bluuebunny/crossref_metadata_2025_split huggingface dataset:
PMPMEase inhibitors as potential therapies for castration‐resistant prostate cancer (585.7)
Authors: Poku Rosemary , Amissah Felix , Duverna Randolph , Byron Aguilar , Lamango Nazarius 
Published: June 2021

Castration resistance continues to pose major challenges in the management and treatment of prostate cancer. Monomeric G‐protein signaling is critical in prostate carcinogenesis. These proteins are polyisoprenylated and regulated by polyisoprenylated methylated protein methyl esterase (PMPMEase).It was hypothesized that PMPMEase regulates cell viability and migration. The purpose was to determine the effect of PMPMEase inhibition on the malignant characteristics of prostate cancer cells.The effects of PMPMEase inhibition on cell viability, proliferation, migration and gene profiling were determined with the specific PMPMEase inhibitors, L‐28 and NSL‐RD‐036.PMPMEase specific activities were 1.5 and 4.5‐fold higher in androgen‐dependent 22Rv1 and LNCaP cells and 1.5‐ and 9.8‐fold higher in androgen‐independent DU 145 and PC‐3 prostate cancer cells compared to normal WPE1‐NA22 prostate cells. Treatment of the cells for 48 h with L‐28 induced apoptosis with EC50 values ranging from 1.8 to 4.6 µM. The PMPMEase activity in the treated cells followed a similar profile with EC50 values from 2.3 to 130 µM. L‐28 treatment significantly disrupted F‐actin cytoskeleton in PC‐3 cells and inhibited cell migration. NSL‐RD‐036 inhibited PC‐3 proliferation with an EC50 of 2 μM. The expression of cell proliferation, metastasis, apoptosis and DNA synthesis and repair genes were significantly altered by specific PMPMEase inhibition.PMPMEase inhibition induces apoptosis, disrupts F‐actin and suppresses the migration of androgen‐independent prostate cancer cells. Inhibition and measurement of PMPMEase has the potential to serve in the effective management of aggressive prostate cancer.Grant Funding Source: Supported by National Center for Research Resources and the National Institute of Minority Health an
Altering substrate specificity of aldehyde dehydrogenase 3A1 to enhance acetaldehyde metabolism, in vivo (585.9)
Authors: Chen Che‐Hong , Cruz Leslie , Mochly‐Rosen Daria 
Published: June 2021

Acetaldehyde is a well‐know cytotoxin, and carcinogen. Detoxification of acetaldehyde, which is mainly carried out by the mitochondrial aldehyde dehydrogenase 2 (ALDH2), is crucial to human health. High levels of acetaldehydes are present in the saliva and in circulation after ethanol ingestion and likely contribute to the high incidence of esophageal cancer in alcoholics and in carriers of a common ALDH2 mutation (ALDH2*2) found in 540 million East Asians. Acetaldehyde is also partly responsible for behavior impairment following ethanol consumption. Therefore, means to accelerate acetaldehyde clearance will be beneficial for human health. We previously identified an activator of ALDH2 that accelerated acetaldehyde metabolism. Here we describe the recruitment of an additional ALDH isozyme, ALDH3A1, to accelerate the clearance of acetaldehyde by the use of a small molecule. ALDH2 is the main isozyme that metabolizes acetaldehyde in liver. In contrast, ALDH3A1, which is present in the stomach and mucosa of the esophagus, normally metabolizes aromatic aldehydes and is ineffective in metabolizing acetaldehyde. We identified a small molecule activator, Alda‐89, which increased the Kcat of ALDH3A1 for acetaldehyde and enhanced its catalysis. We showed that when given together with an ALDH2‐specific activator, Alda‐1, Alda‐89 induced a rapid reduction in blood ethanol and acetaldehyde levels after acute ingestion of ethanol in mice. Importantly, treatment with Alda‐1 and Alda‐89 also reduced behavioral impairment in both wild‐type and ALDH2*2 mice. The ability to recruit an enzyme to metabolize a substrate that it usually does not metabolize using a pharmacological agent may represent a novel strategy to increase metabolism of toxic agents, in vivo.Grant Funding Source: Supported by NIH AA11147 to DM‐R
Isolation and characterization of two cDNAs expressed in the Harderian gland of the anole lizard (Anolis carolinensis) (589.4)
Authors: Prokopiak Zoey , Steglich Carolyn 
Published: June 2021

