bluuebunny/crossref_metadata_2025_split huggingface dataset:
Decreased sirt3 expression confers growth advantage in chronic lymphocytic leukemia cells (766.8)
Authors: Yu Wei , Denu John 
Published: June 2021

Chronic lymphocytic leukemia (CLL), the most common leukemia in Western countries, is characterized by the expansion and accumulation of functionally defective B cells. NAD+‐dependent sirtuin 3 (SIRT3) is the major deacetylase within the mitochondrial matrix and has been shown to eliminate reactive oxygen species (ROS) by deacetylating and activating isocitrate dehydrogenase 2 (IDH2) and superoxide dismutase 2 (SOD2). Because SIRT3 levels decrease with age and cancer cells undergo aerobic glycolysis and have higher ROS, we hypothesized that CLL cells express lower levels of SIRT3. Indeed, we found that SIRT3 expression was significantly reduced in 12 primary samples from CLL patients and a number of B cell malignancy lines compared to primary B cells from healthy donors,which is correlated with higher levels of acetylated IDH2 and SOD2, lower IDH2 and SOD2 activities and higher ROS levels in CLL cells. Lastly we demonstrated that SIRT3 overexpression could partially reverse the malignant phenotype in a SIRT3‐deficient Raji cell line. In this first investigation of the role of SIRT3 in hematological malignancies, we have shown that loss of SIRT3 confers a growth advantage to both primary malignant CLL cells and B cell malignancy cell lines consistent with enhanced ROS production and the Warburg effect, whereby cancer cells undergo aerobic glycolysis. Our study suggests that activating the SIRT3 pathway could be considered a novel therapeutic approach for treating B cell malignancies, such as CLL.Grant Funding Source: Supported by University of Wisconsin Carbone Comprehensive Cancer Center (UWCCC)
Decrease in cystathionine‐gamma‐lyase (CSE) activity mediates the impaired H2S homeostasis in U937 monocytes, livers of type 1 diabetic rats and peripheral blood mononuclear cells of type 1 diabetic patients: role of hyperglycemia and hyperketo (767.5)
Authors: Manna Prasenjit , Jain Sushil 
Published: June 2021

H2S is a novel signaling molecule. This study investigated the molecular mechanism that causes impaired H2S levels in diabetes using U937 monocytes, livers from streptozotocin‐treated type 1 (T1D) diabetic rats, and PBMC from T1D patients. T1D is associated with both hyperketonemia (acetoacetate, AA and β‐hydroxybutyrate, BHB) and hyperglycemia. Treatment of monocytes with high glucose (HG, 25 mM) and/or AA (4 mM) increased ROS (44%), and decreased CSE (a major enzyme that catalyzes H2S formation) protein expression (53%) and activity (39%), and reduced H2S levels (45%); however, BHB treatment had no effect (n=4). Signal silencing studies with CSE siRNA (71% knock down) showed a decrease in H2S levels (83%) in monocytes compared to controls. Liver plays a major role in the H2S homeostasis. Livers from T1D rats showed a significantly higher ROS production, lower CSE protein expression (48%) and activity (0.268±0.032 vs 0.175±0.014 nmol/min/mg protein), and lower H2S levels (46%) compared to controls (n=6). Further studies with T1D patients (n=17) also demonstrated a decrease in CSE protein expression (44%) and activity (0.099±0.023 vs 0.044±0.005 nmol/min/mg protein) in PBMC compared to those of age‐matched normals (n=18). This study demonstrates for the first time that both hyperglycemia and hyperketonemia mediate a reduction in CSE activity which can contribute to the impaired H2S signaling in diabetes.Grant Funding Source: Supported by NIH RO1 AT007442 and Malcolm Feist Chair in Diabetes
Cocaine enhances HIV‐1 integration in CD4+ T cells by modulating the epigenetic DNA signatures of host genome (776.3)
Authors: Addai Amma , Pandhare Jui , Mantri Chinmay , Dash Chandravanu 
Published: June 2021

