bluuebunny/crossref_metadata_2025 huggingface dataset:
Grip strength and metabolomic effects in mice fed a high‐nitrate beet diet (647.32)
Authors: Kemp Michael , Dever Joseph , Thompson Amber , Scholl Chris , Barnes David
Published: June 2021

Beets (Beta vulgaris) are large edible tap roots vegetables belonging to the chenopod family. In addition to being a good source of essential micronutrients and minerals, beets also contain high levels of nitrates. Diets high in beet and nitrates have been demonstrated to improve exercise capacity, muscle contractility, and blood pressure. While these results have been replicated several times little is known about the mechanism of action. In this study we investigated grip strength and metabolomic changes in response to diets containing sodium nitrate, and beets providing nitrates at 5.5, 2.75, 1.37 mmol/g BW/day. Grip strength studies demonstrated a dose response improvement in the mice eating the beet diet, with a statistically significant peak grip strength improvement of 25%. These results are consistent with previous reports that beet juice improves muscle contractility. Untargeted metabolomic analysis in two separate studies indicated an upward trend (p=0.06) of plasma sphingosine 1‐phospate (S1P) concentrations in mice fed a beet diet (5.5mmol/gBW/day) compared with mice fed a control diet. Increased S1P has been previously correlated with exercise performance, muscle repair and activation of muscle stem cells. Overall these results suggest that high‐nitrate foods such as beets may improve muscle contractility through a mechanism of action which may include S1P and targets under its influence.
Reserpine‐induced gastric ulcer preventive activities of Malva verticillata L. aqueous extract in Kuming mice (647.6)
Authors: Zhao Xin , Li Guijie , Song Jia‐Le , Suo Huayi , Wang Qiang , Qian Yu
Published: June 2021

Malva verticillata L. (MVL) was purchased for the evaluation of gastric ulcer preventive effect on Kunming mice. We used an animal model to check gastric ulcer preventive activities of MVL in vivo. The mice were divided into five groups (normal, control, 250 mg/kg and 500 mg/kg MVL dose and ranitidine groups). The mice in MVL groups was treated with MVL aqueous extract for 4 weeks, and the last day the gastric ulcer was induced by reserpine in control, MVL and ranitidine groups. 24 h later after dissection, we tested the area of gastric ulcer, serum levels of motillin (MOT), somatostatin (SS), substance P (SP) and vasoactive intestinal peptide (VIP), serum proinflammatory cytokines of interleukin 6 (IL‐6), interleukin 6 (IL‐12), tumor necrosis factor‐α (TNF‐α) and interferon‐γ (IFN‐γ), gastric juice volume, gastric juice acidity and iNOS, COX‐2 gene expressions. The gastric ulcer area was significantly down‐regulated by MVL increased the SS and VIP, decreased the MOT and SP serum levels. Malva verticillata L. reduced the levels of serum proinflammatory cytokines of IL‐6, IL‐12, TNF‐α and IFN‐γ compared to control group. MVL also decreased the expression of iNOS and COX‐2. And the 500 mg/kg of MVL showed the more visible trends. These results suggest that MVL showed a good gastric ulcer preventive effect.
Cardiac compensation for lack of trabeculae in ErbB2‐deficient zebrafish (648.3)
Authors: Fleming Nicole , Samsa Leigh Ann , Liu Jiandong
Published: June 2021

Congenital heart disease (CHD) is one of the major causes of birth defects among infants in the United States. CHD can be characterized by malformations during heart development. Initially, the heart is a smooth, linear tube of cardiomyocytes lined with specialized endothelial cells. Following cardiac looping, the heart chambers undergo dramatic changes in chamber topology, most notably the formation of luminal, muscular protrusions called trabeculae. While altered trabeculation leads to severe cardiac defects, complete failure to form trabeculae results in embryonic lethality in human, mouse, chick, and zebrafish due to cardiovascular failure. We hypothesize that cardiac trabeculation is critical for cardiac function in the developing heart. Due to their external fertilization and genetic tractability, we use the zebrafish embryo to observe trabeculation. Zebrafish cardiac trabeculation requires both genetic and non‐genetic mediators: requiring input from the Neuregulin/ErbB2 signaling pathway and hemodynamic forces, respectively. To understand the cellular mechanism of progressive cardiac dysfunction in the absence of trabeculae, we inhibited ErbB2 signaling in the zebrafish model. Preliminary data indicates increased compact myocardial thickness (2.09 ± 0.19 μm in DMSO treated fish vs. 3.96 ± 0.35 μm in ErbB2‐inhibitor treated fish; p <0.05) and sarcomere length (1.30 ± 0.15 μm in DMSO treated fish vs. 1.92 ± 0.18 μm in ErbB2‐inhibitor treated fish; p <0.001), a phenotype reminiscent of pressure overload‐stimulated cardiac hypertrophy. This compensatory mechanism suggests that trabeculae may play a role in optimizing chamber topology for efficient cardiac function.

Grant Funding Source: Supported by: Initiative for Maximizing Student Diversity (IMSD): 5R25GM055336 and BBSP
Clinical evaluation of heart failure: agreement among tests (648.4)
Authors: Pandey Amit , Penny William , Bhargava Valmik , Lai N. Chin , Hammond H. Kirk
Published: June 2021

Congestive heart failure (CHF) is the most common cause of nonelective hospitalization of subjects over the age of 65 in the US. CHF clinical status is evaluated by ejection fraction (EF), symptoms, and exercise tolerance. Although difficult to apply clinically, a gold standard of left ventricular (LV) function is the rate of LV pressure development (+dP/dt). We tested the hypothesis that EF, symptoms, and exercise treadmill test (ETT) duration would correlate with each other and with +dP/dt. Subjects with symptomatic CHF (62 ± 2 yo; EF 30 ± 2%; n = 27) underwent measurement of EF, symptoms (Kansas City Cardiomyopathy Questionnaire Score, KCCQS), ETT duration, and +dP/dt (via cardiac catheterization). EF and +dP/dt were measured before and during dobutamine infusion. Results were compared using Pearson’s correlation analysis. Overall, tests correlated poorly. The only significant correlations were between KCCQS and +dP/dt. KCCQS correlated with both basal +dP/dt (r = ‐0.47, p = 0.021) and stimulated +dP/dt (r = ‐0.63, p < 0.001). KCCQS and stimulated +dP/dt were correlated regardless of ischemic (r = ‐0.69, p = 0.026) or non‐ischemic etiology (r = ‐0.56, p = 0.038).

