biological validation by gene silencing. STRING used probeset/gene IDs to map protein‐protein interactions (PPIs). From the PPIs, the interactions of genes within two edges determined whether targets interact with the EGFR network.After cross‐validation, 8 interactions were retained, and one physical interaction was confirmed. EIF4B, RAB8A, and PPP1CB interact in the PI3K/AKT cascade while KLK8 and PRKX interact with TGFß signaling; both pathways intersect EGFR. We silenced EIF4B, RAB8a, and PPP1CB +/‐ EGFR inhibition in NSCLC cells. Silencing of EIF4B and RAB8A did not induce cell cycle arrest or death. PPP1CB is a potential novel target. PLS modeling and bioinformatics was used to build a logical and repeatable workflow with gene expression data to identify new drug targets/strategies. Of 14000 possible interactions, 8 genes were processed, and 3 were chosen for biological validation. PLS modeling and bioinformatics analyses will be optimized.KY Bioinformatics Research Infrastructure Network Pilot Project (NIH‐8 P20 GM103436‐12)
Inhibition of Micro‐RNA 146a Expression in Lung Fibroblasts by Fluticasone Propionate, Salmeterol Xinafoate, and Related Agents
Authors: Toews Myron , Schulte Nancy , Bang Jarrod , Rennard Stephen , Liu Xiangde 
Published: June 2021

Micro‐RNA (miR) 146a is induced by inflammatory stimuli, and its pro‐inflammatory targets suggest that it may limit the inflammatory response. Fluticasone propionate (FP) and salmeterol xinafoate (SX) are used together to treat asthma, an inflammatory disease. Their effects on interleukin 1‐beta (IL‐1β)‐induced expression of miR‐146a by HFL‐1 lung fibroblasts were evaluated by quantitative RT‐PCR. Exposure of cells to the anti‐inflammatory steroid FP (1 nM) for 24 hr caused a nearly 70% inhibition. Two beta‐2 adrenergic receptor agonists, the long‐acting SX (10 nM) and the short‐acting albuterol (Alb, 10 μM) also inhibited miR‐146a expression, but only by 25‐35%. Prostaglandin E2 (100 nM) and the direct adenylyl cyclase activator forskolin (Fsk, 30 μM) both inhibited miR‐146a expression similarly to SX and Alb, suggesting cAMP elevation as the mechanism for these agents. Two primary isolates of human lung fibroblasts showed similar decreases in miR‐146a as for HFL‐1 cells when treated with FP, SX, or Fsk. FP has anti‐inflammatory effects in the airways that are augmented by concurrent SX, yet both FP and SX reduced fibroblast expression of anti‐inflammatory miR‐146a. Whether this is a direct effect of these agents or due to their overall inhibition of inflammation remains to be defined. Reduction in miR‐146a levels by these agents might limit their anti‐inflammatory actions, perhaps to prevent excess inhibition of inflammatory pathways.Supported by Glaxo Smith Kline Respiratory Division Investigator‐Initiated Grant 200164.
Involvement of Two Distinct Signalling Pathways in IGF‐1‐Mediated Central Control of Hypotensive Effects in Normotensive and Hypertensive Rats
Authors: Tseng Ching‐Jiunn , Cheng Pei‐Wen 
Published: June 2021

Insulin‐like growth factor‐1 (IGF‐1) is abundantly expressed in the nucleus tractus solitarii (NTS). In a previous study, we revealed that the induction of nitric oxide (NO) production in the NTS reduces blood pressure (BP). It is well known that both acute and chronic administration of IGF‐I reduce BP. The aim of this study was to evaluate the short‐term hypotensive effect of IGF‐1 in the NTS and to delineate the underlying molecular mechanisms of IGF‐1 in the NTS of normotensive WKY rats and spontaneously hypertensive rats (SHRs).Microinjections of the phosphatidylinositol 3‐kinase (PI3K) inhibitor LY294002 and the MAP kinase‐ERK kinase (MEK) inhibitor PD98059 into the NTS in WKY rats and SHRs were used to study the involvement of IGF‐1‐induced depressor effects.An IGF‐1 (7.7 pmol) injection into the NTS resulted in a significant decrease in BP and HR in WKY rats and SHRs. Immunoblotting and immunohistochemical analysis showed that the microinjection of LY294002 (0.6 pmol) or PD98059 (3.0 pmol) into the NTS attenuated the IGF‐1‐induced depressor effects and Akt or ERK phosphorylation in WKY rats. An attenuation effect of LY294002, but not PD98059, was found in the SHRs.These results suggest that distinct Akt and ERK signalling pathways mediated the IGF‐1 control of the central depressor effects in WKY rats and SHRs. ERK signalling defects may be associated with the development of hypertension.
Intralipid Reverses Propofol‐Mediated Hypotension in Rats
Authors: Bourque Stephane , Panahi Sareh , Mittal Rohan , Moher Conrad , Gragasin Ferrante 
Published: June 2021

IntroductionPropofol is a commonly used anesthetic. Despite its favourable safety profile, propofol causes hypotension which can result in end‐organ hypoperfusion. Intralipid is a lipid emulsion that has been shown to reverse the vasodilatory effects of propofol in isolated vessels; however, whether these effects are recapitulated in vivo is not known. The objectives of this study were to determine if intralipid reverses the hypotensive and anesthetic effects of propofol in rats.MethodsUnder isoflurane anesthesia, male Sprague Dawley rats were instrumented with indwelling catheters for mean arterial pressure (MAP) assessments as well as subdural electrodes for cortical activity assessments by electroencephalography (EEG).ResultsPropofol (10 mg/kg IV) caused hypotension (55±2% drop in MAP, P<0.001) and intralipid (4mL/kg IV) caused greater reversal (80±9%) of blood pressures compared to saline (19±1%; P<0.001). Blockade of the autonomic nervous system with chlorisondamine (2.5 mg/kg IV) caused marked hypotension (56±3% lowering of MAP, P<0.001) which could be reversed with a constant infusion of phenylephrine (300 μg/kg/hr); under these conditions, propofol nevertheless caused hypotension (12±4% lowering of MAP) which was completely reversed by intralipid. Propofol‐induced cortical burst suppression was not affected by intralipid (2±3%), saline (‐4%) or 20% BSA (‐2±1%; P=0.27).ConclusionThese results demonstrate that intralipid reverses propofol‐mediated hypotension with minimal effects on its anesthetic profile. Intralipid could be particularly useful as a rescue against propofol in patients prone to hemodynamic instability such as the elderly.This work was funded by a grant from the University Hospital Foundation.