Cocaine, a commonly abused drug among HIV patients, is known to increase HIV‐1 replication in cell cultures, peripheral blood mononuclear cells (PBMCs) and animal models. The goal of this study is to examine the effects of cocaine on HIV‐1 integration. During HIV‐1 replication, the viral genome is reverse transcribed into a dsDNA by HIV‐1 reverse transcriptase. This viral dsDNA is transported into the nucleus in the form of a pre‐integration complex (PICs) and integrates into the host genome. Our data demonstrates that cocaine treatment enhances HIV‐1 integration in primary CD4+ T cells. To understand the mechanism, we examined global DNA methylation in cocaine treated cells. Our data revealed that cocaine induced global DNA de‐methylation in CD4+ T cells. Since integration has been suggested to be disfavored in methylated DNA targets, we hypothesize that cocaine enhances integration by reducing the global DNA methylation signature of the host genome. Our in vitro studies using PICs from infected cells illustrate that integration is reduced in target DNA that are methylated compared to unmethylated target. To the best of our knowledge, this is the first report describing a role of cocaine on HIV‐1 integration in activated CD4+ T cells. We believe understanding the mechanism by which cocaine modulates HIV‐1 integration may aid in the discovery of novel targets to cure the co‐morbid condition of drug abuse and HIV/AIDS.Grant Funding Source: *Amma Addai is supported by T32 grant #2T32H007735‐17 from NIH/NHLBI to Dr. S. E. Adunyah
Evaluation of mRNA extraction methods from murine skin (776.4)
Authors: Neumann Drexel , Atwood James , Nasti Tahseen , Tsuruta Yuko , Cochran Barry , Timares Laura 
Published: June 2021

Intro: Transcriptome research has become increasingly popular to monitor global changes in gene expression. In particular, transcriptome differences between diseases versus healthy states can provide valuable insight into understanding immune response mechanisms. However, the isolation of high‐quality mRNA from tough tissues, such as skin, can represent a significant challenge in research. Herein, we compare two methods for mRNA extraction from two murine skin types with a specific focus on mRNA yields and integrity.Methods: 1cm2 full‐thickness mouse skin (epidermal and dermal layers) samples were freshly harvested, separated from fat, and kept on ice until homogenization. Samples were cut into fragments and placed in 1ml Trizol, then homogenized using a manual Dounce homogenizer on ice, or an automated Omni Bead Ruptor 24 (BR24) bead mill homogenizer without refrigeration. Clarified sample supernatants were mixed with EtOH to precipitate nucleic acids, and sample yields and quality were determined by spectrophotometry. The mRNA purity and integrity of each sample was additionally determined using an Agilent BioAnalyzer.Results: Spectrophotometric data indicated that, depending on skin tissue type and sample size, mRNA yield from samples processed by BR24 versus Dounce was increased 2‐6 fold. The 260/280 ratios were equivalent across both methods; however, the Alginet electropherograms revealed that Dounce homogenization produced samples with unacceptably high MW contaminant DNA and low MW degradation species, while the BR24 processing produced superior mRNA samples of outstanding purity and RNA integrity (RIN). (Omni BR24 RIN values 8.4 ‐ 8.8 ; Dounce RIN, 蠄 4.3). Thus, high quality mRNA can be efficiently isolated from difficult to disrupt tissues such as skin.
Gene expression alterations in patients with post‐traumatic stress disorder (776.6)
Authors: Muhie seid , Yang Ruoting , Hammamieh Rasha , Jett Marti 
Published: June 2021

We profiled transcriptome changes in patients with post‐traumatic stress disorder (PTSD). Whole blood samples from 98 subjects (49 PTSD‐positives and 49 age, race and gender‐ matched controls) were assessed using microarray and real time polymerase chain reaction assays. About 1500 genes were differentially regulated in PTSD‐positive patients compared to the controls. Down‐regulated transcripts were involved in T‐cell immunity, cardiovascular activity, and synaptic transmissions. Suppressed pathways and functions were critical in maintaining one’s sense of well‐being, protective immunity, and cardiovascular health. Induced functions and pathways (significantly associated with up‐regulated transcripts) were largely related to traumatic stress responses such as endoplasmic reticulum stress, DNA repair, wound healing, and synaptic plasticity. Our data shows the pervasive negative impact of PTSD at the molecular level in regulation of key pathways and functions important to health.Grant Funding Source: US Army and Veteran's Affairs
Comparison at the peptide level with post‐translational modification consideration reveals more differences between two unenriched samples (778.1)
Authors: Yin Jianrui , Shao Chen , Jia Lulu , Gao Youhe 
Published: June 2021