Conclusion: EF, ETT, and symptoms are not correlated in CHF patients. Using +dP/dt as a gold standard, EF and ETT do not correlate as well with LV function as symptoms do. EF and ETT may be less effective measures of LV function than commonly believed.

Grant Funding Source: Supported by National Institutes of Health grants (P01 HL66941, HL088426, HL081741, and HL107200)
Gene network analysis of liver allografts preserved with machine perfusion (649.10)
Authors: Paranjpe Shirish , Fontes Paulo , Vodovotz Yoram , Michalopoulos George
Published: June 2021

Machine perfusion (MP) with a cell‐free oxygen carrier solution (CFOCS) was compared to cold static preservation (CSP) in a porcine model. Methods: Two groups of 6 animals underwent liver transplantation after 9 hours of cold ischemia time. Clinical and laboratorial data was obtained in additional to serial histological and electron microscopy analysis. Affymetrix gene‐arrays were performed on liver tissues from both groups. A cytokine profile was obtained from liver tissues and perfusate with Luminex. Additional metabolomics analysis (Metabolon) was performed. Results: The MP group had 100% survival and the CSP group had 33% survival. A significant (p<0.004) benefit of MP was detected after 16 different variables were analyzed. Over‐expression of genes associated with general metabolic functions, anti‐inflammatory, regenerative and protective mechanisms against free radicals were observed in the MP group. Genes associated with hepatic system development and cellular proliferation were significantly up‐regulated in the MP group. Genes associated with critical metabolic networks were significantly down regulated in the CSP group compared to MP livers. The lipid metabolic pathway was up regulated in the CSP group. Signaling pathways that are critical for liver growth (NF‐kB, PI3K/AT, ErbB) were significantly up‐regulated in MP liver samples when compared to CSP group. INF‐α, IL‐1β, IL‐1RA, IL‐4, IL‐10, IL‐12p40, IL‐18 and TNF‐α showed significant differences between the groups in liver tissues analyzed during preservation. Ingenuity® pathway analysis of functional groups revealed that genes associated with liver damage were significantly down regulated in the MP group. The CSP group showed significant up‐regulation of genes associated with liver pathology when compared to the MP group. Conclusions: MP with a CFOCS up‐regulated a regenerative and prosurvival response when compared to CSP.

Grant Funding Source: NIH GrantNo.CA35373,CA103958 & Rangos Fund for Enhancement of Researchin Pathology.
T3 receptor‐beta agonists GC‐1 (Sobetirome) and KB2115 (Eprotirome) stimulate hepatocyte proliferation in mice (649.5)
Authors: Feliciano Tamara , Monga Satdarshan
Published: June 2021

T3 hormone is known to be a strong inducer of hepatocyte proliferation in rats and mice. Recent work has shown that the molecular mechanism for this may be Protein Kinase A‐dependent activation of the β‐Catenin signaling pathway. This finding has shed light on the potential therapeutic use of thyroid hormone in select cases of liver failure. However, systemic side effects of T3 may preclude its clinical use. GC‐1 and KB2115 are T3 hormone analogs that bind selectively to the T3 hormone β‐receptor and do not seem to produce the undesirable cardiovascular effects. We investigate whether GC‐1 and KB2115 are able to induce proliferation in mouse hepatocytes. Nine C57/BL6 2‐month old male mice received eight daily intraperitoneal injections with GC‐1, KB2115, or DMSO (control) at 0.3mg/Kg body weight. Livers from mice injected with GC‐1 and KB2115 showed increased levels of Cyclin D‐1 on Western Blot compared with controls. We also saw an increase in Beta‐Catenin, Cyclin D‐1, and Ki‐67 by immunohistochemistry, without significant difference in serum biochemistry. This suggests that selective thyromimetics are likely activating β‐Catenin signaling pathway in a similar manner to T3 hormone, and with further study may prove useful as an adjunct therapeutic option in select cases of liver failure.

Grant Funding Source: 1RO1DK62277 to SPM
Necrotizing enterocolitis leads to increased intestinal permeability mediated through differential expression of tight junction proteins (650.7)
Authors: Ravisankar Srikanth Sankar , Tatum Rodney , Garg Parvesh , Shekhawat Prem , Chen Yan‐Hua
Published: June 2021

Necrotizing enterocolitis (NEC) is a devastating disease with a prevalence of 7% in premature neonates weighing <1500 grams and an estimated death rate of 20‐30%. The precise etiology remains unknown and it is believed to be caused by a disturbance in the delicate balance of multiple pathophysiological factors. In this study, 4‐day old C57BL/6 mice pups were fed 50 µl of 33% Esbilac formula every 3 hours, asphyxiated twice daily (100% N2 for 60 sec) followed by cold stress at 4°C for 10 min to induce NEC and study the tight junction (TJ) proteins. Dam‐fed pups from the same litter served as controls. Histopathology results revealed successful induction of NEC up to grade 2, where sloughing of the epithelial cells at the tips of the villi with mild villous necrosis was seen. Western blot showed decreased expression of claudin‐1 and an increased expression of claudin‐7 in the NEC intestines compared to control pups. The TJ permeability assay was performed by injecting biotin tracer into the lumen of the small intestine of both control and NEC pups. Biotin tracer diffused easily through the intercellular space of intestinal epithelial cells in NEC mice, suggesting loss of integrity of the TJ barrier function. We conclude that there is disruption of TJs mediated through differential regulation of TJ proteins in our NEC mouse model.

