Oklahoma Medical Research Foundation Research Articles

Abstract 106: Diclofenac hybrids with oleanolic acid oximes modulate key signaling pathways in pancreatic cancer cells

Abstract Pancreatic cancer is characterized by constitutive activation of mitogen-activated protein kinases (MAPKs) like JNK and p38 determining its malignant phenotypes. Overexpression of NF-κB and COX-2 involved in inflammation contributes to increased cell growth and invasiveness. The latter might be related to the activated AKT pathway. Nonsteroidal anti-inflammatory drugs were suggested for pancreas cancers prophylaxis. Naturally occurring oleanolic acid (OA) was shown to possess anti-inflammatory and anti-tumorigenic activities. Coupling its analogs with NSAIDs may enhance this effect. Our study aimed to evaluate the effect of two oleanolic acid oximes derivatives conjugated with diclofenac (DCL), 4c (3-diclofenacoxyiminoolean-12-en-28-oic acid benzyl ester) and 4d (3-diclofenacoxyiminoolean-12-en-28-oic acid morpholide) on inflammation-related pathways, MAPK and NF-κB, cell cycle arrest and ROS formation in PSN-1 pancreatic cancer cells. PSN-1 cells were incubated with DCL, 4c or 4d at the concentration of 30µM. Activation of NF-κB was assessed by measuring binding of its p50 and p65 subunits to DNA using ELISA method. Their nuclear concentrations were determined by Western blot. RT-PCR was used to evaluate NF-κB subunits and COX-2 mRNA levels. Bead-based immunoassay determined the cytosolic levels of p38, JNK, CREB, AKT, P70S6K and STAT3 proteins. Cell cycle distribution and ROS formation were assessed using flow cytometry. Conjugate 4d significantly reduced nuclear level of p65 subunit and its mRNA expression, although compound 4c had more significant impact on COX-2 expression. Both derivatives significantly diminished p65 DNA binding, but only 4d had significant effect on p50. Both compounds reduced ROS production, but neither of them affected the cell cycle arrest. We observed decreased cytosolic AKT level along with increased levels of unphosphorylated JNK and p38 kinases which may have an antagonistic effect on proliferation by reducing activity of inflammatory mediators acting as tumor promoters. AKT decrease after treatment with 4d derivative might be the cause of increased unphosphorylated CREB level. Accumulation of p38 and JNK in cytosol could be a result of decreased ROS production and/or cause increased CREB protein level. Additionally, increased STAT3 cytosolic level suggests inhibition of mTORC1 pathway that explains reduced p70S6 kinase. Overall, these results suggest that OAO-DCL morpholide conjugate may affect both NF-κB and MAPK signaling pathways in pancreatic cancer cells. Further studies are required to explain detailed mechanism of these modifications. Funding: Studies were supported by Polish National Science Centre grant 2016/21/B/NZ7/01758. Acknowledgments: Authors thank Dr. Gary Gorbsky, Chair of Cell Cycle & Cancer Biology Program in Oklahoma Medical Research Foundation for supporting financing Maria Narozna conference participation. Citation Format: Maria Narozna, Violetta Krajka-Kuzniak, Robert Kleszcz, Wanda Baer-Dubowska. Diclofenac hybrids with oleanolic acid oximes modulate key signaling pathways in pancreatic cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 106.

Identification of Gene Networks Associated with the Anti-Leukemic Effect of Anti-Inflammatory Drugs on Acute Myeloid Leukemia Cell Lines