The Harderian gland (HG) is a secretory gland in the eye of most land animals. Its structure has been studied, but little is known about the gland's actual function, which varies depending on species. Structural observations have suggested multiple possible functions (lubrication of the eye, sensing of chemical signals via the vomeronasal organ, or an immunological role) but have little supportive experimental data. To investigate the HG’s function in an organism such as the Anole lizard (Anolis carolinensis), cDNAs were isolated from the HG and sequenced. Two cDNAs (E1 and M1) that code for proteins of unknown structure or function (no significant hits in a GenBank search) were selected for further study. Using RT‐qPCR (reverse transcriptase quantitative polymerase chain reaction), the expression of the cloned genes (cDNAs) that had been isolated from the HG of the Anole lizard were compared to other known Anolis genes (in this case, genes encoding ribosomal proteins) that are expressed in all cells. The E1 cDNA is expressed at high levels in the HG but also expressed at background levels in liver and tongue tissues. The M1 cDNA is expressed at extremely high levels only in the HG. These cDNAs have been subcloned into the pET‐22 expression vector for expression in bacterial cells to provide an unlimited source of the proteins. The functional characterization of the proteins can then be directly investigated.The genome of the Anole lizard was completed in 2010 but is currently incompletely annotated. BLAST searches of the Anole's genome with the E1 and M1 sequences have returned some short matches for the sequences, which locates each of them to a particular scaffold in the genome, but no predicted genes are annotated for those regions. Additional alignment work with MegAlign locates the likely exons and introns of these genes more precisely, thereby providing additional annotation to those regions of the Anole genome.Grant Funding Source: Supported by a Faculty/Student Research Grant and a Student Research Grant from Slippery Rock Univ.
Mutations of the LYL1 gene in patients with T‐cell acute lymphoblastic leukemia (59.7)
Authors: Chan Tsz Yin , Zohren Fabian , Ballard Brandon , Goodell Margaret , Rabin Karen , Lukov Georgi 
Published: June 2021

The lymphoblastic leukemia ‐ 1 (LYL1) gene is a transcription factor believed, when upregulated, to support the formation of T‐cell Acute Lymphoblastic Leukemia (TALL). One possible cause for the upregulation of LYL1 is the presence of mutations, affecting the normal expression, processing or degradation of LYL1. The objective of our study is to identify if there are mutations in the LYL1 gene in patients with TALL, and determine the effect of such mutations on LYL1 expression. RNA, for quantitative PCR, and DNA, for LYL1 sequencing, were extracted from blood and bone marrow samples from twenty TALL patients. Ten samples showed high and intermediate LYL1 expression. Five of them had single nucleotide polymorphism with three of them being non‐silent. LYL1 variants with single or combined mutations were constructed, and their expression, in cultured CHO‐K1 and H441, was measured by western blotting. The LYL1 variants showed significantly increased expression, up to 3.6 fold, compared to the wild type form. Our findings suggest that these mutations may have a role in the expression and processing, or the degradation of LYL1, which not only adds to our understanding of the LYL1 biology, but it may offer one more clue why LYL1 is upregulated in some forms of TALL.Grant Funding Source: BYUH Mentored Research & Student Research Associateship Funds
Elevated plasma biomarkers of chronic inflammation in Gulf War illness (591.3)
Authors: Johnson Gerhard , Slater Billie , Leis Linda , Bach Ronald 
Published: June 2021