Grant Funding Source: NIH
Matching the mechanical properties of the brain: histological performance of soft elastomeric wires designed for use in neural interface devices (651.15)
Authors: Kolarcik Christi , Hanner Jenna , Snyder Noah , Du Zhanhong , Sapp Shawn , Luebben Silvia , Cui X.
Published: June 2021

A number of factors contribute to the failure of microelectrodes implanted into the central nervous system and designed for therapeutic purposes. We hypothesized that more closely matching the mechanical properties of the microelectrode to those of the brain in combination with biomolecule coatings would promote a lower inflammatory response and less scar tissue encapsulation. In vitro cell culture studies were used to evaluate novel elastomeric wires with Young’s modulus lower than 1 MPa; the L1 protein coating was compared to laminin‐coated and uncoated microelectrodes. In vivo work included characterization of the tissue response associated with microelectrode implantation into the subthalamic nucleus of rats (n = 4/group) at acute and chronic time points. Commercial tungsten microelectrodes with Young’s modulus of 411,000 MPa were used as a standard (stiff control). The in vitro results indicate that soft wires had greater neuronal but less astrocyte and microglial attachment than stiff wires. Additional benefits were noted with the L1 protein coating. In vivo histological assessment indicates the superior performance of soft wires with the L1 protein coating: increased neuronal density (NF200 staining), decreased microglial presence (Iba1 staining), decreased astrocyte activation (GFAP staining) and decreased neuronal cell death (NeuN/Caspase‐3 colocalization) when compared to control stiff wires either with or without L1 coating and to soft wires without L1. This work contributes to evidence supporting the use of soft microelectrodes and the L1 protein coating.

Grant Funding Source: Supported by DARPA Contract W31P4Q‐11‐C‐0134.
Double‐edged effect of early reperfusion on arrhythmias and death after transient coronary artery occlusion in the rat (652.16)
Authors: Liao Kate , Hong Daphne , Chen Kuen‐Bao , Kuo Eva , Lai Ted
Published: June 2021

Reperfusion of blood supply following a sustained period of ischemia can be injurious to the heart. Here, we monitored cardiac reperfusion in rats subjected to 10 min of wide‐area coronary artery occlusion (CAO) by injecting them with the Evans blue dye, a visible inert tracer. The rats were then divided into three groups depending on their rate of reperfusion. Group 1 consisted of rats that showed no signs of early reperfusion within the 10 min period of ischemia, and these rats demonstrated a moderate rate of sudden cardiac death within the ischemic period. Group 2 included rats that showed early signs of reperfusion, and these rats demonstrated a higher rate of cardiac death than those in Group 1. Rats in Group 3 had poor ligation such that they sustained partial reperfusion almost immediately post‐CAO, yet surprisingly, this group demonstrated highest rate of cardiac death during the ischemic period. In contrast to mortality during the ischemic period, rats that survived the first 10 min of ischemia then received ligation‐removal and thus full‐blown reperfusion. Interestingly, surviving rats from Group 3 now has best chance of continued survival post‐reperfusion, whereas rats from Group 2 had poor survival rate and no rats from Group 1 survived reperfusion. To better understand the pre‐conditioning effect of early reperfusion on full‐reperfusion‐mediated sudden cardiac death, we challenged rats to 5 min of small‐area CAO. This model produced 100% survival rate during the ischemic period, and therefore all rats survived to receive full‐reperfusion. In this model, rats from Group 1 demonstrated highest rate of sudden cardiac death after full‐reperfusion, whereas rats from Group 2 demonstrated higher rate of survival and all rats from Group 3 survived reperfusion. Taken together, early reperfusion of blood supply may induce sudden death of the ischemic heart, but if the heart survived this earlier attack, it becomes more resistant to full‐reperfusion‐mediated sudden cardiac death.

Grant Funding Source: Supported by the National Science Council of Taiwan
Anti‐fibrillatory effects of F17727, a new IKur blocker, in rats and rabbits with myocardial infarction or heart failure (652.17)
Authors: Junquero Didier , Latil Alain , Letienne Robert , Pignier Christophe , Verscheure Yvan , Mougenot Nathalie , Le Grand Bruno , Hatem Stéphane
Published: June 2021

Atrial fibrillation (AF) can be triggered by myocardial infarction (MI) or heart failure (HF). The blockade of the ultra‐rapid outward rectifier potassium current IKur is an attractive goal to control the repolarization of cardiac atria. F17727 was evaluated in patch‐clamp assays, in rat post‐MI and in HF in rabbit and dog. F17727 blocked human IKur (IC50=1.8µM) and did not impact IKr, IKs, INa. In rats 2 months after coronary artery ligation, the left atria surface increased by 50‐150%; transœsophagial electrical bursts induced AF which lasted 5‐21s in vehicle‐MI rats but lower than 5s in sham. An acute iv treatment by F17727 (2.5 and 5mg/kg) significantly reduced AF induction at the level of sham. In rabbits, 2‐week left atria tachypacing up to 1200bpm triggered electrical and structural remodeling; electrical stimuli induced AF. An acute iv treatment by F17727 (racemate, 2.5mg/kg) reduced AF (mean spontaneous and induced AF duration of 4±4s and 82±49s) when compared to vehicle (165±75s and 261±45s respectively). In dogs with HF, F17727 did not reduce electrical stimuli‐induced AF and the role of IKur in atrial repolarization is questionable. Hence the selective IKur blocker F17727 prevents the occurrence of AF induction in rats and rabbits with different AF substrates during MI and HF. The F17727 pharmacological profile reinforces the interest of IKur blockade as a rhythm control strategy for AF treatment in humans.
Mortality in patients in providing emergency medical care at a pre‐hospital stage (652.18)
Authors: Dubitsky A. , Karp L.
Published: June 2021

Introduction. Providing of emergency medical care to the patients in the pre‐hospital phase has a great importance in medicine.

Material. The analysis was made of the calls of Astana city ambulance teams to patients who have been pronounced dead in the presence of doctors from 2006 to 2012. In total, 1002 cases were registered.