IntroductionDespite recent advances in therapy, acute myeloid leukemia (AML) remain a medical challenge with high morbidity and mortality rates. For most patients, allogeneic hematopoietic stem cell transplantation remain the only curative option, but due to the advanced age at diagnosis, a significant proportion of patients are not elegible to this form of therapy. Nevertheless, novel therapies are warranted. There is preclinical evidence that anti-inflammatory compounds, such as COX-2 inhibitors and steroids, may have anti-neoplastic activity in different tumor types, including AML; nevertheles the mechanisms associated with its anti-neoplastic activity are not clear. Therefore, the aim of this work was to evaluate the anti-leukemic effect of the anti-inflammatory compounds nimesulide and prednisolone in AML cell lines and to identify genes and molecular pathways associated with cytotoxicity through transcriptome analysis.MethodsThe leukemic cell lines HL-60, THP-1, OCI-AML2 and OCI-AML3 were treated with nimesulide and prednisolone at 100 µM alone and in combination and with cytarabine at 2.5 µM. Twenty four hours after treatment , we measured the amount of cell death using Annexin V Apoptosis Detection Kit FITC (ThermoFisher) and the cell cycle was analized after fixing the cells with 70% alcohol and incubation with propidium iodide (1mg/ml) and RNAse (10mg/ml). In another experiment, we harvested the cells after 4 hours of treatment for transcriptome analysis. RNA was extracted from control (DMSO) and treatment groups (1 - nimesulide, 2 - prednisolone , 3 - nimesulide and prednisolone) with RNeasy Mini Kit (Qiagen). The Illumina® NEBNext® Ultra II Directional RNA Library Prep Kit was used for library preparation, following the manufacturer protocol using Poly(A) mRNA Magnetic Isolation Module. Equimolar amount of libraries was sequenced using an Illumina NextSeq 500, following the manufacturer's instructions, on the Oklahoma Medical Research Foundation Genomics Core (USA). The sequences obtained with the RNA-Seq technique were aligned in the human genome of reference GRCh37.75 by the software Spliced Transcripts Alignment to a Reference (STAR) v2.5 and to obtain normalized counts in FPKM, the software Expectation-Maximization (RSEM) v1.3.0 was used. To identify network of genes correlated with the treatment (modules), we used the Weighted Correlation Network Analysis (WGCNA). Functional enrichment analysis of the WGCNA differentially expressed modules was performed using the Integrated Annotation, Visualization and Discovery Database (DAVID) v6.8 in order to correlate with biological processes.ResultsIn the cell cycle analysis, we observed a significant increase (p < 0.05) in the sub-G0 phase (cell death) after treatment with nimesulide alone, and in combination with prednisolone (figure 1). No effect was observed in the prednisolone only group. The cell cycle effect induced by nimesulide on HL-60 and OCI-AML2 was similar to the induced by cytarabine, a standard chemotherapy agent for AML that in known to induce arrest in the S phase. In addition, the cell line arrest in THP-1 was greater with nimesulide than with cytarabine, while OCI-AML3 was less sensitive to both nimesulide and cytarabine. Regarding cell death mechanism, treatment with nimesulide induced predominantly an increase in late apoptosis that was potentiated after combined treatment with nimesulide and cytarabine (figure 2). After the demonstration of cell cycle arrest and apoptosis induction after treatment with nimesulide, we performed whole transcriptome sequencing followed by WGCNA analysis. We have identified gene modules that were significantly correlated with anti-inflammatory treatments, being 1 module down-regulated (lightyellow with p = 0.00052) and 2 modules up-regulated (lightcyan with p = 0 .00025 and tan with p = 0.000038). Analysis of functional enrichment using DAVID showed up-regulation of gene networks associated with apoptotic processes and autophagy and down- regulation of gene networks associated with cell cycle and RNA splicing pathwaysConclusionsThe COX-2 inhibitor nimesulide caused cell cycle arrest and apoptosis in AML cell lines and potentiated the cytotoxic effects of cytarabine. This treatment was associated with up- regulation of autophagy and apoptosis and down-regulation of cell cycle and RNA splicing gene networks. [Display omitted] DisclosuresSantos: Novartis: Membership on an entity's Board of Directors or advisory committees, Research Funding; Amgen: Membership on an entity's Board of Directors or advisory committees; Abbvie: Membership on an entity's Board of Directors or advisory committees; Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees. Campregher: Astellas: Consultancy.

Editorial introductions

Editorial introductions

Systemic Lupus Erythematosus Research: A Bibliometric Analysis over a 50-Year Period.

