Biology Commons^™

The Engaged Microbiologist: Bringing The Microbiological Sciences To The K-12 Community, David J. Westenberg

Biological Sciences Faculty Research & Creative Works

Exposing K-12 students to cutting edge science that impacts their daily lives can bring classroom lessons to life. Citizen-science projects are an excellent way to bring high-level science to the classroom and help satisfy one of the cornerstone concepts of the Next Generation Science Standards (NGSS), "engaging in practices that scientists and engineers actually use." This can be a daunting task for teachers who may lack the background or resources to integrate these projects into the classroom. This is where scientific societies such as the American Society for Microbiology (ASM) can play a critical role. ASM encourages its members to …

Identification Of A Short Cell-Penetrating Peptide From Bovine Lactoferricin For Intracellular Delivery Of Dna In Human A549 Cells, Betty Revon Liu, Yue-Wern Huang, Robert Aronstam, Han-Jung Lee

Biological Sciences Faculty Research & Creative Works

Cell-penetrating peptides (CPPs) have been shown to deliver cargos, including protein, DNA, RNA, and nanomaterials, in fully active forms into live cells. Most of the CPP sequences in use today are based on non-native proteins that may be immunogenic. Here we demonstrate that the L5a CPP (RRWQW) from bovine lactoferricin (LFcin), stably and noncovalently complexed with plasmid DNA and prepared at an optimal nitrogen/phosphate ratio of 12, is able to efficiently enter into human lung cancer A549 cells. The L5a CPP delivered a plasmid containing the enhanced green fluorescent protein (EGFP) coding sequence that was subsequently expressed in …

Adropin Acts In Brain To Inhibit Water Drinking: Potential Interaction With The Orphan G Protein-Coupled Receptor, Gpr19, Lauren M. Stein, Gina L.C. Yosten, Willis K. Samson

Biological Sciences Faculty Research & Creative Works

Adropin, a recently described peptide hormone produced in the brain and liver, has been reported to have physiologically relevant actions on glucose homeostasis and lipogenesis, and to exert significant effect on endothelial function. We describe a central nervous system action of adropin to inhibit water drinking and identify a potential adropin receptor, the orphan G protein-coupled receptor, GPR19. Reduction in GPR19 mRNA levels in medial basal hypothalamus of male rats resulted in the loss of the inhibitory effect of adropin on water deprivation-induced thirst. The identification of a novel brain action of adropin and a candidate receptor for the peptide …

Aging-Related Impairment Of Urine-Concentrating Mechanisms Correlates With Dysregulation Of Adrenocortical Angiotensin Type 1 Receptors In Male Fischer Rats, Hong Ji, Wei Zheng, Xie Wu, Robert C. Speth, Joseph G. Verbalis, Lauren M. Stein, Gina L.C. Yosten, Willis K. Samson, Kathryn Sandberg

Biological Sciences Faculty Research & Creative Works

To investigate age-associated impairments in fluid homeostasis, 4-mo (young) and 32-mo (old) Fischer 344/BN male rats were studied before and after a dietary sodium load. Transferring young rats from a low-sodium (LS) to a high-sodium (HS) diet increased water intake and urine volume by 1.9-and 3.0-fold, respectively, while urine osmolality and plasma aldosterone decreased by 33 and 98%. Concomitantly, adrenocortical angiotensin type 1 receptor (AT1R) density decreased by 35%, and AT1bR mRNA decreased by 39%; no changes were observed in AT1aR mRNA. In contrast, the increase in water intake (1.4-fold) was lower in the old rats, and there was no …

Neuronostatin Acts Via Gpr107 To Increase Camp-Independent Pka Phosphorylation And Proglucagon Mrna Accumulation In Pancreatic Α-Cells, Mollisa M. Elrick, Willis K. Samson, John A. Corbett, Alison S. Salvatori, Lauren M. Stein, Grant R. Kolar, Aaron Naatz, Gina L.C. Yosten