In shotgun strategy, peptide sequences are first identified from MS/MS spectra, and the existence and abundance of the proteins are then inferred from the peptide information. However, the protein inference step can produce errors and a loss of information. To identify the information that is lost using the traditional approaches, this study compared the proteomic data of two leukemia cell lines (Jurkat and K562) at peptide level with consideration of post‐translational modifications (PTMs). After database search, the observed modification name was matched with the modification classification on the Unimod website. Only the peptides with ‘post‐translational’ modifications were compared between the two cell lines. As a result, a total of 44887 non‐redundant peptides were identified in both the Jurkat and K562 cell lines. Of these peptides, even without specific PTM enrichment, 11.43% of them (with at least two spectra in one cell line) existed in different PTM forms between the two cell lines, and 1.89% of the peptides were modified in both cell lines, but with different modifications or on different sites. The sensitivity of PTM identification will become greater and more differences will be found if PTMs are enriched or if the peptides can be compared in a quantitative manner. Therefore, comparing proteomic data at the peptide level with consideration of PTMs can reveal differences between two unenriched samples.Grant Funding Source: Supported by National Basic Research Program of China (2012CB517606, 2013CB530805, 2011CB964901).
Pyroprinting: novel pyrosequencing‐based method for studying E. coli diversity and microbial source tracking (779.8)
Authors: Kent Jason , Alvarado Maria , VanderKelen Jennifer , Montana Aldrin , Soliman Jan , Dekhtyar Alex , Goodman Anya , Kitts Christopher , Black Michael 
Published: June 2021

We have developed a novel genotypic fingerprinting method for typing Escherichia coli strains via simultaneous, multi‐locus pyrosequencing of the internal transcribed spacer (ITS) regions in ribosomal RNA operons. The data generated, called pyroprints, are unique to the polymorphic ITS alleles and their relative frequency among the seven rRNA operons in each E. coli isolate. Pyroprinting is a rapid, inexpensive, and highly reproducible method that produces readily digitized data for computational analysis and effectively discriminates closely related strains. A pyroprint database (www.cplop.org) and analytical tools for environmental forensics and microbial source tracking (MST) applications have been developed. Our pyroprint library currently contains over 5,000 E. coli isolates from avian and mammalian hosts. Pyroprinted isolates have been clustered into strains, some of which are shared by multiple hosts, while others appear host‐specific. To validate this method for MST, we have simulated environmental water samples by spiking them with fecal material from known hosts in pre‐determined ratios. Pyroprinting E. coli isolates from the simulated samples has allowed us to identify the origin of the isolates and ratios of different hosts that contributed to the mixed samples. This new MST method has been employed to identify sources of fecal contamination in San Luis Obispo Creek.Grant Funding Source: Supported by NSF TUES # 1140828.
The α‐arrestin ARRDC3 regulates ubiquitin‐independent ALIX‐mediated GPCR lysosomal sorting (783.2)
Authors: Dores Michael , Lin Huilan , Buck Hannes , Mendez Francisco , Trejo JoAnn 
Published: June 2021

Endosomal sorting and lysosomal degradation of G‐protein coupled receptors (GPCRs) maintains cell homeostasis by attenuating receptor signaling and desensitizing cells to persistent activation. GPCR lysosomal sorting defects cause aberrant cellular responses that contribute to many diseases, including chronic inflammation and cancer metastasis. Here, we demonstrate that the human α‐arrestin ARRDC3 regulates the lysosomal degradation of protease‐activated receptor‐1 (PAR1), a GPCR for thrombin. PAR1 belongs to a subset of GPCRs, including the purinergic receptor P2Y1, that are sorted to lysosomes independent of receptor ubiquitination. Instead, PAR1 and P2Y1 are targeted to lysosomes by YPXnL motifs that are bound by the ESCRT adaptor protein ALIX. ARRDC3 is required for PAR1 degradation, but not for the degradation of ubiquitinated receptors. ARRDC3 regulates ALIX interaction with PAR1 and the ESCRT‐III protein CHMP4. Intriguingly, depletion of ARRDC3 or ALIX blocks recovery of thrombin‐induced dephosphorylation of Yap1, an effector of the Hippo cell proliferation pathway, suggesting that GPCR degradation is important for regulation of Hippo signaling. This study reveals a novel role for α‐arrestins in the ubiquitin‐independent, ALIX‐mediated endosomal trafficking of GPCRs, and demonstrates the importance of lysosomal sorting in the regulation of GPCR‐mediated proliferative signaling.Grant Funding Source: Supported by NIH NIGMS R01 GM090689
The anti‐diabetic drug metformin perturbs proliferation and cellular differentiation of pancreatic cancer cells: a key mechanism of its anticancer activity (784.6)
Authors: Das Amlan , De Archana , Maity Gargi , Banerjee Snigdha , Banerjee Sushanta 
Published: June 2021