Bibliometric analysis is a well-established approach to quantitatively assess scholarly productivity. However, there have been few assessments of research productivity on systemic lupus erythematosus (SLE) to date. The aim of this study was to analyze global research productivity through original articles published in journals indexed by the Web of Science from 1971 to 2020. Bibliometric data was obtained from the Science Citation Index Expanded in the Web of Science Core Collection database. Only original articles published between 1971 and 2020 on SLE were included in the analysis. Over the 50-year period, publication production in SLE research has steadily increased with a mean annual growth rate of 8.0%. A total of 44,967 articles published in 3435 different journals were identified. The journal Lupus published the largest number of articles (n = 3371; 8.0%). A total of 148 countries and regions contributed to the articles. The global productivity ranking was led by the United States (n = 11,244, 25.0%), followed by China (n = 4893, 10.9%). A three-field plot showed that the Oklahoma Medical Research Foundation and the Johns Hopkins University together contributed 18.5% of all articles from the United States. A co-occurrence network analysis revealed five highly connected clusters of SLE research. In conclusion, this bibliometric analysis provided a comprehensive overview of the status of SLE research, which could enable a better understanding of the development in this field in the past 50 years.

Change Is Inevitable

Change Is Inevitable

First person – Sarah Colijn

ABSTRACTFirst Person is a series of interviews with the first authors of a selection of papers published in Disease Models & Mechanisms (DMM), helping early-career researchers promote themselves alongside their papers. Sarah Colijn is first author on ‘Cell-specific and athero-protective roles for RIPK3 in a murine model of atherosclerosis’, published in DMM. Sarah conducted the research described in this article while a graduate student in Courtney Griffin's lab at Oklahoma Medical Research Foundation, Oklahoma City, OK, USA. She is now a Postdoctoral Research Associate in the lab of Amber Stratman at Washington University in St. Louis, School of Medicine, St. Louis, MO, USA, investigating how the cardiovascular system develops during embryogenesis in both physiological and pathological scenarios.

Abstract GMM-021: UTILIZATION OF PATIENT-DERIVED TUMOR XENOGRAFT (PDX) MODELS IN ONCOLOGY

Abstract BACKGROUND: The 5-year survival rate of patients with recurrent epithelial ovarian cancer is only 30%, in part due to relapse and resistance to first-line platinum-based chemotherapy. Even with marked progress toward understanding ovarian cancer biology, the translation of research findings into new therapies is still an enormous barrier to progress. Recent data suggests a 90% failure rate for new oncology drugs in the clinic. Development and preclinical testing of new cancer therapies has been limited by the availability of clinically relevant models that recapitulate the molecular and phenotypic characteristics of primary ovarian cancers. To overcome this barrier, we have initiated collaborative project between Stephenson Cancer Center (SCC) and Oklahoma Medical Research Foundation (OMRF) with the goal to generate a biobank of patient-derived xenograft (PDX) models derived directly from ovarian cancer patients undergoing therapy. RESULTS: PDX tumor models are generated in the Patient-Derived Xenograft and Preclinical Therapeutics (PDX-PCT) Core at OMRF and used to improve preclinical evaluation of new drugs towards more personalized medicine. We have collected blood samples and tumor tissues from consenting patients having primary debulking surgery at Stephenson Cancer Center since 2015 (&amp;gt;140 unique patients). Fresh tumor tissue is minced into small tumor chunks (~2 mm) and implanted into immunocompromised mice. PDXs are expanded for 3-4 passages and characterized based on tumor type, histology and molecular characteristics. Immunohistochemical and molecular analysis showed that histology of the original patient tumors as well as the expression of commonly used markers for high-grade serous ovarian cancer such as cytokeratin, PAX8 and WT1 are perfectly maintained in the derivative PDX lines. Genotyping of patients' tumors and their corresponding PDXs using Illumina's Exome Array revealed that single nucleotide polymorphism (SNP) profile found in the original tumors is perfectly maintained in PDX lines. One of the PDX limitations is that human tumor stromal and immune cells are being replaced with mouse equivalents upon in vivo propagation of PDXs, which negatively affects quality of genomic or proteomic analyses. To overcome this challenge, we optimized a method to deplete mouse cells from PDX tumors by magnetic cells separation. Our data showed that ovarian cancer PDXs contain around 28-54% of human tumor cells, where the rest are mouse cells. We also optimized protocols to utilize pure population of human tumor cells isolated from PDXs to transduce these with lentiviral vectors expressing GFP and/or luciferase to generate luciferized PDXs. Luciferized ovarian PDXs are excellent models used for non-invasive orthotopic tumor growth and drug response monitoring. CONCLUSIONS: Collection of our PDXs illustrates heterogeneity and diversity of human ovarian tumors. However, each PDX maintains essential molecular features of the original patients' tumor. Our goal is to generate clinically faithful ovarian tumor models that will provide a platform to learn more about cancer biology and to screen these for new better therapies. Citation Format: Magdalena Cybula, Katherine Moxley, Lin Wang, Luyao Wang, Magdalena Bieniasz. UTILIZATION OF PATIENT-DERIVED TUMOR XENOGRAFT (PDX) MODELS IN ONCOLOGY [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr GMM-021.