As many as 200,000 veterans of the 1990‐1991 Gulf War suffer from unexplained chronic multisymptom illnesses collectively known as Gulf War Illness (GWI). Symptoms of GWI may include muscle and joint pain, unexplained fatigue, and cognitive impairment. The etiology of GWI is undetermined, and no effective treatment has been defined.Objective: To determine if blood abnormalities exist in veterans with GWI that could lead to the identification of potential therapeutic targets.Methods: Eighty‐five gulf war veterans were enrolled. Subjects with GWI were identified by standard (CDC‐10) criteria. Data derived from subjects with GWI (58) were compared to those without GWI (27). Multi‐Analyte Profiling of plasma proteins was performed and analyzed by the Mann‐Whitney rank sum test.Results: Five of 89 proteins were significantly elevated (p<0.05) in the GWI subjects: C‐reactive protein (CRP) (1.75‐fold), leptin (1.67‐fold), interleukin‐1 beta (1.66‐fold), brain‐derived neurotropic factor (1.59‐fold), and matrix metalloproteinase‐9 (1.14‐fold). The 14 individuals in this study with the highest CRP levels, as much as 10‐fold above the median, were all in the GWI group.Conclusion: The biomarker evidence is consistent with activation of the innate immune system. It supports the hypothesis that chronic inflammation is a component of GWI pathophysiology.Grant Funding Source: Supported by VA ORD and DoD CDMRP
Portal bile acid perfusion bypasses ileal FGF19 and suppresses hepatic CYP7A1 expression in rabbits (605.6)
Authors: Xu Guorong , Shang Quan , Guo Grace , Shi Daniel , Saumoy Monica , Salen Gerald 
Published: June 2021

This study challenged the hypothesis that CYP7A1 expression is regulated by a gut‐hepatic signaling pathway FGF15/19‐FGFR4 initiated from the ileum but not the liver. We would like to show that increase of portal bile acid flux alone should be able to suppress CYP7A1 expression. Methods: We developed a rabbit model to infuse glycodeoxycholic acid (GDCA) via the splenic vein to bypass ileal FGF19. Four study groups of rabbits (n=8/group): Control; bile fistula (BF); BF+GDCA (by portal perfusion); and BF+ GDCA‐f (by femoral perfusion). Results: Portal GDCA perfusion in the BF+GDCA group for 6 hours resulted in significant reduction in CYP7A1 mRNA but femoral perfusion in the BF+GDCA‐f group showed no change with the same perfusion rate of GDCA. In both GDCA perfusion groups, there was no increase in ileal FGF19 expression nor portal FGF19 protein levels, but similar increase in biliary bile acid outputs. Conclusions: This study demonstrated that portal bile acid flux alone inhibited CYP7A1 expression without increase of FGF19 in the ileum. However, the same bile acid flux through the liver via the hepatic artery (femoral perfusion) did not suppress the CYP7A1 expression. We propose that bile acid flux through the portal venous system may be the so called “intestinal factor” that suppresses CYP7A1 expression.Grant Funding Source: VA Merit Review Grant
Inhibition of glycosphingolipid synthesis ameliorates atherosclerosis and arterial stiffness in Apo E‐/‐ mice and rabbits fed a high fat and cholesterol diet (607.11)
Authors: Amuzie Christine , Mishra Sumita , Bedja Djahida , Avolio Alberto , Kass David , Berkowitz Dan , Renehan Mark , Chatterjee Subroto 
Published: June 2021