Results. By nosology the patients were distributed as follows: acute coronary pathology (69.7%), strokes (12.7%), pulmonary arterial thromboembolism (6.0%), asthma status (2.8%), infectious‐toxic shock (2.2%) and other diseases (6.6%). In 137 cases, the pain was assessed as “very severe”, in 88 ‐ as “severe”. Acute cardiovascular insufficiency of 2nd and 3rd degree according to Killip was observed in 132 patients.115 patients had baseline sinus tachycardia, other arrhythmias were in 19 patients.

60 patients were observed with a violation of A‐V conduction (1st degree A‐V block ‐ 39 cases, 2nd degree A‐V block of Mobits II type ‐ 4 cases and 3rd degree A‐V block ‐ 17 cases). Myocardial infarction was localized in anterior wall in 141 patient and in 87 patients ‐ in the rear wall. Right ventricular myocardial infarction was in 3 patients. Pathologic Q wave on the electrocardiograms was formed in 139 patients.The rise of ST interval above contour line for more than 5 mm was observed in 186 patients.16 patients were detected with the prolongation of QT interval above normal, calculated by Bazett formula.
Enhanced oxidative stress/DAPK3/Akt/ERK signaling accounts for estrogen exacerbation of cardiac dysfunction caused by ethanol in male rats (652.20)
Authors: El‐Mas Mahmoud , Abdel‐Rahman Abdel
Published: June 2021

We have previously shown that acute ethanol reduces blood pressure (BP) and cardiac output in female, but not in male, rats. Because these effects of ethanol were estrogen (E2)‐dependent, we investigated whether E2 treatment of male rats uncovers a hypotensive effect for ethanol, and whether left ventricular (LV) dysfunction, due to oxidative damage, contributes to the ethanol‐E2 hemodynamic interaction. The effect of ethanol (0.5, 1 or 1.5 g/kg; i.v.) on BP, LV function, and oxidative stress markers in male rats treated with E2 (1 μg/kg i.v.) or the vehicle were investigated. While ethanol had no effect on BP in vehicle‐treated rats, it caused dose‐related falls in BP in E2‐pretreated rats. This was associated with (i) compromised myocardial function as suggested by the reductions in LV developed pressure (LVDP), end diastolic pressure (LVEDP), and rate of rise in LV pressure (LV dP/dtmax), and (ii) oxidative damage as evidenced by the increases in LV generation of reactive oxygen species, protein expression of the malondialdehyde adducts, and phosphorylated death‐associated protein kinase‐3 (DAPK3), Akt, and ERK1/2. Together, E2 propagates LV dysfunction and hypotension in response to ethanol in male rats via facilitation of the myocardial oxidative stress/lipid peroxidation/DAPK3/Akt/ERK1/2 cascade. Because E2 was administered acutely, its effects are likely to involve rapid nongenomic mechanisms.

Grant Funding Source: Supported by NIAAA [2R01 AA014441‐07].
Novel roles of chromogranin A and its peptide, catestatin, in cardioprotection (652.3)
Authors: Schilling Jan , Avolio Ennio , Zemljic‐Harpf Alice , Headrick John , Fridolfsson Heidi , Kassan Adam , Bandyopadhyay Gautam , Siddiqui Jawed , O'Connor Daniel , Roth David , Patel Hemal , Mahata Sushil
Published: June 2021

Chromogranin A (Chga) is abundant in (neuro) endocrine cells, and its derived peptide catestatin (CST) acts as an antihypertensive, cardioprotective, pro‐angiogenic, and insulin‐sensitizing peptide. The role of Chga and CST in ischemic preconditioning (IPC), an endogenous cardioprotective response, is unknown. We hypothesized that CST in the setting of diet‐induced insulin‐resistance would modulate the response of the heart to IPC. We used wild‐type (WT), Chga knockout (KO) and CST KO mice. Animals were fed normal chow (NCD) and 60% high fat diet (HFD for 3 months). IPC‐induced cardioprotection was abolished in both NCD and HFD fed CST‐KO mice, implicating a novel role for CST in IPC. Decreased phosphorylation of Akt and GSK‐3β in CST‐KO mice suggests involvement of the RISK pathway. HFD diminished cardiac performance (by 蠅50%) in all strains of mice. Interestingly, IPC in NCD and HFD Chga‐KO mice induced cardioprotection despite increased ventricular mass and wall thickness in NCD and increased weight gain (>25%) after HFD. Chga‐KO mice differed from CST‐KO mice in being sensitive to insulin even after HFD as assessed by insulin tolerance test and insulin clamp studies, indicating a novel role of insulin sensitivity in cardioprotection. We conclude that the presence of CST and maintenance of insulin sensitivity are two obligatory requirements for achieving cardioprotection from I/R injury through IPC.
The effect of ethanol on loperamide‐induced tolerance (653.15)
Authors: Khan Fayez , Stevens David , Scoggins Krista , Akbarali Hamid , Dewey William
Published: June 2021

We have reported that the mechanism for the production of morphine tolerance differs in the brain and ileum. In more recent studies we have shown that ethanol reversed morphine antinociceptive tolerance in a dose dependent manner. Therefore we investigated the ability of ethanol to reverse the tolerance produced by chronic exposure to Loperamide, a peripherally selective μ‐opioid receptor agonist that inhibits gastrointestinal motility. Gastrointestinal transit was measured by intragastric gavage of a charcoal meal. In vehicle treated mice, the leading edge of charcoal after 30 min transits 74 ± 6 % (n=6) of the length of the ileum. When Loperamide (0.1‐10 mg/kg) was administered subcutaneously 15 minutes before charcoal meal gavage, it significantly and dose‐dependently inhibited gastrointestinal transit. Loperamide (30 mg/kg) given acutely inhibited intestinal transit by 75%. When this dose was administered subcutaneously twice daily for two days and again at 15 minutes before oral injection of the charcoal meal the inhibitory effect on gastrointestinal transit was reduced to 46% (n=6) inhibition; suggesting that tolerance to loperamide occurred. When Loperamide (30 mg/kg) was chronically administered subcutaneous twice daily for two days and on the third day, ethanol (1g/kg) was injected intraperitoneally 30 minutes before a loperamide (30 mg/kg) challenge dose, the inhibition was 47% (n=6) indicating that ethanol did not reverse the tolerance produced by loperamide. These data suggest that ethanol reverses opioid tolerance to central efffects but not to their effects on the ileum.