At BIO, business boomed, and drug pricing loomed

Some 17,000 people descended on Boston during the first week of June for the annual meeting of the Biotechnology Innovation Organization (BIO). If the crowds were any indication, business is booming. “This has been by far the most successful BIO convention we’ve ever had,” said John Maraganore, CEO of Alnylam Pharmaceuticals and chair of BIO. Attendees set a Guinness world record for the largest business-partnering event, with more than 45,000 meetings among people from across the globe. “If you want to be in the business of biotechnology, you need to be here,” said Manu Nair, who came to Boston from the Oklahoma Medical Research Foundation. Biotech investment is on track for a record year. “The first quarter was very strong for venture capital investing, and those trends seem durable,” said Glen Giovannetti, global biotechnology leader at the consulting firm EY. Cancer companies, especially immuno-oncology start-ups, are attracting a disproportionate amount

ATVB Named Lecture Review-Insight Into Author: James H. Morrissey.

HomeArteriosclerosis, Thrombosis, and Vascular BiologyVol. 37, No. 6ATVB Named Lecture Review—Insight Into Author: James H. Morrissey Free AccessNewsPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessNewsPDF/EPUBATVB Named Lecture Review—Insight Into Author: James H. Morrissey Originally published1 Jun 2017https://doi.org/10.1161/ATV.0000000000000055Arteriosclerosis, Thrombosis, and Vascular Biology. 2017;37:1041–1042ATVB Named Lecture Reviews–Sol Sherry Distinguished LectureInsight Into the Author: James H. Morrissey, PhD, University of Illinois at Urbana-ChampaignDownload figureDownload PowerPointWhy did you choose the profession of scientific investigation?I’ve always wanted to be a scientist, ever since I was a little child. Science fascinates me, and I couldn’t imagine doing anything different.Who have been your role model(s) in your scientific and professional life?I’ve had lots of good and bad examples of how to behave as a scientist, and I’ve learned a great deal from all of them!What have been important influences on your professional life?My wife, Genevieve, has given me the best guidance and grounding in my life. (We were high school sweethearts, so we’ve been together all our adult lives.) She’s helped me focus on my work while also becoming a more complete human being.What are your scientific inspirations?I became fascinated with biology when I was in high school. Biological systems are both incredibly complicated but also remarkably self-regulating, and it seemed to me that we understood very little of the mechanisms that made biological systems work. This made biology mysterious and exciting. I like focusing on hard problems, and I try to work on areas that aren’t the obvious next steps to take.How have mentors contributed to your professional development?Running a research lab is essentially running a small business, and you really have to spend a lot of time and energy supervising others. Scientists get no training for this, however! My best mentors have provided great examples of how to treat other people with kindness, dignity, and respect, while also being effective managers. Fletcher Taylor and Bill Thurman (formerly of the Oklahoma Medical Research Foundation) stand out in my memory.If you knew then what you know now, would you do anything different?I haven’t had a lot of regrets in my life. But I do wish I had learned to speak a foreign language well when I was young.What wisdom do you impart on new investigators?If you do get a position as a head of a laboratory, be sure to keep working in the lab for several years. At first, you will have the best hands of anyone in your lab, and the people you hire will need a lot of attention and help. Avoid the pressures that take you away from the lab bench for the first 5 years at least.If you were not a scientist, which profession would you pick?Computers, definitely.Which direction do you envisage your science taking?In recent years, I’ve found myself collaborating more and more intensely with others, especially with scientists who don’t work in my field but whose expertise is crucial to tackling a problem we are addressing. This has been highly rewarding to all concerned, and I see myself engaging in more extensive collaborations of this nature in the future.What are your nonscientific activities?Reading (especially, nineteenth-century literature), traveling, and riding my tandem bike with my wife.What sports do you follow?I have a passion for minor-league baseball. I try to attend a game whenever I visit a town that has minor league ball.What are your favorite books, movies, music (pick one or all)?One of my favorite books is also one of my favorite movies: To Kill a Mockingbird.What are you favorite foods and are they heart healthy?Well, that would be bacon. It isn’t heart healthy, so I only eat it on special occasions.FootnotesThis Insight Into ATVB Authors was originally published as part of the ATVB Named Lecture Review series. The original article is available online at http://atvb.ahajournals.org/content/35/6/1298.full Previous Back to top Next FiguresReferencesRelatedDetails June 2017Vol 37, Issue 6 Advertisement Article InformationMetrics © 2017 American Heart Association, Inc.https://doi.org/10.1161/ATV.0000000000000055PMID: 28539492 Originally publishedJune 1, 2017 PDF download Advertisement

