bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tridax procumbens suppresses cellular markers of tumour growth, proliferation and invasion in pancreatic ductal adenocarcinoma cell line
Authors: Samuel Ekundayo , Tosh David , Odunola Oyeronke 
Published: April 2020

Pancreatic ductal adenocarcinoma is one of the most fatal solid malignancies, with a 5‐year survival rate of approximately five percent. Hence, there is much interest in developing promising anticancer drugs. To address this issue, we cultured the pancreatic ductal adenocarcinoma cell line (Panc‐1) in supplemented Dulbecco’s Modified Eagle's medium at 37°C in a humidified incubator with 5% CO2. Panc‐1 cells were cultured without or with 20 μg/ml of the dichloromethane fraction (DCMF) of the ethanol leaf extract of Tridax procumbens. Dimethyl sulfoxide was used as control. We examined the expression levels of ductal (cytokeratin 7, peanut agglutinin), mesenchymal (vimentin), cell cycle and proliferation markers (Ki67, p53, p21Cip1/Wap‐1) by immunofluorescence. Confocal images were collected (n≥3) and processed with Image J and Zen 2.3 lite software. Data were analysed using student’s t‐test and results expressed as mean ±standard error of mean with P value <0.05 considered as significant.We observed a significant reduction (p<0.05) in the expression levels of cytokeratin 7, peanut agglutinin, vimentin and Ki67 proteins in DCMF‐treated Panc‐1 cells as compared to control cells. There was a significant increase (p<0.05) in the expression levels of p53 and p21Cip1/Wap‐1 proteins in DCMF‐treated Panc‐1 cells as compared to the control. In conclusion, overexpression of cytokeratin 7, peanut agglutinin, vimentin and Ki67, and reduced expression of p53 and p21Cip1/Wap‐1 in pancreatic cancer correlate with increased tumour growth, invasion and poor prognosis. Therefore, results from this study suggest that Tridax procumbens could be a potential therapeutic agent in the treatment of pancreatic ductal adenocarcinoma.Support or Funding InformationWe appreciate the support and funding received from the University of Ibadan, Ibadan, Nigeria, Commonwealth Scholarship Commission, United Kingdom and University of Bath, United Kingdom.
Cardiovascular Programming by Preeclampsia Sensitizes Mother Rats to Hemodynamic and Cardiac Autonomic Depressant Effects of Postpartum Endotoxemia
Authors: Abuiessa Salwa A. , El-Gowilly Sahar M. , Helmy Mai M. , El-Mas Mahmoud M. 
Published: April 2020

Preeclampsia is a pregnancy related disorder with serious maternal and fetal consequences. Considering that postpartum infection and endotoxemia are important causes of maternal death, we investigated whether preeclamptic maternal programming alters cardiovascular and cardiac autonomic consequences of endotoxemia in rats. Preeclampsia was induced by daily oral administration of 50 mg/kg of L‐NAME for 7 days starting from day 14 of conception. Changes in blood pressure (BP), heart rate (HR), and cardiac sympathovagal balance caused by i.v. administration of lipopolysaccharide (LPS) were assessed in conscious mothers pre‐instrumented with femoral indwelling catheters at 3 weeks (weaning time) and 9 weeks postpartum. At weaning, LPS caused significantly greater falls in BP and rises in HR and LV contractility (dP/dtmax) in PE rats compared with their non‐PE counterparts. Additionally, more apparent reductions in time and frequency domain indices of heart rate variability and shifts in cardiac sympathovagal balance (low‐frequency/high‐frequency ratio, LF/HF) towards parasympathetic dominance were observed after treatment of weaning PE mothers with LPS. Measurement of arterial baroreflex activity by the vasoactive method revealed that LPS attenuated reflex bradycardic responses to phenylephrine in endotoxic PE, but not non‐PE, weaning rats. Remarkably, the intensified cardiovascular effects of endotoxemia in weaning PE rats were found to subside after 9 weeks of delivery. In conclusion, maternal programming by preeclampsia elicits short‐term exacerbation of cardiovascular and autonomic derangements caused by endotoxemia in rats.Support or Funding InformationSupported by the Science and Technology Development Fund, Egypt (STDF Grants No. 14895)
TNFα Decreases Succinate Dehydrogenase Activity in Motor Neurons
Authors: Yap Jane Q. , Heino Amanda , Brown Alyssa D. , Rana Sabhya Denise , Fogarty Matthew J. , Delmotte Philippe D. , Mantilla Carlos B. , Sieck Gary C. 
Published: April 2020