Objective ‐‐ Glycosphingolipids are integral components of the cell membrane and have been shown to serve as messengers, transducing growth factor initiated phenotypes. Here we have examined whether inhibition of glycosphingolipid synthesis could ameliorate atherosclerosis and arterial stiffness in transgenic mice and in rabbits.Methods and Results ‐‐ Apolipoprotein E‐/‐ (apoE‐/‐) mice (12 weeks of age, n = 6) were fed regular chow or a western diet (1.25% cholesterol, 2% fat). Mice were fed 5mg/kg (mpk) or 10 mpk of an inhibitor of glycosphingolipid synthesis inhibitor D‐threo‐1‐phenyl‐2‐decanoylamino‐3‐morpholino‐1‐propanol (D‐PDMP) solubilized in vehicle (5% Tween‐80 in PBS) and the placebo group received vehicle only. At 20 weeks and 36 weeks of age, serial echocardiography was performed without sedation to measure aortic pulse wave velocity (PWV). Feeding mice a western diet markedly increased aortic PWV, aortic intima medial thickening (IMT), oxidized LDL, Ca2+ deposits, and glucosyl‐ and lactosylceramide synthase activity. These were dose‐dependently decreased by feeding D‐PDMP. In liver, D‐PDMP decreased cholesterol and triglyceride levels by raising the expression of SREBP‐1, LDL‐r, HMGCo‐A reductase, and cholesterol efflux genes (e.g., ABCG5, ABCG8). D‐PDMP affected VLDL catabolism by increasing the gene expression for LPL and VLDLr. Rabbits fed a western diet for 90 days had extensive atherosclerosis accompanied by a 17.5‐fold increase in total cholesterol levels and a 3‐fold increase in lactosylceramide levels. This was completely reversed by feeding D‐PDMP.Conclusions ‐‐ Inhibition of glycosphingolipid synthesis ameliorates atherosclerosis and arterial stiffness in ApoE‐/‐ mice and rabbits. Thus, inhibition of glycosphingolipid synthesis may be a novel approach to treating atherosclerosis and arterial stiffness.Grant Funding Source: Supported by P01HL107153
Inhibiting glycosphingolipid glycosyltransferase activity prevents cardiac hypertrophy in apoE‐/‐ mice fed a western diet (607.12)
Authors: Bedja Djahida , Mishra Sumita , Amuzie Christine , Kass David , Avolio Alberto , Chatterjee Subroto 
Published: June 2021

Background: Although diet enriched in high fat and cholesterol can induce cardiac hypertrophy, little is known about the role of glycosphigolipids and glycosyltransferases contributing to this phenotype. Herein, using transgenic apo E‐/‐ mice fed a high fat and cholesterol diet we show that treatment with a glycosphingolipid inhibitor blunts cardiac hypertrophy.Methods and Results: Male apoE‐/‐ mice were fed a western diet (20% fat and 1.25% cholesterol) from 12 weeks of age till 36 weeks with and without D‐threo‐1‐phenyl‐2‐decanoylamino‐3‐morpholino‐1‐propanol (D‐PDMP) daily by oral gavage (vehicle alone, 5 mpk and 10mpk) . At 12 week, 20 week and 36 week, trans ‐thoracic echocardiography were performed in conscious mice using an ultrasound system (n=5 in each group). Mice were euthanized once maladaptive cardiac hypertrophy time point is reach. Tissues such as the heart were harvested for were harvested for histochemical, biochemical, and molecular analysis to ascertain the effects of treatment on glycosphingolipid glycosyltransferase activity, gene/protein expression.We observed an age‐dependent increase in the left ventricular mass (LVmass) and decline in fractional shortening indicative of cardiac hypertrophy and dysfunction which worsens with western diet. This was accompanied by increased gene expression for atrial natriuretic protein, Brain‐type natriuretic protein, and increased protein expression of transforming growth factor, MAPK and AKT‐1. Masson’s trichrome staining revealed hypertrophy in cardiomyocytes and extensive fibrosis. In contrast, treatment with D‐PDMP preserved fractional shortening and the left ventricular mass to control levels over a 6 month treatment period. Also, the level of various cardiac glycosphingolipids and glycosyltransferases were decreased to control levels.Conclusion: Treatment with a glycosphingolipid glycosyltransferase inhibitor blunts cardiac hypertrophy in apoE‐/‐ mice fed a western diet.Grant Funding Source: Supported by NHLBI grant P01HL107153.
Site‐specific chemoenzymatic glycoengineering of cetuximab, a therapeutic monoclonal antibody (607.16)
Authors: Lomino Joseph , Giddens John , Wang Lai‐Xi 
Published: June 2021