Grant Funding Source: Supported by NIH DA024009 and ASPET SURF Award (FK)
Role of genistein on EGFR phosphorylation in acute liver damage induce by administration of carbon tetrachloride in rat (653.17)
Authors: Alvarez‐Ayala Elizabeth , Reyes‐Esparza Jorge , Rodriguez‐Fragoso Lourdes
Published: June 2021

Our aim was to study the effect of genistein on EGFR phosphorylation on liver rats with acute damage induces by administration of carbon tetrachloride (CCl4). For this study, we used male Wistar rats which were randomly divided into four groups: control, genistein 5mg/kg oral (2 weeks), acute damage CCl4 4mg/kg i.p. (4 weeks) and acute damage+genistein. Rats were sacrificed and liver were collected for determinate EGFR expression and tyrosine phosphorylation by western blot and immunoprecipitation. EGFR expression was elevated in groups with acute liver damage and acute damage+genistein. On the other hand when we analyzed EGFR phosphorylation we founded a reduction on Tyr845 EGFR phosphorylation in group with liver damage, nevertheless the genistein increased this phosphorylation 5 fold compared with CCl4 group, on Tyr1045 the phosphorylation was to decrease in acute damage and acute damage+genistein group. On Tyr992 and Tyr1068 EGFR phosphorylation genistein and acute damage not showed modification. In conclusion, the genistein can be involved in process like proliferation, apoptosis and EGFR degradation in the liver during acute damage, because it modifies specific tyrosine phosphorylation in EGFR.
Effect of HIV‐1 Tat on enteric neuropathogenesis (653.6)
Authors: Ngwainmbi Joy , De Dipanjana , El‐Hage Nazira , Fitting Sylvia , Dewey William , Hauser Kurt , Akbarali Hamid
Published: June 2021

The gastrointestinal (GI) tract is a major target and reservoir of the HIV virus. Acute and chronic HIV infection induces a higher level of CD4 T cell depletion in the GI tract compared to peripheral blood and affects motility and digestion. Since the enteric nervous system regulates these processes, it may be a potential target of HIV infection. The majority of HIV effects are not achieved by lytic propagation of the virus but by viral proteins. HIV‐1 Tat, a regulatory protein between 86 to 101 amino acids is released by infected cells and together with other HIV proteins has been shown to modulate neuronal function in the central nervous system. However little is known about its effects on the GI tract. In this study, we examined the effects of Tat on single isolated enteric neurons and in isolated ileum tissue. At the cellular level, in whole cell patch clamp experiments, Tat increased enteric neuronal excitability both in Tat treated (n=20) and in ileum neurons isolated from mice expressing the tat gene (n=9) by a significant increase in number of action potentials evoked, decrease in rheobase (from 20pA to 10pA) and threshold potential (from ‐13.2mV to ‐16.1 mV). This increase was mediated by a leftward shift of the sodium channel activation curve (‐ 10mV decrease in V0.5 of activation) thereby increasing the steady state availability of sodium channels. Tat selectively increased the mRNA and protein expression of Nav1.7 and Nav1.8 isoforms of the sodium channel. In isolated ileum tissues, Tat elicited an inflammatory response which was observed by an up regulation of pro‐inflammatory cytokines and increased GI motility thereby contributing to the HIV mediated GI dysmotilities observed in HIV patients

Grant Funding Source: Supported by the National Institute of Health grants DK046367 and DA024009 and the VCU‐IMSD 541332
microRNA 103‐1 exerts a neuroprotective effect in stroke by enhancing ncx1 expression in the brain (654.1)
Authors: Pignataro Giuseppe , Vinciguerra Antonio , Cerullo Pierpaolo , Cuomo Ornella , Formisano Luigi , Guida Natascia , Di Renzo Gianfranco , Annunziato Lucio
Published: June 2021

Background and purpose: MicroRNAs (miRNA) are single‐stranded short RNA molecules that regulate gene expression by either degradation or translational repression of mRNA. Recent studies showed that cerebral ischemia significantly alter cerebral miRNA profiles, mediating profound effect on the disease outcome. In that scenario the Na+/Ca2+ exchanger, by mediating Ca2+ and Na+ fluxes in a bidirectional way across the synaptic plasma membrane, may play a pivotal role in the events leading to anoxic damage. The objective of this study was to set up a valid therapeutical strategy able to contrast the role of specific miRNAs that downregulate NCX expression under experimental conditions mimicking stroke.

Methods: NCX protein expression was evaluated after miRNA and atimiRNA cell trasfection. AntimiRNA 103‐1 was intracerebroventricularly administered in ischemic rat to prevent mir‐103‐1 detrimental action on NCX1.

Results: NCX1 physiological expression was dramatically reduced when cells were treated with mir‐103‐1. Conversely, the in vivo approach showed that antimiRNA‐103‐1 protected from brain ischemia by up‐regulating the neurobeneficial protein NCX1.