Inflammation triggers a homeostatic response that is mediated by pro‐inflammatory cytokines such as TNFα. We hypothesize that in a motor neuron‐like cell line, NSC‐34, TNFα decreases mitochondrial function, specifically succinate dehydrogenase (SDH) activity, which is a key enzyme in the tricarboxylic acid (TCA) cycle as well as complex II in the electron transport chain (ETC). Experiments were performed on cultured NSC‐34 cells that were differentiated by serum depletion until neurite structures (>50 μm) were observed. The differentiated cells were treated with TNFα (20 ng/ml) for 24 h. To measure the maximum velocity of the SDH reaction (SDHmax), cells were incubated in a solution containing 80 mM succinate and 1.5 mM nitro blue tetrazolium (NBT), the reaction indicator. As the SDH reaction proceeded, images were acquired every 15 s using a 40× objective on an Olympus IX71 inverted microscope. Linearity of the SDH reaction was confirmed across the 8‐min period, and SDHmax was determined and expressed as mM fumarate/L tissue/min. To determine mitochondrial volume density, the cells were loaded with MitoTracker Green and imaged in 3D using a confocal microscope. The cell volume occupied by labeled mitochondria was determined from 3D reconstructions of 1 mm optical slices. In NSC‐34 cells exposed to TNFα, we found that mitochondrial volume density increased, and mitochondria were fragmented as compared to untreated controls. We also found that TNFα exposure decreased SDHmax when normalized for mitochondrial volume density. In other cell types, we found that TNFα‐induces an increase in PGC1a and Drp1 expression and a decrease in Mfn2 expression, all of which affect mitochondrial structure. In related studies, we also found that TNFα exposure decreases O2 consumption per mitochondrion in NSC‐34 cells. We conclude that by reducing the redox potential (SDHmax) of mitochondria, O2 consumption per mitochondrion is reduced, thereby damping ROS formation. The impact of this homeostatic response is offset by mitochondrial biogenesis and an increase in mitochondrial volume density, thereby maintaining overall cellular O2 consumption and ATP production but at reduced cost of ROS formation.Support or Funding InformationSupported by NIH grants AG44615 and HL105355.
Using ENCODE Project Data As Part Of A Course‐Based Undergraduate Research Experience (CURE): Student Perspectives And Feedback
Authors: Conrad William H. 
Published: April 2020

At Lake Forest College, molecular biology is taught as a third‐year upper‐level course. A central learning objective of this course is for students to independently formulate and test hypotheses by evaluating genome‐wide epigenetic data. In this course‐based undergraduate research project, students (1) select high confidence differentially methylated regions (DMRs) present in colorectal cancer samples (2) justify their selection based on literature review (3) predict the effect of DMR methylation on gene expression based on ENCODE project data, (4) predict if the DMR is methylated in HCT‐116 colorectal cancer cells, and (5) test if the DMR is methylated in HCT‐116s and demethylated following decitabine treatment. I will present anonymous survey data of students’ perceptions of the scientific method and their perceptions of themselves as researchers as they engage in research based on big data projects.Support or Funding InformationResearch equipment funded by the Sherman Fairchild Foundation Scientific Equipment Grant. Other funding provided by Lake Forest College Biochemistry and Molecular Biology Program.
Pharmacological Characterisation of PACAP‐responsive Receptors Reveals Signalling Bias and Probe‐dependent Antagonism
Authors: Tasma Zoe , Hay Debbie , Walker Christopher 
Published: April 2020