Biological Sciences Faculty Research & Creative Works

Neuronostatin (NST) is a recently described peptide that is produced from the somatostatin preprohormone in pancreatic α-cells. NST has been shown to increase glucagon secretion from primary rat pancreatic islets in low-glucose conditions. Here, we demonstrate that NST increases proglucagon message in α-cells and identify a potential mechanism for NST's cellular activities, including the phosphorylation of PKA following activation of the G protein-coupled receptor, GPR107. GPR107 is abundantly expressed in the pancreas, particularly, in rodent and human α-cells. Compromise of GPR107 in pancreatic α-cells results in failure of NST to increase PKA phosphorylation and proglucagon mRNA levels. We also demonstrate …

Hypothalamic Action Of Phoenixin To Control Reproductive Hormone Secretion In Females: Importance Of The Orphan G Protein-Coupled Receptor Gpr173, Lauren M. Stein, Chloe W. Tullock, Stacy K. Mathews, David Garcia-Galiano, Carol F. Elias, Willis K. Samson, Gina L.C. Yosten

Biological Sciences Faculty Research & Creative Works

Sexual maturation and maintenance of reproductive function are regulated by neurohormonal communication between the hypothalamus, pituitary, and gonads (referred to as the HPG axis). Phoenixin (PNX) is a newly identified, endogenous peptide abundantly produced in the hypothalamus and shown to be an important mediator of ovarian cyclicity. However, the underlying mechanisms by which phoenixin functions within the HPG axis are unknown. Previous in vitro studies demonstrated a direct action of PNX on gonadotrophs to potentiate gonadotrophin-releasing hormone (GnRH) induced luteinizing hormone (LH) secretion. Therefore, we hypothesized that centrally derived phoenixin regulates the preovulatory LH surge required for ovarian cyclicity. We …

Phytoforensics Tools: The Degradation And Detection Of Chlorinated Solvents In Integrated Systems, Tommy J. Goodwin Jr.

Masters Theses

"Due to decades of mismanaged pollutants entering groundwater, subsurface pollution of various compounds has become a widespread challenge. Chlorinated solvents are the most common groundwater contaminants that persist in aquifers, and remediation of these wide-spread plumes is difficult. Bioremediation, permeable reactive barriers, and phytoremediation are remedial technologies that have been developed and applied to chlorinated solvents in groundwater systems. This study integrates these technologies in different combinations to demonstrate the remediation potential of this approach. Zerovalent iron (ZVI) and bioaugmentation with a Dehalococcoides sp. (DHC) culture were applied separately and in combination for degradation of perchloroethene (PCE). Salix pentandra were …

Correlation Between Sleep And Lifespan In Drosophila Melanogaster, Courtney H. Fiebelman

Masters Theses

”Sleep has previously been associated with lifespan. Monitoring sleep in any given fly over their lifetime facilitates the ability to predict the lifespan of that given fly. Using this estimate, lifespan can potentially correlate with biological age to identify when health parameters have declined.

To confirm that the prediction algorithm could identify short and long-lived flies, glutathione levels in heads and bodies were compared between two groups. The results showed this to be consistent in the bodies of wild-type Canton S male flies, and showed that glutathione was decreased in the predicted biologically older flies. These data show that glutathione …

Clusterin Seals The Ocular Surface Barrier In Mouse Dry Eye, Aditi Bauskar, Wendy J. Mack, Jerome Mauris, Pablo Argüeso, Martin Heur, Barbara A. Nagel, Grant R. Kolar, Martin E. Gleave, Takahiro Nakamura, Shigeru Kinoshita, Janet Moradian-Oldak, Noorjahan Panjwani, Stephen C. Pflugfelder, Mark R. Wilson

Biological Sciences Faculty Research & Creative Works

Dry eye is a common disorder caused by inadequate hydration of the ocular surface that results in disruption of barrier function. The homeostatic protein clusterin (CLU) is prominent at fluid-tissue interfaces throughout the body. CLU levels are reduced at the ocular surface in human inflammatory disorders that manifest as severe dry eye, as well as in a preclinical mouse model for desiccating stress that mimics dry eye. Using this mouse model, we show here that CLU prevents and ameliorates ocular surface barrier disruption by a remarkable sealing mechanism dependent on attainment of a critical all-or-none concentration. When the CLU level …