IgG type monoclonal antibodies are an important class of therapeutic glycoproteins used in the treatment of a wide range of maladies: cancer, auto‐immune disorders, and infectious diseases. Cetuximab is a therapeutic monoclonal antibody, which is glycosylated at both the FAB and FC domains of the heavy chain at Asn ‐ 88 and Asn ‐ 299 respectively. The FAB glycan is composed of ca. 30% non‐human α ‐ gal and/or N‐glycolylneuramic acid (NGNA), which have been shown to cause allergic reactions in some patient populations, while the FC domain glycans are not optimal for effector functions. In this work, we perform site‐specific glycan remodeling of cetuximab by taking advantage of the substrate specificity of two endoglycosidases and their cognate glycosynthases. Endo F3, from E. meningosepticum, was initially used to remove glycans from both the FAB and FC domains. A fully galactosylated complex type glycan was precisely transferred to the FC using the glycosynthase mutant of Endo S from S. pyogenes. Finally a fully sialylated N‐glycan was added to the FAB glycosylation site using the Endo F3 glycosynthase mutant. The glyco‐reengineered cetuximab is expected to possess enhanced therapeutic efficacy without the allergic side effects. Our site‐specific glycosylation approach should be generally applicable to other therapeutic monoclonal antibodies.Grant Funding Source: NIH Grant R01 GM080374
Anti‐angiogenic and anti‐cancer property of a protein N‐glycosylation inhibitor (607.4)
Authors: Baksi Krishna , del Rosario Cristina , Banerjee Aditi , Zhang Zhenbo , Banerjee Dipak 
Published: June 2021

Tumor growth is angiogenesis dependent. Current therapeutics targets either tumor cells or microvasculature but not both. The patients therefore enjoy disease free survival only for a limited period of time, irrespective of a good initial prognosis and pathological response rates. Aberrant glycosylation has been observed in tumors and we have hypothesized that an asparagine‐linked (N‐linked) protein glycosylation inhibitor could target both endothelial cells and tumor cells and eliminate tumor progression. Our objective is to test the efficacy of Tunicamycin. The results support Tunicamycin treatment causes cell cycle arrest, develops endoplasmic reticulum (ER) stress and induces unfolded protein response (upr)‐mediated apoptosis. Analysis of C=O stretching vibrations of Amide I of proteome by Raman spectroscopy concluded protein denaturation. Tunicamycin treatment (i) prevented the colony formation, (ii) failed to reverse the inhibition of angiogenesis by VEGF165; and (iii) indiscriminately inhibited the progression of double‐ and a triple‐negative breast cancers by ~55‐65% in three weeks. Proliferation of MDA‐MB‐231, MDA‐MB‐468, BT‐20, BT‐474 and ZR‐731 human breast cancer cells was also inhibited. Induction of apoptosis in MDA‐MB‐23 confirmed translational attenuation of upr. Supported in part by grants from Susan G. Komen for the Cure BCTR0600582 (DKB) and NIH/NIMHD 2G12MD007583 (KB).Grant Funding Source: Supported in part by grants from Susan G. Komen for the Cure BCTR0600582 (DKB) and NIH/NIMHD 2G12MD0
CaM kinase I binds and regulates p53 in breast cancer cells (613.2)
Authors: Geck Renee , Schmitt John 
Published: June 2021

P53 is a tumor suppressor protein that regulates gene expression and cell cycle progression. Calcium/calmodulin‐dependent protein kinase (CaM kinase) proteins, including CaM KK and CaM KI, are known to control ERK and several transcription factors. CaM KI has also been shown to bind and phosphorylate the cell cycle protein p27 to promote cell cycle progression. Interestingly, p53 contains a possible CaM KI phosphorylation site at S366. Our goal was to evaluate the ability of CaM KI to bind and regulate p53 as well as investigate its effects on p53 phosphorylation, the cell cycle, and cell growth in MCF‐7 and MDA‐MB‐231 breast cancer cells. GST pull‐downs showed that CaM KI associates with phosphorylated p53 at S366. Treatment of MCF‐7 cells with estrogen (E2) increases the endogenous association of CaM KI‐p53 as does treatment of MDA cells with the calcium‐elevating agent, ionomycin. None of the treatments led to an association of p53 with CaM KK, CaM KII, or CaM KIV. Interestingly, E2 increased MCF‐7 progression through the cell cycle and MCF‐7 cell growth, effects enhanced by inhibiting p53 with PFT‐α. In summary, our data suggests that CaM KI binds and phosphorylates p53 in MCF‐7 and MDA cells, and that E2 and PFT‐α stimulate MCF‐7 cell growth through a novel CaM KI‐p53 pathway.Grant Funding Source: Supported by M.J.Murdock Charitable Trust #2011267
Profiling of human normal and cancer cell lines using phenotype microarray analysis (613.5)
Authors: Parmar Nitika , Wetton Nichole , Alvarado Shelley , Kennedy Shane 
Published: June 2021