Pituitary adenylate cyclase‐activating peptide (PACAP) is a neuropeptide which exists in two biologically active forms; PACAP‐27 and PACAP‐38. PACAP is widely expressed in the brain and periphery where it is involved in many physiological processes including pain, inflammation, stress and anxiety. PACAP potently activates three different G protein‐coupled receptors (GPCRs); the PAC1 receptor, the VPAC1 receptor and VPAC2 receptor. The VPAC receptors are co‐receptors for PACAP and vasoactive intestinal peptide (VIP). The PAC1 receptor has several known splice variants in the N‐terminal domain, which include the PAC1n receptor (full N‐terminus) and the PAC1s receptor (21 amino acid N‐terminal deletion). To develop therapeutics targeting the PACAP system, it is essential to understand how these receptors signal and how this signalling is inhibited. Here we pharmacologically characterised the signalling capabilities of PACAP‐responsive receptors and how these responses were blocked by antagonists.The pharmacology of the human PAC1n, PAC1s, VPAC1, VPAC2 receptors were examined in transfected Cos7 cells. Using concentration‐response curves, the ability of PACAP and related peptides to stimulate cAMP, IP1, pAkt, pERK and pCREB responses were measured. Signalling bias was then quantified. The ability of multiple antagonists to block PACAP‐38, PACAP‐27 or VIP‐induced cAMP responses at PACAP‐responsive receptors was then determined.PACAP‐responsive receptors stimulated activation of all signalling molecules measured. However, only limited signalling bias was observed. For example, VIP was 4‐fold biased towards cAMP production over IP1 production at the PAC1n receptor. Interestingly, the PAC1n and PAC1s receptors displayed distinct pharmacology. At the PAC1s receptor, VIP was found to be 20‐fold more potent than at the PAC1n receptor. Probe‐ or agonist‐dependent antagonism was observed at the PAC1n and PAC1s receptors. Both VIP and PACAP‐27 were more potently antagonised than PACAP‐38 by the antagonists PACAP6–38 and M65.The distinct pharmacological profile displayed by the PAC1s receptor, suggests that it can act as a dual receptor for VIP and PACAP under specific conditions. Furthermore, differences in antagonist pharmacology at PACAP‐responsive receptors suggests the effectiveness of blocking a signalling pathway can be influenced by which endogenous agonist is present. These effects have potential implications for the development and effectiveness of drugs targeting the PAC1 receptor.Support or Funding InformationThis research is supported by grants from the Royal Society of New Zealand Marsden Fund (RSNZ) and the Health Research Council of New Zealand (HRC).
Yiqi‐Huoxue Granule suppresses inflammation induced by Lipopolysaccharide in mice by inhibiting the activation of NF‐κB pathway
Authors: WU HONG , Wang Xinzhou , Gao Shuibo , Dai Liping , Wang Zhentao , Han Lihua 
Published: April 2020

ObjectiveTo study the effects of Yiqi Huoxue Granule (YQHX) on inflammatory responses and endothelial injury induced by lipopolysaccharide (LPS) in mice.MethodsMale C57BL/6 mice (8–12 weeks old) were divided into five groups, including control group, LPS group, YQHX group, PDTC (NF‐κB inhibitor) group, and PDTC plus YQHX group. After orally administered with YQHX for 2 weeks or intraperitoneally injected with PDTC for 1 h, LPS was injected into mice for another 6 h, and serums and aortas were collected, respectively. The levels of TNF‐α and IL‐6 in the serum were detected using ELISA. The mRNA expression levels of VCAM‐1, ICAM‐1, PAI‐1 and TF were determined using qRT‐PCR and the protein levels of these factors as well as the phosphorylation levels of NF‐κB pathway were detected using Western blot.ResultsThe levels of TNF‐α, IL‐6, VCAM‐1, ICAM‐1, PAI‐1 and TF in LPS group were higher than those in control group (P <0.01), and the phosphorylation levels of P65 and IκB were also increased significantly (P <0.01). Compared with the LPS group, the levels of TNF‐α, IL‐6, VCAM‐1, ICAM‐1, PAI‐1, TF and NF‐κB pathway phosphorylation were decreased in YQHX group, PDTC group and YQHX plus PDTC group (P <0.05, P <0.01). There was no significant difference among groups of PDTC and YQHX plus PDTC (P >0.05).ConclusionYQHX suppresses the inflammatory responses and endothelial injuries induced by LPS in mice by inhibiting the activation of NF‐κB pathway.Support or Funding InformationThis work was financially supported by grants from the National Natural Science Foundation of China (Grant No. 81473453 & 81673800)
Investigating Oncogenic Roles of Long Intergenic Non‐Protein Coding RNA 461 (LINC00461) in HDAC6‐Mediated Glioblastoma Malignancy
Authors: Wu An-Chih , Yang Wen-Bin , Chang Wen-Chang , Chuang Jian-Ying 
Published: April 2020