Constitutive Activity Among Orphan Class-A G Protein Coupled Receptors, Adam L. Martin, Michael A. Steurer, Robert Aronstam

Biological Sciences Faculty Research & Creative Works

The purpose of this study was to evaluate the extent of constitutive activity among orphan class-A G protein coupled receptors within the cAMP signaling pathway. Constitutive signaling was revealed by changes in gene expression under control of the cAMP response element. Gene expression was measured in Chinese hamster ovary cells transiently cotransfected with plasmids containing a luciferase reporter and orphan receptor. Criteria adopted for defining constitutive activation were: 1) 200% elevation over baseline reporter gene expression; 2) 40% inhibition of baseline expression; and 3) 40% inhibition of expression stimulated by 3 μM forskolin. Five patterns of activity were noted: 1) …

Delivery Of Nucleic Acids And Nanomaterials By Cell-Penetrating Peptides: Opportunities And Challenges, Yue-Wern Huang, Han-Jung Lee, Larry Tolliver, Robert Aronstam

Biological Sciences Faculty Research & Creative Works

Many viral and nonviral systems have been developed to aid delivery of biologically active molecules into cells. Among these, cell-penetrating peptides (CPPs) have received increasing attention in the past two decades for biomedical applications. In this review, we focus on opportunities and challenges associated with CPP delivery of nucleic acids and nanomaterials. We first describe the nature of versatile CPPs and their interactions with various types of cargoes. We then discuss in vivo and in vitro delivery of nucleic acids and nanomaterials by CPPs. Studies on the mechanisms of cellular entry and limitations in the methods used are detailed.

The Physiology Of Proinsulin C-Peptide: Unanswered Questions And A Proposed Model, Gina L.C. Yosten, Grant R. Kolar

Biological Sciences Faculty Research & Creative Works

C-peptide is produced, processed, and secreted with insulin, and appears to exert separate but intimately related effects. In this review, we address the existence of the C-peptide receptor, the interaction between C-peptide and insulin, and the potential physiological significance of proinsulin C-peptide. C-peptide is produced, processed, and secreted with insulin, and appears to exert separate but intimately related effects. In this review, we address the existence of the C-peptide receptor, the interaction between C-peptide and insulin, and the potential physiological significance of proinsulin C-peptide.

Microbial Diversity: A Journey Through Carl Woese's Tree Of Life, David J. Westenberg

Biological Sciences Faculty Research & Creative Works

Review of: Microbial Diversity; James W. Brown; (2015). American Society for Microbiology Press, Washington, DC. 416 pages.

The Lxr Inverse Agonist Sr9238 Suppresses Fibrosis In A Model Of Non-Alcoholic Steatohepatitis, Kristine Griffett, Ryan D. Welch, Colin A. Flaveny, Grant R. Kolar, Brent A. Neuschwander-Tetri, Thomas P. Burris

Biological Sciences Faculty Research & Creative Works

Objective: Non-alcoholic steatohepatitis (NASH) is characterized by hepatic steatosis, inflammation and fibrosis. There are currently no targeted therapies for NASH. We developed a liver-specific LXR inverse agonist, SR9238, which effectively reduces hepatic lipogenesis in models of obesity and hepatic steatosis. We hypothesized that suppression of lipogenesis, which is pathologically elevated in NASH may suppress progression of hepatic steatosis to NASH. Methods: NASH was induced in B6 V-lep ^ob/J (ob/ob) mice using a custom complete rodent diet (HTF) containing high amounts of trans-fat, fructose, and cholesterol. Once NASH was induced, mice were treated with SR9238 for one month by i.p. …

The Loss Of Hh Responsiveness By A Non-Ciliary Gli2 Variant, Jinling Liu, Huiqing Zeng, Aimin Liu