Immediate Effect of a Service Dog on Walking Speed in Individuals with Multiple Sclerosis and Gait Dysfunction: A Pilot Study.

Walking impairment and physical inactivity are major concerns for individuals with multiple sclerosis (MS), with mobility being reported as the highest functional priority for 65% of patients with MS.1 Gait alterations can include slower speed, shorter stride length, and decreased distance walked, giving the individual the perception of decreased ability to perform activities of daily living.2 Service animals are increasingly being recognized as productive aids to individuals with a variety of neurologic disabilities. Service dogs are trained to do a range of tasks from retrieving objects to opening doors, pulling manual wheelchairs, and performing simple tasks at home that assist with activities of daily living. We designed a study to determine the immediate effect of the assistance of a service dog on walking speed in individuals with ambulatory dysfunction secondary to MS.The convenience sample comprised 44 individuals with identifiable gait abnormalities secondary to MS who were able to ambulate safely without the use of an assistive device even if they used one intermittently or on a regular basis to compensate for imbalance or motor deficits. Participants completed an informed consent form approved by the Oklahoma Medical Research Foundation institutional review board (Oklahoma City) before testing.Each individual performed two sets of Timed 25-Foot Walk testing (T25FW) at a fast but safe speed separated by 15 minutes of rest time. Each set involved one unassisted walk and one walk with the service dog, with 3 minutes of rest time in between. The second set was in the reverse order, walking with the service dog first. The dog was trained to walk beside the participant, so as not to pull or lag behind, and had a special harness with a flexible handle to eliminate the option of leaning on it for support. The usual dog handler walked on the opposite side to ensure compliance by the dog and to eliminate a potential influence from lack of experience with dog handling by the patient.The results of all descriptive analyses are reported as mean ± SE. A paired-samples t test was used to examine the difference between walking with and without a service dog. Statistical analysis was performed using IBM SPSS Statistics for Windows, version 20.0 (IBM Corp, Armonk, NY). Statistical significance was defined as P < .05.The demographic characteristics of the participants are presented in Table 1. There was a significant difference in mean ± SD T25FW times, with individuals walking faster with the aid of a service dog (8.78 ± 2.19 seconds) compared with walking independently (9.32 ± 2.69 seconds) (P = .003) (Figure 1).Impaired gait is a major problem for many individuals with MS because it limits their participation in regular activities, decreases their ability to participate in physical exercise, and increases their risk of injury and dependence on others. Limited mobility directly affects quality of life, social participation, and overall health, and it has been identified as the major determining factor predicting unemployment.3 Interventions to improve gait include ambulatory aids such as canes, walkers, ankle-foot orthoses, functional electrical stimulation, exercise, gait training, and medications.Previous research has shown that owning a dog can increase not only physical activity by walking4 but also social interaction5 and social acceptance, as well as inducing beneficial psychological and physiological changes.6 Direct health benefits include lower systolic blood pressure and cholesterol levels,7 higher self-esteem,8 increased exercise,7 and reduced anxiety and stress9 in adults and children.10 Having a trained service dog has also been shown to be economically beneficial, with one study showing a decrease in the number of paid assistance hours.11The present study is the first, to our knowledge, to evaluate the benefits of using a service animal on walking speed in MS. This study found an improvement in walking speed when individuals walked with the aid of a service dog compared with ambulating on their own. Walking speed has been used as an outcome in medical interventions directed at improving ambulation, including 4-aminopyridine (fampridine/dalfampridine) therapy. The demonstrated objective improvement in the T25FW time with the use of a service dog with the added potential benefits in physical and psychological well-being should encourage additional research in this area.The authors gratefully acknowledge Bamse, the service dog who lovingly and selflessly made this study possible.