Background : Pancreatic cancer is the 5th most common cancer in the United States and is currently the fourth leading cause of cancer death, with a 5‐year overall survival of only 5%. It has been reported that Type 2 diabetes mellitus and associated hyperglycemia, hyperinsulinemia, and inflammation, are known to play important roles in the development of diabetes‐associated pancreatic cancer. Use of the anti‐diabetic drug metformin has been reported to reduce the risk of pancreatic cancer in diabetics and recognized as an antitumor agent with the potential to inhibit the progression of pancreatic tumors. However, the mode of action of Metformin on regulation of cellular differentiation, which are associated with are invasion and metastasis of pancreatic cancer, is uncertain.Objective: The objective of the present study is to determine if Metformin plays any role cellular de‐differentiation through the reprogramming of EMT process that eventually prevents invasive phenotypes associated with CXCR‐4/SDF‐1 pathways.Results: In our study we have observed that treatment of the pancreatic cancer cells such as Panc‐1 and AsPc‐1 resulted in a dose‐dependent inhibition of proliferation, accompanied by the induction of cytotoxicity through the regulation of the tumor suppressor gene p27. Moreover, Metformin treatment resulted in the morphological alteration as well as induction of EMT in pancreatic cancer cells. Metformin treatment prevents in vitro invasion of PC cells to words SDF‐1, a ligand for CXCR4 chemokine receptor actively associated with cancer metastasis.Conclusion: The widely used anti‐diabetic drug metformin can suppress pancreatic tumor progression by inhibiting the chemokine‐induced invasiveness and motility.
Glycomics analysis of resting and activated human platelets (789.1)
Authors: Azadi Parastoo , Ishihara Mayumi , Kumagai Tadahiro , Stalnaker Stephanie , Zhang Hanrui , Park Seongha , Wells Lance , Tiemeyer Michael , Reilly Muredach 
Published: June 2021

Cardiovascular diseases are the most common causes of death in developed countries and their prevalence rate is rapidly growing in developing countries. Genome‐wide association studies (GWAS) have identified associations between ABO blood group status and HLBS (heart, lung, blood, and sleep) disorders. ABO glycans are expressed on glycoproteins and glycosphingolipids (GSLs) of cells and tissues relevant to HLBS disorders,including endothelial cells and platelets. Several major HLBS disorders, particularly acute myocardial infarction and acute lung injury, are characterized by endothelium dysfunction and platelet‐driven thrombosis. Therefore, a comprehensive understanding of the disposition of ABO glycan expression by endothelial cells and platelets is essential for elucidating the mechanisms by which these glycans influence risk for cardiovascular disease. Toward this end, we have undertaken the structural characterization of GSL, and N‐ and O‐linked glycoprotein glycans in resting and activated platelets. The preparations of GSLs are permethylated and analyzed by direct infusion into an NSI‐LTQ/Orbitrap mass spectrometer. N‐ glycans are enzymatically released and O‐glycans are released by reductive beta‐elimination from glycoprotein preparations prior to permethylation and subsequent quantitative analysis by mass spectrometry. For GSLs, expression levels of lactosylceramide (LacCer) and of the sialylated form of LacCer, ganglioside GM3, were significantly decreased and increased, respectively, upon activation. For N‐glycans, high‐mannose structures and mono‐ and disialylated N‐glycans were significantly increased in activated platelets compared to resting platelets. These results are consistent with a systemic upregulation in human platelet glycoconjugate sialylation during activation of platelets and enhanced glycan‐mediated platelet adhesion and thrombosis.Grant Funding Source: NIH
Glycogenome signatures in complex cardiometabolic disease (789.4)
Authors: Ballantyne Rachel , Qu Liming , Liu Jim , Nairn Alison , Li Wenyuan , Azadi Parastoo , Zhang Hanrui , Wells Lance , Tiemeyer Michael , Moremen Kelley , Christie Jason , Li Mingyao , Zhou Xianghong , Reilly Muredach 
Published: June 2021