Conclusions: The present findings support the idea that blocking mir‐103‐1 by microRNA inhibitors is a reasonable strategy to stop neurodetrimental downregulation of NCX occurring during ischemic conditions.
Doxorubicin inhibits lymphatic function in vitro and in vivo (655.1)
Authors: Stolarz Amanda , Pathan Asif , Versluis Rachel , Fletcher Terry , Stimers Joseph , Galanzha Ekaterina , Rusch Nancy
Published: June 2021

Doxorubicin (DOX) is an independent risk factor for lymphedema in breast cancer patients after surgery and/or radiation therapy. The mechanisms by which DOX contributes to lymphedema are unclear. We hypothesized that DOX directly inhibits lymphatic contractions and lymph flow by a mechanism distinct from its cytotoxic action. Isolated rat mesenteric lymph vessels (LVs) were cannulated, pressurized (4‐5 mm Hg) and equilibrated in a no‐flow system using physiological salt solution (37°C) until spontaneous contractions were stable. An edge‐detection system recorded changes in external diameter. Adding DOX into the bath at increasing concentrations (0.5 to 20 µmol/L; n=7) progressively attenuated the amplitude and area under the curve (AUC) of LV contractions, culminating in reductions of ~55% (amplitude) and ~60% (AUC). Washout of DOX partially restored contractile activity. High‐resolution, intravital optical imaging of rat mesenteric LVs in vivo also revealed that DOX (10 µmol/L) inhibits LV spontaneous contractions and reduces lymph flow (n=2). DOX reduced lymph flow by ~66% (t=10 min) and 100% (t=30 min) from baseline. Thus, DOX may contribute to lymphedema by directly inhibiting LV contractile function as an off‐target effect.

Grant Funding Source: Partially supported by a Rho Chi‐AFPE Graduate Fellowship (AS)
In vitro and in vivo evaluation of doxorubicin‐based anti‐cancer treatment in combination with the herbal medicine black cohosh (655.3)
Authors: Szeffler Agata , Stawarz Andrew , Slania Stephanie , Kalinowski Leszek , Dobrucki Lawrence , Dobrucki Iwona
Published: June 2021

Many breast cancer patients use herbal extracts of black cohosh (BC) in combination with doxorubicin (DOX). In this study we evaluated the synergistic effects of BC and DOX on: (1) in vitro viability, colony formation of human breast cancer cells (MCF‐7), and (2) in vivo evaluation of tumor growth and metabolism.

For viability study, MCF‐7 were incubated with DOX (1‐100μM for 2hrs) and BC (100xdose for 6hrs) alone or combined. To assess the MCF‐7 survival, we used a clonogenic assay. MCF‐7 were incubated with DOX (0.1‐10μM for 2hrs) and BC (10‐100x dose for 6hrs) alone or combined, and cultured for 2wks. Colonies were scanned with a digital stereoscope and counted.

In vivo experiments were performed on OVEX nude mice injected with MCF‐7. After 6wks of tumor growth, animals were divided into groups: a control and those receiving DOX (4mg/kg/wk), BC (20mg/kg/d) or both DOX and BC. Tumor growth was measured twice a week using digital calipers. We performed PET‐CT imaging with 18F‐FDG at baseline, 2 and 6 wks after the treatment. PET‐CT images were analyzed and tumor activity expressed as %ID/g.

We observed significant increase in the number of apoptotic cells after incubation with both DOX and BC vs. control. DOX altered the potential of MCF‐7 cells to form colonies, however co‐incubation with BC did not affect this process. In vivo PET‐CT revealed significant reduction in tumor metabolism after treatment with DOX (40%) and BC (20%) alone. However, we did not observe any synergistic effects when treated with DOX and BC. Both DOX and BC inhibited tumor growth by 20% and 12%, respectively. DOX and BC treatment resulted in a 57% reduction in tumor size.

BC exerts synergistic effects on DOX cytotoxicity resulting in reduction in tumor size, but this change is independent of in vitro ability of MCF‐7 to form colonies, and in vivo tumor glucose metabolism. Further studies are necessary to establish which mechanism underlies these effects and to examine potential clinical implications in management of breast cancer patients.

Grant Funding Source: UIUC Research Board Award (Dobrucki, I.T.)
Vomiting in docetaxel chemotherapy: which to blame, dose or gene? (655.4)
Authors: Jabir Rafid , Ho Gwo , Anuar Muhammad Azrif , Stanslas Johnson
Published: June 2021

The adverse effects (AEs) of docetaxel are major limiting factors of its success in the battle against cancer. AEs can be hematological and non‐hematological (nausea, vomiting, diarrhea, rash and mucositis). Eventually, the dose is often reduced or regimen is diverted. Single nucleotide polymorphisms (SNPs) of genes encoding for proteins involved in the transport of docetaxel (ABCB1 and SLCO1B3) could potentially cause accumulation of the drug and consequent AEs. Therefore, we investigated the association between the SNPs (ABCB1 3435CT and SLCO1B3 334TG) and docetaxel AEs in seventy five Malaysian breast cancer patients receiving docetaxel as a single agent chemotherapy at 90‐100 mg/m2 and 75 mg/m2 in the adjuvant and metastatic settings, respectively. The SNPs were analyzed using PCR‐RFLP technique followed by DNA sequencing of selected samples. The patients were Chinese 37 (49%), Malays 32 (43%) and Indians 6 (8%). We found no significant association between the doses given and development of vomiting. However, Indian patients having ABCB1 3435TT (mutant) had a significant association with the development of vomiting (p=0.03). The SNPs‐AEs association is an essential step in the achievement of personalized docetaxel chemotherapy milestones.

Grant Funding Source: Malaysian Ministry of Higher Education‐FRGS (04‐04‐10‐848FR) and University Malaya HIR‐UM.C/625/1
Glucocorticoid receptor in locus coeruleus mediates the repetitive treatment of corticosterone‐induced sleep disorders (656.6)
Authors: Wang Zi‐Jun , Zhang Xue‐Qiong , Cui Xiang‐Yu , Cui Su‐Ying , Zhang Yong‐He
Published: June 2021

Constantly increased cortisol level resulted from stress can cause serious physical or psychological disorders, but the mechanism of stress‐induced sleep disorders remains unclear. Here we described a novel model, which enables us to investigate stress mimicked sleep disorders induced by repetitive administration of corticosterone (CORT). Rats were treated with CORT for 7 days and the sleep parameters were assessed by EEG. In this model, we observed significant decrease in total sleep time and increase in REM sleep time. Meanwhile, the glucocorticoid receptor(GR) level was notably lowered in Locus coeruleus(LC). We also observed the activation of noradrenergic neurons in LC and the suppression of GABAergic neurons activity in ventrolateral preoptic area(VLPO). Otherwise, no significant changes were found in other sleep regulating nuclei. In addition, we observed remarkable increase of NE in LC, VLPO and hypothalamus and increase of tyrosine hydroxylase in LC and decrease of glutamic acid decarboxylase in VLPO. Microinjection of GR antagonist, RU486 into LC reversed the sleep changes. These results suggested that GR in LC may play a role in the stress mimicked sleep disorders and support the hypothesis that the repetitive treatment of CORT may lower the GR level and induce the activation of noradrenergic neurons in LC, then inhibit GABAergic neurons in VLPO and consequently induced sleep disorders.