Glioblastoma (GBM) is the most complex, difficult‐to‐treat, aggressive, and lethal type of brain tumor. Thus, studies are urgently needed to determine the molecular mechanisms of GBM malignancy and develop novel concepts for treating the disease. According to our previous study, the selective histone deacetylase 1, 2, 6 (HDAC1, 2, 6) inhibitor MPT0B291 prevents the development of temozolomide (TMZ) resistance in GBM cells and prolongs the survival of mice bearing orthotopic GBM xenografts. Although HDACs are known to regulate the balance of lysine acetylation on histones and nonhistone substrates, the role of HDACs in regulating long non‐coding RNAs (lncRNAs) affecting GBM malignancy remains unclear. By performing microarray analysis, we identified an evolutionary conserved lncRNA LINC00461 that was clearly reduced by MPT0B291 in both TMZ‐resistant GBM cells and stem‐like tumorspheres. Additionally, we found that LINC00461 depletion suppressed GBM cell proliferation via G1 phase arrest. Because the miRNA‐411‐5p/topoisomerase II alpha (TOP2A) axis is known as a target of LINC00461 for controlling chromosome segregation during mitosis, we examined the TOP2A levels and verified that both MPT0B291 treatment and LINC00461 silencing caused dramatic decreases in TOP2A expression. Furthermore, we identified the interaction between HDAC6 and subunit 6 of CCR4‐NOT exoribonuclease complex (CNOT6), an acetylation‐activated deadenylase that promotes RNA degradation. In conclusion, this study will increase the understanding of the role of LINC00461 in GBM malignant properties and reveal whether targeting the HDACs/CCR4‐NOT/LINC00461 axis and its downstream effectors with a selective inhibitor is an effective therapeutic strategy against GBMs.Support or Funding InformationThis work was supported by grants from the Ministry of Science and Technology (MOST 106‐6601‐003‐112_15060508367900001 & 107‐6601‐004‐112_15553117427110001).
Altered oligomeric states in pathogenic ALS2 variants associated with juvenile motor neuron diseases cause loss of ALS2‐mediated endosomal function
Authors: Sato Kai , Otomo Asako , Ueda Mahoko Takahashi , Hiratsuka Yui , Suzuki-Utsunomiya Kyoko , Sugiyama Junya , Murakoshi Shuji , Mitsui Shun , Ono Suzuka , Nakagawa So , Shang Hui-Fang , Hadano Shinji 
Published: April 2020

ALS2, the causative gene product for a number of recessive motor neuron diseases, forms a homophilic oligomer and acts as a guanine nucleotide exchange factor (GEF) for the small GTPase Rab5. This oligomerization is crucial for both Rab5 activation and ALS2‐mediated endosome fusion and maturation in cells. Recently, we have shown that pathogenic missense ALS2 variants retaining the Rab5 GEF activity fail to properly localize to endosomes via Rac1‐stimulated macropinocytosis. However, the molecular mechanisms underlying dysregulated distribution of ALS2 variants still unknown. In this study, we clarify the relationship between intracellular localization and oligomeric states of pathogenic ALS2 variants. Upon Rac1 activation, all variants moved from the cytosol to membrane ruffles, but not to macropinosomes and/or endosomes. Further, most wild‐type ALS2 in cultured cells forms tetramers. Importantly, the sizes of an ALS2 complex carrying missense mutations in the N‐terminus of RLD or in‐frame amino acid deletion in the PH domain were shifted toward high molecular weight, whereas the C‐terminal VPS9 domain missense mutant existed as a smaller dimeric or trimeric smaller form. Furthermore, in silico mutagenesis analyses using the RLD protein structure in conjunction with a cycloheximide chase assay in vitro disclosed that these missense mutations led to decrease in the protein stability. These results suggested that disorganized higher structures of ALS2 variants might explain their impaired endosomal localization and the stability, leading to loss of the ALS2 function. Currently, to further clarify the in vivo structure, gel‐filtration analyses of the ALS2 complex in murine tissues, including the brain, spinal cord and liver, are underway.Support or Funding InformationThis work was supported by Japanese Society for Promotion of Science Grants 26290018 and 24650189 and in part by the National Natural Science Foundation of China and JSPS Bilateral Joint Research Project.
Anatomy Faculty Views and Opinions about Curricular Reform at Allopathic Medical Schools in the United States
Authors: Taylor Melissa 
Published: April 2020