Engineering Management and Systems Engineering Faculty Research & Creative Works

Hedgehog signaling is crucial for vertebrate development and physiology. Gli2, the primary effector of Hedgehog signaling, localizes to the tip of the primary cilium, but the importance of its ciliary localization remains unclear. We address the roles of Gli2 ciliary localization by replacing endogenous Gli2 with Gli2^ΔCLR, a Gli2 variant not localizing to the cilium. The resulting Gli2^ΔCLRKI and Gli2^ΔCLRKI;Gli3 double mutants resemble Gli2-null and Gli2;Gli3 double mutants, respectively, suggesting the lack of Gli2^ΔCLR activation in development. Significantly, Gli2^ΔCLR cannot be activated either by pharmacochemical activation of Smo in vitro or by loss …

Dephosphorylation Of Iqg1 By Cdc14 Regulates Cytokinesis In Budding Yeast, Daniel P. Miller, Hana Kenton Hall, Ryan Chaparian, Madison Mara, Alison Mueller, Mark C. Hall, Katie Shannon

Biological Sciences Faculty Research & Creative Works

Cytokinesis separates cells by contraction of a ring composed of filamentous actin (F-actin) and type II myosin. Iqg1, an IQGAP family member, is an essential protein in Saccharomyces cerevisiae required for assembly and contraction of the actomyosin ring. Localization of F-actin to the ring occurs only after anaphase and is mediated by the calponin homology domain (CHD) of Iqg1, but the regulatory mechanisms that temporally restrict actin ring assembly are not well defined. We tested the hypothesis that dephosphorylation of four perfect cyclin-dependent kinase (Cdk) sites flanking the CHD promotes actin ring formation, using site-specific alanine mutants. Cells expressing the …

Excessive Daytime Sleepiness Is Associated With Changes In Salivary Inflammatory Genes Transcripts, Matthew S. Thimgan, Cristina Toedebusch, Jennifer Mcleland, Stephen P. Duntley, Paul J. Shaw

Biological Sciences Faculty Research & Creative Works

Excessive daytime sleepiness (EDS) is a ubiquitous problem that affects public health and safety. A test that can reliably identify individuals that suffer from EDS is needed. In contrast to other methods, salivary biomarkers are an objective, inexpensive, and noninvasive method to identify individuals with inadequate sleep. Although we have previously shown that inflammatory genes are elevated in saliva samples taken from sleep deprived individuals, it is unclear if inflammatory genes will be elevated in clinical populations with EDS. In this study, salivary samples from individuals with sleep apnea were evaluated using the Taqman low density inflammation array. Transcript levels …

Increased Susceptibility To Bladder Inflammation In Smokers: Targeting The Paf–Paf Receptor Interaction To Manage Inflammatory Cell Recruitment, John Marentette, Grant Kolar, Jane Mchowat

Biological Sciences Faculty Research & Creative Works

Chronic bladder inflammation can result in a significant reduction in quality of life. Smoking remains a leading preventable risk factor in many diseases. Despite the large amount of evidence supporting the risks of smoking, roughly 45 million people in the United States remain smokers. The impact of cigarette smoking on inflammation is well established, but how smoking promotes bladder inflammation is currently unknown. The aim of this study was to determine if cigarette smoke exposure impacts inflammatory cell adherence to bladder endothelial cells and if targeting the platelet-activating factor (PAF)– PAF receptor (PAFR) interaction could be beneficial in managing bladder …

Increased Susceptibility To Bladder Inflammation In Smokers: Targeting The Paf–Paf Receptor Interaction To Manage Inflammatory Cell Recruitment, John Marentette, Grant Kolar, Jane Mchowat

Biological Sciences Faculty Research & Creative Works

Chronic bladder inflammation can result in a significant reduction in quality of life. Smoking remains a leading preventable risk factor in many diseases. Despite the large amount of evidence supporting the risks of smoking, roughly 45 million people in the United States remain smokers. The impact of cigarette smoking on inflammation is well established, but how smoking promotes bladder inflammation is currently unknown. The aim of this study was to determine if cigarette smoke exposure impacts inflammatory cell adherence to bladder endothelial cells and if targeting the platelet-activating factor (PAF)– PAF receptor (PAFR) interaction could be beneficial in managing bladder …

2015 Annual Report Department Of Biological Sciences, Ronald L. Frank, Chen Hou, Yue-Wern Huang, Adam R. Martin, Melanie R. Mormile, Dev Niyogi, Julie A. Semon, Katie S. Shannon, Matthew S. Thimgan, David J. Westenberg, Terry Wilson

Biological Sciences Faculty Research & Creative Works

No abstract provided.