Grant Funding Source: Supported by National Natural Science Foundation of China (No. 81173031, 81202511 and 81302746)
Euphol from Euphorbia tirucalli induces human gastric cancer death through the degradation of TGFβR in lipid raft membrane microdomain (657.12)
Authors: Lin Ming‐wei , Huang Yaw‐Bin , Hsieh Chia‐Yuan , Wu Deng‐Chyang , Chang Fang‐Rong , Chen Chun‐Lin
Published: June 2021

Transforming growth factor‐β (TGF‐β) is correlated with poorer prognosis in gastric cancer. Therefore, inhibition of the TGF‐β pathway is an attractive strategy for the treatment of cancer. In this study, we demonstrated that euphol, a euphane‐type triterpene alcohol, attenuated TGF‐β‐induced smad2 phosphorylation, and induced gastric cancer cell death through mitochondrial‐dependent apoptosis. Euphol enriched TGF‐β receptors expression in cell membrane lipid raft microdomain within an hour, and promoted them degradation within 24 hours. The interaction of lipid membrane and euphol was confirmed by liquid chromatography‐mass spectrometry and Biacore‐surface plasmon resonance analysis. In conclusion, our results suggest that euphol may distribute membrane protein TGF‐β receptors in lipid rafts, and these membrane modifications may induce cancer cell death.

Grant Funding Source: Supported by Grant from the Department of Health, Executive Yuan, ROC (DOH100‐TD‐C‐111‐002)
Neuroprotective effects of Lepidium meyenii (Maca) and macamides against amyloid‐beta (25‐35) induced toxicity in B‐35 neuroblastoma cells (657.13)
Authors: Alquraini Ali , Waggas Dania , Bӧhlke Mark , Maher Timothy , Pino‐Figueroa Alejandro
Published: June 2021

Production and deposition of Amyloid‐beta (Aβ) peptide is a hallmark of Alzheimer’s disease. The Maca plant (Lepidium meyenii) has been used as a traditional medicine to treat several diseases. It contains macamides, compounds which are structurally similar to the endocannabinoids which have neuroprotective effects. The purpose of this study was to evaluate the neuroprotective effects of a Maca pentane extract (5‐50 µg/mL) and some of the component macamides: N‐benzylpalmitamide, N‐benzyloleamide and N‐(3‐methoxybenzyl)palmitamide (1‐30 µM) on the B‐35 neuroblastoma cell line. B‐35 cells were pre‐treated with extract or macamides and subsequently exposed to a neurotoxic concentration (10 µM) of Aβ (25‐35). A cell viability assay utilizing 3‐(4,5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium (MTS) was performed to evaluate the neuroprotective effects. The results demonstrated that the Maca extract and N‐(3‐methoxybenzyl)palmitamide counteracted the toxicity produced by Aβ (25‐35), demonstrating significant increases in cell viability (34% and 21% respectively). Furthermore, when the tested compounds were evaluated for antioxidant activity and their effects on caspase 3, neither the Maca extract nor N‐(3‐methoxybenzyl)palmitamide demonstrated an antioxidant effect or caspase 3 inhibition. These results suggest that the Maca extract and macamides may produce neuroprotective effects against Aβ by mechanisms other than caspase 3 inhibition or an antioxidant effect.
Phytochemical study and antinociceptive effect of the hexanic extract of leaves from Combretum duarteanum and friedelin in orofacial nociceptive models (657.14)
Authors: Brito Renan , Quintans Jullyana , Costa Emmanoel , Tavares Josean , Estevam Charles , Barison Andersson , Silva Marcelo , Quintans‐Júnior Lucindo
Published: June 2021

We investigated the antinociceptive effects of the hexanic extract (HE) of leaves from C. duarteanum and friedelin, its mainly compound, in formalin‐, glutamate‐ and capsaicin‐ induced orofacial nociception models. In order to isolate of friedelin from HE, flash chromatography technique was used. Male Swiss mice (n=8/group) were pretreated with HE (100, 200 or 400 mg/kg; p.o.), friedelin (50 mg/kg, p.o.), vehicle (Tween 80 0.2%; p.o.) or morphine (5 mg/kg; i.p.) before the injection of algogen agents into the right upper lip (perinasal area). The test of formalin‐induced orofacial nociception showed that HE and friedelin significantly reduced nociception (p<0.001), in both phases of testing, when compared with the vehicle. In the glutamate and capsaicin‐induced orofacial nociception tests, pre‐treatment with HE produced a significant orofacial anti‐nociception (p<0.001), at all doses tested, when compared with the control group. The results suggest the HE and friedelin possess antinociceptive effects in models of orofacial nociception in rodents.