J. Donald Capra, M.D. (1937–2015)

On February 24, 2015, at the age of 77, J. Donald “Don” Capra, M.D., president emeritus of the Oklahoma Medical Research Foundation, died of a malignant glioblastoma. For countless immunologists, Don was a pundit, enthusiast, and confidant. He was a member of The American Association of

A Novel C5 Complement Inhibitor Protects Against Sepsis-Induced Activation of Complement, Coagulation and Inflammation and Provides Survival Benefit in E. coli Sepsis

Bacterial sepsis triggers robust early activation of the complement system that leads to the formation of the C5b-9 terminal complex (TCC) and potent proinflammatory anaphylatoxins, C3a and C5a, which significantly contribute to organ failure and death. Complement inhibition at C3 level provides organ protection in low bacteremia experimental sepsis but may impair the clearance of the bacteria. Here we used a baboon model of E. coli sepsis and in vitro whole-blood assays to test the protective effects of a novel C5 inhibitor (RA101295, a 2036 Da cyclic peptide). In vitro whole blood assays were performed to study the effects of RA101295 on sepsis-induced complement activation, inflammation, oxidative stress and phagocytosis of the bacteria. Lepirudin-anticoagulated blood was incubated with 5x107 cfu/ml E. coli or 500 µg/ml lipopolysaccharide (LPS) for 30-240 min in the presence/absence of the inhibitor. Results showed that RA101295 strongly inhibited E. coli and LPS–dependent C5b-9 and C5a generation. Similarly, RA101295 inhibited the E. coliand LPS induced oxidative burst in neutrophils and monocytes by 40-50% with minimal decrease (<15%) on their phagocytic function.Next, we used a baboon model of E. coli sepsis to investigate the in vivo effects of RA101295. E. coli challenge (1-2x1010 cfu/kg; LD100) in baboons strongly induced the generation of soluble C5b-9 complexes. Treatment with RA101295 C5 inhibitor (four subcutaneous 10 mg/kg doses at 8-12 hrs intervals) fully inhibited complement activation. RA101295 inhibitor remarkably improved the clinical parameters of all treated animals. Two of three baboons survived the lethal dose; while the survival time of the third animal was increased by 300%. Animals treated with complement inhibitor displayed better heart and lung performance and had lower febrile response than non-treated septic baboons.RA101295 treatment of E. coli sepsis led to significantly lower consumptive coagulopathy as reflected by the APTT/PT, fibrinogen consumption and fibrin/fibrinogen degradation products. These data highlight the tight crosstalk between the coagulation and complement systems and suggest that the observed protective effects are due in part to dampening of sepsis-induced coagulopathy. The treatment during the bacteremic stage had no negative effects on bacterial clearance. Similar to the in vitro data, these results suggest that C5 inhibition with this peptide does not increase the risk of secondary infections and reoccurrence of sepsis, probably because is not interfering with the generation of C3b opsonin. Histologic analysis of organs confirmed that treatment with RA101295 provided substantial organ protection. In contrast to the non-treated group, RA101295 treated animals showed no obvious signs of thrombosis and capillary leak in the lungs; no follicular necrosis in the spleen and no tubular necrosis and glomerular thrombosis in the kidneys.Overall, our data demonstrate that RA101295 strongly inhibits LPS and E. coli induced complement activation and TCC without affecting the bacterial clearance. These results suggest that complement inhibition at C5 level represents a promising therapeutic target in the fight against sepsis-induced MOF. DisclosuresKeshari:Oklahoma Medical Research Foundation: Employment. Silasi-Mansat:Oklahoma Medical Research Foundation: Employment. Popescu:Oklahoma Medical Research Foundation: Employment. Chaaban:Oklahoma University Health Sciences Center: Employment. Lupu:Oklahoma Medical Research Foundation: Employment. DeMarco:Ra Pharmaceuticals: Employment. Lupu:American Heart Association: Membership on an entity’s Board of Directors or advisory committees; Merck: Research Funding; NIH: Research Funding; Oklahoma Medical Research Foundation: Employment.