IntroductionMedical education in the United States has changed dramatically in the last few decades. Medical schools have moved away from a traditional, didactic, and discipline‐based curriculum, to a more integrative, active, and student‐centered curriculum. While much research has documented the specific changes in medical curriculum, few have researched the perceptions towards these changes.AimThis study examines specific changes that have occurred in anatomical science courses at allopathic medical schools in the United States due to curricular reform and how faculty have reacted to those changes.MethodologyA survey was distributed to anatomical science faculty who teach gross anatomy, microscopic anatomy, and/or neuroanatomy at allopathic medical schools in the United States. The survey included both quantitative and qualitative questions related to the change in content coverage, the change in lecture and lab experience, and faculty perceptions of those changes.ResultsFindings from the survey showed that most anatomical science courses are currently taught in an integrated manner with various active learning pedagogical techniques used in the classrooms. The survey also found that most of the anatomical science courses had their contact hours reduced, leaving much less time for the faculty to teach their students the relevant information. Open‐ended survey responses showed that faculty believed the integration of basic and clinical science information was helpful for their students. However, using Mann‐Whitney U and Kruskal Wallis tests relating faculty perceptions on a Likert Scale to changes in the anatomical science medical curriculum, it was found that faculty whose medical school had undergone recent curricular reform had more negative perceptions about their students receiving adequate instruction in anatomical science material.Discussion and ConclusionThese data provided valuable, though contrasting, information about medical curricular changes and faculty perspectives about those changes. While some faculty had positive things to say about the curricular changes, others believed it was not beneficial to their students. These more negative views should not go unnoticed by administration designing the medical curriculum, and more heed should be taken to listening to all the stakeholders in a medical curriculum.
Chronic Low‐Dose ICV Infusion of Agrp in Mice May Suppress Indices of Resting Metabolism in Brown Adipose Tissue Independent of Effects on Feeding Behavior
Authors: Oliveira Vanessa , Balapattabi Kirthikaa , Patil Chetan N. , Hei Megan Vande , Silva Sebastião D. , Reho John J. , Sigmund Curt D. , Grobe Justin L. 
Published: April 2020

Agouti‐related peptide (Agrp) acts within in the central nervous system to stimulate feeding behavior and suppress energy expenditure, which results in body weight gain. Acute and chronic intracerebroventricular (ICV) administration of recombinant Agrp (0.2 to 1.0 nmol/d) induces hyperphagia in rats (Wissig et al. 2003; Tallam et al. 2004), but studies in mice are lacking. Therefore, we tested the effects of this route and range of infusion doses upon food intake, body mass, and adiposity in wildtype male C57BL/6J mice (Jackson Labs, 10–12 wk). Mice were implanted with chronic indwelling ICV cannulae connected to osmotic minipumps (Alzet) to deliver artificial cerebrospinal fluid vehicle (n=6), or 0.1 nmol/d (n=8) or 1.0 nmol/d (n=6) recombinant Agrp(82–132) peptide for 21 days. Unexpectedly, despite the efficacy of these dosing schedules published in rats, these infusion rates had no effect on food intake, body mass, or overall body composition (by NMR) in the present cohort of mice. Interestingly, interscapular “brown” adipose tissue (BAT) was hypertrophied in mice infused with the lower dose of Agrp (aCSF: 0.07±0.01 mg, Agrp 0.1 nmol/d: 0.10±0.01, Agrp 1.0 nmol/d: 0.09±0.01 (mean±sem); p<0.05 by ANOVA), and this difference remained after correction by body mass or tibia length. Although underpowered to detect statistically significant changes with the limited sample size, physiologically relevant trends toward reduction in the mRNA expression of Ucp1 (0.75 (0.92–0.62) fold of aCSF (±1 sem), n=5), Adrb3 (0.65 (0.74–0.57), n=7), and Dio2 (0.49 (0.8–0.42), n=6) were observed in the BAT from mice receiving 1.0 nmol/d Agrp. We conclude that the effects of ICV Agrp on BAT mass (and possible effects on thermogenic gene expression profiles in BAT) are independent of changes in feeding behavior in male C57BL/6J mice. These findings support species‐specific differences in sensitivity to ICV Agrp, and suggest that Agrp influences resting energy expenditure independently from its effects on feeding behavior.Support or Funding InformationHL134850, HL084207