Angiogenic Effect Of Bioactive Borate Glass Microfibers And Beads In The Hairless Mouse, Richard J. Watters, Roger F. Brown, D. E. Day

Biological Sciences Faculty Research & Creative Works

The purpose of this project was to investigate the angiogenic mechanism of bioactive borate glass for soft tissue repair in a 'hairless' SKH1 mouse model. Subcutaneous microvascular responses to bioactive glass microfibers (45S5, 13-93B3, and 13-93B3Cu) and bioactive glass beads (13-93, 13-93B3, and 13-93B3Cu) were assessed via: noninvasive imaging of skin microvasculature; histomorphometry of microvascular densities; and quantitative PCR measurements of mRNA expression of VEGF and FGF-2 cytokines. Live imaging via dorsal skin windows showed the formation at two weeks of a halo-like structure infused with microvessels surrounding implanted borate-based 13-93B3 and 13-93B3Cu glass beads, a response not observed with …

Sensitivity Of Cellular Oxidative Damage To Biosynthetic Rate And Metabolic Rate, Palihawadana Walauwe Kamalika Kaushalya Amunugama

Masters Theses

"The relationship between energy expenditure and longevity is a long standing question in aging studies. The empirical results have contradictory effects on the existing theories. A theoretical model and an experimental test of it were presented, revealing the detailed tradeoffs between metabolic rate, biosynthetic rate and cellular damage level. The dissipative mechanisms of oxidative metabolism cause various forms of cellular damages. To counteract the accumulation of damage, organisms have evolved highly efficient maintenance mechanisms. If there is no other energy demand possess, then most of the cellular damage would be repaired, regardless of how metabolic rate varies. However, the repairing …

Healing Of Bone Defects In A Rodent Calvarial Defect Model Using Strong Porous Bioactive Glass (13-93) Scaffolds, Yinan Lin

Masters Theses

"The main objective of this project was to evaluate the capacity of strong porous silicate (13-93) bioactive glass scaffolds prepared by a robocasting technique to regenerate bone and stimulate angiogenesis in a rat calvarial defect model. The scaffolds were created with the same grid-like microstructure but in a variety of formulations: (i) as-fabricated, (ii) pretreated in an aqueous phosphate solution to convert the glass surface to hydroxyapatite, (iii) loaded with bone morphogenetic protein-2 (BMP2) (1 µg per scaffold), and (iv) doped with copper (0.4-2.0 wt. % CuO). When compared to the as-fabricated scaffolds, the pretreated scaffolds enhanced bone regeneration at …

Ph Dependent Antibiotic Resistance Of An Alkaliphilic, Halotolerant Bacterium Isolated From Soap Lake, Washington, Tiffany Charlynn Edwards

Masters Theses

"Soap Lake, located in Washington State, is a meromictic, soda lake. Many bacterial isolates retrieved from Soap Lake have been noted to possess resistance to multiple antibiotics. A likely explanation for the wide range of antibacterial resistance exhibited by these strains is due to the impact of high alkalinity on the antibiotics themselves and not due to the presence of antibiotic resistance genes. The aim of our study was to determine if select antibiotics are effective against Halomonas eurialkalitoleranis, a bacterium capable of growth over a wide range of neutral to alkaline pH values, to investigate the influence of alkalinity …

Constitutive Activity In Orphan G Protein Coupled Receptors, Adam Lee Martin

Doctoral Dissertations

"The goal of this research was to use a distal signaling pathway analysis to evaluate the extent of agonist independent constitutive signaling among orphan class-A G protein coupled receptors (GPCRs). These receptors translate extracellular signals via conformational change into intracellular activation of different G proteins and subsequent second messenger synthesis. These small molecules regulate cellular biochemistry, eventually leading to nuclear signaling that results in changes in gene expression. Some GPCRs are capable of signaling in the absence of an activating ligand, a phenomenon called constitutive activity that is inhibited via an "inverse-agonist". The use of cAMP dependent Luciferase expression is …