Proceedings of the 2014 MidSouth Computational Biology and Bioinformatics Society (MCBIOS) Conference.

MidSouth Computational Biology and Bioinformatics Society (MCBIOS 2014) held its eleventh annual conference at the Wes Watkins Center at Oklahoma State University, Stillwater on March 7-8, 2014. theme was From Genome to Phenome: Connecting the Dots. Conference Chair this year was Rakesh Kaundal, who is also one of the MCBIOS board members, and conference committee members were Ulrich K. Melcher and Doris Kupfer. current president is Andy Perkins and Cesar Compadre was elected as President-Elect for 2015-16. There were 154 registrants and a total of 125 abstracts submitted (50 oral and 75 poster presentations). Keynote speakers were Owen White from University of Maryland School of Medicine, whose talk was titled The Human Microbiome Project: Large-Scale Data Management and Analysis, and Jody Hey from Temple University Designing Genealogy Samplers for Population Genetics. Dr. William Slikker, Director of the Food and Drug Administration's, National Center for Toxicological Research, concluded with a talk on the past ten years of MCBIOS and a perspective on its future. Participants also had the opportunity to attend a workshop on next-generation sequencing (NGS), hosted by Peter Hoyt of OSU and Dr. Graham Wiley of the Oklahoma Medical Research Foundation. workshop had a keynote by Dr. Joshua Orvis of University of Maryland School of Medicine and Johns Hopkins University on genome annotation. The winners of conference awards were: Best Paper Award: Zongliang Yue, Ping Wan, Hui Huang, Zhan Xie and Jake Y. Chen for SLDR: A Method to Identify New Gene Regulatory Relationship Candidates [1] Best Paper Runner-up: Nam S Vo and Vinhthuy Phan for Exploiting dependencies of pairwise-comparison outcomes to predict patterns of gene response [2] Best Oral Presentations (Post-Doctoral fellows): Michael A. Bauer, University of Arkansas for Medical Sciences Erich A. Peterson, University of Arkansas for Medical Sciences Best Oral Presentations (students): Karl Walker, Arkansas State University, 1st place Shraddha Thakkar, University of Arkansas for Medical Sciences, 2nd place Mihir Jaiswal, University of Arkansas at Little Rock, 3rd place Best Poster (Computation): Stephen Reichley, Mississippi State University, 1st place Kushal Bohra, Texas A&M University at Commerce, 2nd place Austin McCullough, John Brown University, 3rd place Best Poster (Biology): Shraddha Thakkar, University of Arkansas for Medical Sciences, 1st place Sunetra Das, University of Oklahoma, 2nd place (tie) Garima Saxena, University of North Texas, 2nd place (tie) Kangmei Zhao, University of Oklahoma, 2nd place (tie)

Petar Alaupovic

Petar Alaupovic, an internationally renowned biochemist and lipidologist, died on January 30, 2014, in Oklahoma City at the age of 91 years. Dr Alaupovic was a founder of the modern field of lipoproteins whose ideas continue to have an impact on its development. He performed research and taught at Oklahoma Medical Research Foundation since 1960. In 1964, Dr Alaupovic, known to his colleagues as Dr A or Pierre, proposed a revolutionary new classification system for lipoproteins. Until that time, the classification of lipoproteins focused on the physical properties of lipoproteins, emphasizing the lipid component and the size of the lipoprotein particle. However, Dr Alaupovic considered the protein part to have great biological importance that directed every step in the metabolism of lipoproteins and determined their effects on atherosclerosis. When he began his research on lipoproteins, there were considered to be just 2 apolipoproteins, apolipoprotein B and apolipoprotein A. Using immunologic and protein chemistry techniques, he isolated and characterized several additional apolipoproteins associated with lipoproteins. He proposed the ABC nomenclature for apolipoproteins, which initially included apolipoproteins A-I, A-II, B, C-I, C-II, C-III, D, and E. Most important to the subsequent progress in the field of lipoprotein, he subdivided the major lipoprotein classes, apolipoprotein A-I …

Systemic lupus erythematosus 2014.