The aim of this research project was to study how women’s cancer cell lines respond to anti‐cancer agents and whether the mTOR pathway was being targeted in them. Five cancer cell lines were used‐ two ovarian (OV90, TOV112D), two uterine (MES‐SA, KLE), one breast (SKBR) and one non‐cancerous kidney (HEK293). BIOLOG Phenotype Microarray anti‐cancer panels were used to profile these cell lines. Cells were seeded on a variety of BIOLOG panels consisting of 96 different anti‐cancer agents for 24‐36h. Cell growth was examined using a MTT cell proliferation assay. Six reagents‐ berberine chloride, azathioprine, celastrol, gossypol, miltefosine, and etoposide, had the largest growth impact on the cell lines. Cell lines were then treated with 5µM of these reagents and then subjected to the InstantOne Elisa assay to determine activation status of a protein‐pS6 kinase in the mTOR pathway. Cancer cells from different tissues responded differently to the same anti‐cancer agent; HEK293 cells showed maximum resistance to treatment with these compounds. The reagents were then used at doses varying from 0.05‐5μM to conduct a dose response profile. Growth inhibition was observed at a concentration of 0.5μM and above, in a cell line specific manner. Appropriate controls were used in all experiments. Our collective findings indicate that growth inhibition observed in some cell lines is due to the inhibition of the mTOR pathway.Grant Funding Source: Supported by the California State University Program for Education and Research in Biotechnology
Inhibition of ammonia oxidation in the thermophilic ammonia‐oxidizing archaeon Nitrosocaldus yellowstonii (617.6)
Authors: Gelsinger Diego , Cassani Madeline , Jewell Talia , de la Torre Jose 
Published: June 2021

Molecular data suggests ammonia‐oxidizing archaea (AOA) play an important role in nitrogen cycling. In particular, organisms related to the thermophilic archaeon Nitrosocaldus yellowstonii appear to be the only nitrifiers present in terrestrial hydrothermal systems at temperatures above 65°C. To better understand the contribution these thermophilic archaeal nitrifiers have in geochemical processes such as nitrogen cycling, we need tools to manipulate their metabolic activity in laboratory cultures and in situ. Nitrosocaldus yellowstonii HL72 is a thermophilic AOA cultivated from a hot spring in Yellowstone National Park (Wyoming, USA). To understand how N. yellowstonii contributes to nitrogen cycling in hot springs, we must first characterize the effect of nitrification inhibitors on its metabolic function. In this study, we have focused on allylthiourea (ATU), a commonly‐used inhibitor of nitrification. We have shown that ATU is stable at temperatures up to 72°C, the growth temperature of N. yellowstonii. Ammonia oxidation in N. yellowstonii is completely inhibited by ATU concentrations of 1 mM, concentrations that are approximately 100 times higher than those required to inhibit ammonia oxidation in bacteria. Addition of 1 mM ATU decreased the rate of nitrite production of N. yellowstonii from 0.039 ± 0.001 mM/h‐1 prior to addition, to 0.001 ± 0.001 mM/h‐1. Because nitrite is used for a proxy of growth, quantitative PCR was used to measure cell numbers for the single copy gene amoA per cell. There was no change in amoA copy number after the addition of 1 mM ATU while the unamended control continued to exponentially increase, further confirming that the addition of 1 mM ATU halted cell growth completely. RT‐qPCR showed amoA expression decreased 23.5% and 47.9% compared to the unamended control, two and four hours after ATU addition. We conclude that ATU is an effective inhibitor of AOA cell growth and is will be a useful tool to manipulate metabolic activity of AOA in hot spring systems.Grant Funding Source: Supported by NIH MARC T34‐GM00857