Physiological Effects And Therapeutic Potential Of Proinsulin C-Peptide, Gina L.C. Yosten, Christine Maric-Bilkan, Patrizia Luppi, John Wahren

Biological Sciences Faculty Research & Creative Works

Connecting Peptide, or C-peptide, is a product of the insulin prohormone, and is released with and in amounts equimolar to those of insulin. While it was once thought that C-peptide was biologically inert and had little biological significance beyond its role in the proper folding of insulin, it is now known that C-peptide binds specifically to the cell membranes of a variety of tissues and initiates specific intracellular signaling cascades that are pertussis toxin sensitive. Although it is now clear that C-peptide is a biologically active molecule, controversy still remains as to the physiological significance of the peptide. Interestingly, C-peptide …

Low O2-Induced Atp Release From Erythrocytes Of Humans With Type 2 Diabetes Is Restored By Physiological Ratios Of C-Peptide And Insulin, Jennifer P. Richards, Gina L.C. Yosten, Grant R. Kolar, Cory W. Jones, Alan H. Stephenson, Mary L. Ellsworth, Randy S. Sprague

Biological Sciences Faculty Research & Creative Works

ATP release from erythrocytes in response to reduced oxygen (O2) tension stimulates local vasodilation, enabling these cells to direct perfusion to areas in skeletal muscle in need of O2. Erythrocytes of humans with type 2 diabetes do not release ATP in response to low O2. Both C-peptide and insulin individually inhibit low O2-induced ATP release from healthy human erythrocytes, yet when co administered at physiological concentrations and ratios, no inhibition is seen. Here, we determined: that 1) eryth-rocytes of healthy humans and humans with type 2 diabetes possess a C-peptide receptor (GPR146), 2) the combination of C-peptide and insulin at …

Neuropathology And Genetics Of Cerebroretinal Vasculopathies, Grant R. Kolar, Parul H. Kothari, Negar Khanlou, Joanna C. Jen, Robert E. Schmidt, Harry V. Vinters

Biological Sciences Faculty Research & Creative Works

Cerebroretinal vasculopathy (CRV) and the related diseases hereditary endotheliopathy with retinopathy, neuropathy, and stroke (HERNS), hereditary vascular retinopathy (HVR) and hereditary systemic angiopathy (HSA) [subsequently combined as retinovasculopathy and cerebral leukodystrophy (RVCL)] are devastating autosomal-dominant disorders of early to middle-age onset presenting with a core constellation of neurologic and ophthalmologic findings. This family of diseases is linked by specific mutations targeting a core region of a gene. Frameshift mutations in the carboxyl-terminus of three prime exonuclease-1 (TREX1), the major mammalian 3' to 5' DNA exonuclease on chromosome 3p21.1-p21.3, result in a systemic vasculopathy that follows an approximately 5-year course leading …

Sex And Basic Science. A Title Ix Position, Kathryn Sandberg, Joseph G. Verbalis, Gina L.C. Yosten, Willis K. Samson

Biological Sciences Faculty Research & Creative Works

No abstract provided.

Rapidly Self-Renewing Human Multipotent Marrow Stromal Cells (Hmsc) Express Sialyl Lewis X And Actively Adhere To Arterial Endothelium In A Chick Embryo Model System, Harris E. Mcferrin, Scott D. Olson, Miriam V. Gutschow, Julie A. Semon, Deborah E. Sullivan, Darwin J. Prockop

Biological Sciences Faculty Research & Creative Works

Background: There have been conflicting observations regarding the receptors utilized by human multipotent mesenchymal bone marrow stromal cells (hMSC) to adhere to endothelial cells (EC). To address the discrepancies, we performed experiments with cells prepared with a standardized, low-density protocol preserving a sub-population of small cells that are rapidly self-renewing.

Methods: Sialyl Lewis X (SLeX) and α4 integrin expression were determined by flow cytometry. Fucosyltransferase expression was determined by quantitative realtime RT-PCR. Cell adhesion assays were carried out with a panel of endothelial cells from arteries, veins and the microvasculature in vitro. In Vivo experiments were performed to determine …