Systemic lupus erythematosus 2014.

Editorial introductions

Editorial introductions

In Memoriam: Sven-Olof Olofsson (1947-2011)

In Memoriam: Sven-Olof Olofsson (1947-2011)

Heat Shock and Caloric Restriction Have a Synergistic Effect on the Heat Shock Response in a sir2.1-dependent Manner in Caenorhabditis elegans

The heat shock response (HSR) is responsible for maintaining cellular and organismal health through the regulation of proteostasis. Recent data demonstrating that the mammalian HSR is regulated by SIRT1 suggest that this response may be under metabolic control. To test this hypothesis, we have determined the effect of caloric restriction in Caenorhabditis elegans on activation of the HSR and have found a synergistic effect on the induction of hsp70 gene expression. The homolog of mammalian SIRT1 in C. elegans is Sir2.1. Using a mutated C. elegans strain with a sir2.1 deletion, we show that heat shock and caloric restriction cooperate to promote increased survivability and fitness in a sir2.1-dependent manner. Finally, we show that caloric restriction increases the ability of heat shock to preserve movement in a polyglutamine toxicity neurodegenerative disease model and that this effect is dependent on sir2.1.

Gene Expression of Defensin is Greatly Upregulated in Neonatal Neutrophils Following Labor (118.11)

Abstract This investigation documents the variability of gene expression between neonatal and adult neutrophils at rest and following stimulation with lipopolysaccharide. Our hypothesis is that distinct gene expression differences will be observed between neonatal and adult neutrophils at rest and following stimulation with LPS. Methods: Cord (9) and adult (12) blood samples were collected per routine procedures. Neutrophils were isolated and divided into 3 groups, which were seeded at 1 X 10 6 ml into multiwall plates and incubated with or without LPS 10 ng/µL (Invivogen) for 0, 45, and 90 minutes. RNA was isolated and gene expression differences determined via the Illumina platform at the Oklahoma Medical Research Foundation. Signal intensities from each probe were quantitated, adjusted for local background, and quantile normalized (Matlabs; Natwick, MA) to adjust the marginal distribution of each sample. Summary: Neonatal cord neutrophils were up-regulated in &amp;gt; 75% of differentially expressed genes at each time point as compared to adults. This difference remained relatively unchanged prior to and following LPS stimulation. Furthermore, neonatal neutrophils demonstrated a 13-16 fold increase in the expression of defensins prior to and following LPS stimulation. Conclusion: Hierarchical clustering of gene expression results showed distinct differences between adult and cord neutrophils in unstimulated and stimulated states.

Mapping the Intermedilysin-Human CD59 Receptor Interface Reveals a Deep Correspondence with the Binding Site on CD59 for Complement Binding Proteins C8α and C9

CD59 is a glycosylphosphatidylinositol-anchored protein that inhibits the assembly of the terminal complement membrane attack complex (MAC) pore, whereas Streptococcus intermedius intermedilysin (ILY), a pore forming cholesterol-dependent cytolysin (CDC), specifically binds to human CD59 (hCD59) to initiate the formation of its pore. The identification of the residues of ILY and hCD59 that form their binding interface revealed a remarkably deep correspondence between the hCD59 binding site for ILY and that for the MAC proteins C8α and C9. ILY disengages from hCD59 during the prepore to pore transition, suggesting that loss of this interaction is necessary to accommodate specific structural changes associated with this transition. Consistent with this scenario, mutants of hCD59 or ILY that increased the affinity of this interaction decreased the cytolytic activity by slowing the transition of the prepore to pore but not the assembly of the prepore oligomer. A signature motif was also identified in the hCD59 binding CDCs that revealed a new hCD59-binding member of the CDC family. Although the binding site on hCD59 for ILY, C8α, and C9 exhibits significant homology, no similarity exists in their binding sites for hCD59. Hence, ILY and the MAC proteins interact with common amino acids of hCD59 but lack detectable conservation in their binding sites for hCD59.