Grant Funding Source: Suported by CAPES/Brazil, CNPq/Brazil and FAPITEC/Brazil
The marine pharmacology and pharmaceuticals pipeline in 2013 (657.17)
Authors: Mayer Alejandro , Glaser Keith
Published: June 2021

In October 2013 there were six FDA‐approved marine‐derived drugs: for cancer, cytarabine (Cytosar‐U®, Depocyt®, FDA‐approved 1969), eribulin mesylate (Halaven®, FDA‐approved 2010), trabectedin (Yondelis®, FDA‐orphan drug approval 2005) being EU‐registered, and brentuximab vedotin (Adcetris®, FDA‐approved 2011); for pain, ziconotide (Prialt®, FDA‐approved 2004); for hypertriglyceridemia omega‐3‐acid ethyl esters (Lovaza®, FDA‐approved 2004). The clinical marine pharmaceutical pipeline (Mayer et al. TIPS 31:255‐265, 2010) consisted of 10 marine‐derived compounds in clinical development, including three new monoclonal antibodies conjugated to monomethyl auristatin E, a synthetic analog of the marine compound dolastatin, currently in either Phase I, Phase II or Phase III clinical trials. There are currently at least 14 other auristatin‐containing antibody drug conjugates in clinical development. Updated information on the clinical marine pharmaceutical pipeline is available at http://marinepharmacology.midwestern.edu/clinPipeline.htm. Finally, the global preclinical marine pharmacology pipeline was reviewed in Mayer et al. MARINE DRUGS 11:2510‐2573, 2013). In late 2013, both the marine pharmacology preclinical and clinical pharmaceutical pipelines remained very active. Supported by Midwestern University.
Triptolide inhibits MCF‐7 cell growth via estrogen receptor‐α‐related pathways (657.5)
Authors: Sun Lixin , Li Han , Zhang Luyong
Published: June 2021

Triptolide, a diterpenoid triepoxide from the traditional Chinese medicinal herb Tripterygium wilfordii Hook. f., is a potential treatment for autoimmune diseases as well a possible anti‐tumor agent. Our previous studies have shown that triptolide inhibits the proliferation of breast cancer cells in vitro and more potently reduces growth of estrogen receptor α (ERα)‐positive breast cancer cells than ERα‐negative cells. In this study, its ability to block the proliferation of breast cancer and its molecular mechanism of action were investigated. Methods: Cell viability was assessed by PrestoBlue assay. Real‐time PCR and western were used to determine ERα expression. To further investigate the effect of ERα on triptolide inhibition of cell growth, MDA‐MB‐231 cells that overexpress ERα were used. The MCF‐7 xenograft mouse tumor model was used to measure the effects of triptolide on breast cancer in vivo. Results:Levels of ERα and phosphorylated ERK were reduced by triptolide. Overexpression of ERα in MDA‐MB‐231 cells significantly enhanced triptolide’s inhibitory effects. In vivo tumor xenograft model, treatment of triptolide significantly reduced tumor size and ERα expression and inhibited phosphorylated ERK expression. Conclusions:This result suggests that triptolide might be a candidate for prevention of ER‐positive breast cancer cell lines.
Anti‐asthma and anti‐peritonitis effects of O‐methylthioangeloylsencathenone (657.8)
Authors: Im Dong‐Soon , Lee Kyoung‐Pil , Park Soo‐Jin , Kang Saeromi , Noh Dae‐Young , Kim Jung‐Min , Lee Young‐Geun , Choi Young‐Whan
Published: June 2021

To elucidate the anti‐allergic and anti‐inflammatory effects of Petasites genus, we have studied effect of o‐methylthioangeloylsencathenone, a sesquiterpene from Petasites genus on lipopolysaccharide‐induced peritonitis model mouse and ovulalbumin‐induced asthma model mouse. O‐methylthioangeloylsencathenone inhibited antigen‐induced degranulation of β‐hexosamidase in RBL‐2H3 mast cells. In an ovalbumin‐induced mouse asthma model, o‐methylthioangeloylsencathenone inhibited accumulation of eosinophils, macrophages, and lymphocytes significantly in bronchioalveolar fluids (BALF) of asthma mouse. O‐methylthioangeloylsencathenone inhibited also lipopolysaccharide (LPS)‐induced mRNA induction of cyclooxygenase 2 (COX‐2) and inducible nitric oxide synthase (iNOS), detected by RT‐PCR in mouse peritoneal macrophages. O‐methylthioangeloylsencathenone also inhibited LPS‐induced protein induction of COX‐2 and iNOS, detected by Western blotting in mouse peritoneal macrophages. Furthermore, o‐methylthio angeloylsencathenone inhibited production of nitrites, the enzyme product of iNOS in a concentration‐dependent manner in the macrophages. Furthermore, o‐methylthioangeloylsencathenone significantly inhibited migration of neutrophils in the peritoneum in LPS‐induced mouse peritonitis model. In summary, o‐methylthioangeloylsencathenone in Petasites genus acts as an anti‐asthma and anti‐inflammatory constituent.

Grant Funding Source: This research was supported by High Value‐added Food Technology Development (111135‐03‐2‐SB030)
Identification of a novel anti‐inflammatory compound from Schisandra chinesis (657.9)
Authors: Im Dong‐Soon , Lee Kyoung‐Pil , Kang Saeromi , Park Soo‐Jin , Noh Dae‐Young , Kim Jung‐Min , Lee Young‐Geun , Choi Young‐Whan
Published: June 2021

To elucidate the anti‐inflammatory and anti‐allergic effects of Schisandra chinensis, we studied the effect of a novel compound isolated from S. chinensis. The compound M isolated from Schisandra chinensis fruits is a novel sesquiterpine, a derivative of α‐cubebene.

We found that the compound M concentration‐dependently inhibited antigen‐induced degranulation, which was measured in RBL‐2H3 mast cells by measuring β‐hexosamidase activity. The compound M also inhibited the inductions of inducible nitric oxide synthase and cyclooxygenase 2 at mRNA and protein levels, which was measured by Western blotting in mouse peritoneal macrophages. And LPS‐induced productions of nitric oxide and prostaglandin E2 were inhibited by the compound M. The compound M modulated LPS‐induced activations of ERK, JNK, and p38MAPK in mouse peritoneal macrophages. Furthermore, the compound M strongly inhibited the accumulation of neutrophils, eosinophils, and basophils to peritoneal fluid in a LPS‐induced peritonitis model. The compound M has suppressive properties for inflammatory responses and may be utilized as a potent agent for the treatment of inflammatory diseases.

Grant Funding Source: This research was supported by High Value‐added Food Technology Development (111135‐03‐2‐SB030)