Cardiometabolic diseases (CMDs) are a leading cause of death. Because of the fundamental roles of glycogenes in cell interactions and tissue remodeling, we hypothesized that a focused interrogation of the human glycogenome in genome wide association studies (GWAS) for CMDs would reveal a role for previously unrecognized glycogenes. We defined a set of 723 human glycogenes, and annotated ~45,000 single nucleotide polymorphisms (hg 19, dbSNP build 137) into these genes, from 16 GWAS datasets for CMD traits, including coronary artery disease, plasma lipids and type 2 diabetes. At a false discovery rate of 5%, we identified 788 SNPs in 83 genes associated with one or more of 11 traits. Of these, 334 SNPs in 36 glycogenes were also significant at a Bonferroni‐adjusted p < 0.05 (unadjusted p < ~10‐6). A weighted score of SNP associations across traits revealed several glycogenes with pleiotropic associations, including COL4A3BP, ABO, PIGU, ST3GAL4, GCK, SDCBP, G6PC2, and FUT2. Our analyses suggest a role for multiple glycogenes in diverse CMDs with specific glycogenes contributing to multiple traits. Functional studies and therapeutic targeting of specific glycogenes and pathways in CMD are warranted.Grant Funding Source: Supported by the National Institutes of Health (U01‐HL108636 and K24‐HL10763)
Production of neuraminidase by Pseudomonas aeruginosa (790.3)
Authors: McClinton Anna , Cioffi Donna , Cioffi Eugene 
Published: June 2021

Rationale: Neuraminidase (NA), an enzyme which cleaves sialic acids from underlying glycans, is considered a virulence factor for several pathogenic bacteria. The opportunistic pathogen Pseudomonas aeruginosa is also known to produce NA. P. aeruginosa NA (PaNA) has been implicated in biofilm formation and colonization of respiratory epithelium in cystic fibrosis. However, a clear role for PaNA in other disease states has not been determined. In order to investigate potential roles of PaNA in infection, the NA enzyme must be isolated and purified. NA production by other bacteria, such as Streptococcus spp, correlates with the late logarithmic or early stationary phases of bacterial growth. Additionally, conditions such as growth media have been shown to influence NA production. In order to optimize production of NA by P. aeruginosa strain Pa103, growth curves were performed along with NA activity assays. Methods: P. aeruginosa strain Pa103 was grown in various broth cultures: Brain Heart Infusion (BHI), BHI +10% Newborn Calf Serum (NCS), and BHI + 10% heat‐inactivated NCS. Samples were taken hourly for NA activity assays. Results: NA activity peaked in the late logarithmic/early stationary phase of growth, regardless of serum added to the media. NA activity was not enhanced by the addition of serum. Conclusion: P. aeruginosa Pa103 produces NA in agreement with patterns observed in other NA producing bacteria.Grant Funding Source: Supported by R01HL107778
Targeting beta‐arrestins in the treatment of Parkinson's disease (796.6)
Authors: Urs Nikhil , Bezard Erwan , Caron Marc 
Published: June 2021

Parkinson’s disease (PD) is a neurodegenerative disorder caused by the progressive loss of dopamine input to the striatum. Dopamine (DA) is a catecholamine neurotransmitter that is released by midbrain DA neurons and activates G‐protein coupled receptors (GPCRs) on postsynaptic striatal neurons. DA GPCRs are known to signal via both G‐protein‐ and beta‐arrestin2 (βarr2) dependent mechanisms and both pathways either individually or in combination, play a major role in control of movement. PD is characterized by severe locomotor deficits and is commonly treated with L‐DOPA, but its prolonged usage causes dyskinesias referred to as L‐DOPA induced dyskinesias (LIDs). Recent studies in animal models of PD have suggested that dyskinesias might be associated with G‐protein mediated signaling through DA receptors. Therefore, preventing G‐protein signaling in PD therapy may be an ideal way to eliminate associated dyskinesias. β‐arrestins have the unique ability to desensitize G protein signaling and at the same time independently activate signaling, thereby mediating locomotion. Our objective is to test the effect of over‐expression or deletion of βarr2 on L‐DOPA induced locomotion and dyskinesia potential in a rodent and non‐human primate PD model. Our results in the rodent model suggest that while the absence of βarr2 reduces the beneficial locomotor response of L‐DOPA, it enhances the manifestation of dyskinesias, presumably because it shifts the system to excessive G protein‐dependent signaling. As expected, in both the rodent and NHP model, over‐expression of βarr2 enhances the efficacy of forward locomotion (i.e on‐time) and at the same time reduces the manifestation of LIDs. Thus increasing activation of βarr2 either pharmacologically or genetically during L‐DOPA therapy may be an ideal way to overcome the motor deficits of Parkinsonian patients and ameliorate undesirable side‐effects of drug therapy.Grant Funding Source: Supported by the Michael J. Fox Foundation for PD research