Members

Former Lab Members:

  • Van Vo, Ph.D., Postdoctoral Research Assoc. Project: PKG1alpha (PKG-I-alpha) kinase/catalytic activity in PANCREATIC BETA-CELLS identified as a key enhancer of insulin signaling (e.g. enhancing Akt phosphorylation & activation, increasing FoxO1 phosphorylation & suppression of nuclear accumulation, & downstream increased in the expression of PDX-1), all promoting increases in beta-cell SURVIVAL & PROLIFERATION, thus promoting beta-cells REGENERATION & IMPROVING INSULIN SECRETION to prevent Type-1 & Type-2 DIABETES. (see our Publications, Wong JC, Vo V, Gorjala P & RR Fiscus, 2017)

Van also studied the cytoprotective & ANTI-DIABETES role of PDE-9 inhibitors (e.g. BAY 73-6691) & PDE-5 inhibitors (e.g. Sildenafil / VIAGRA) on pancreatic beta-cell survival & regeneration as potential new repurposing of these agents for preventing and treating Type-1 & Type-2 DIABETES.

  • Steve Morrill, M.S. graduate student at the University of Arizona, Tucson, AZ , with all research being conducted in the R. R. Fiscus Lab at Roseman University of Health Sciences, College of Medicine at the Summerlin Campus, Summerlin-Las Vegas, NV 89135. Steve also works as a full-time high school teacher in Molecular Biology at Faith Lutheran High School, Summerlin-Las Vegas, NV. Research Project: Co-localization of PKG1alpha (PKG-I-alpha) with amyloid precursor protein (APP) in cholinergic NG108-15 cells (used as a cell culture model of Alzheimer's disease) & the effects of PDE-9 inhibitor BAY 73-6691 on PKG1alpha-mediated phosphorylation of CREB & expression levels of APP.

Resulting M.S. Thesis (available ONLINE from the University of Arizona library): Effects of phosphodiesterase inhibitor BAY 73-6691 on expression/phosphorylation of CREB & expression of amyloid precursor protein (APP) in NG108-15 neuronal cells (Model of Alzheimer's disease). The NG108-15 cells possess the biochemical machinery for APP synthesis & processing, including release of the Alzheimer's-associated peptide A-beta that forms Amyloid Plaques.

The RR Fiscus lab has previously shown that NG108-15 cells express exclusively the PKG-I-alpha splice variant of PKG (assessed via ultra-sensitive cIEF Immunoassay with NanoPro1000, see Fiscus & Johlfs, 2012, Book Chapter in NEUROMETHODS: "Protein Kinase G (PKG): Involvement in Promoting Neural Cell Survival, Proliferation, Synptogenesis and Synaptic Plasticity and the Use of New Ultrasensitive Capillary-Electrophoresis-based Methodologies for Measuring PKG Expression and Molecular Actions") & that the PKG-I-alpha isoform is required for neuronal cell survival, likely involving phosphorylation of CREB & other downstream target proteins of the PKG-I-alpha SPLICE VARIANT, such as Akt, BAD, GSK-3-beta, RhoA, c-Src & VASP.

This study follows up of previous findings from our lab (Barger, Fiscus, Ruth, Hofmann & Mattson, 1995; and our many other publications over the last 25 years showing neuroprotection of neuronal cells by exposure to healthy-level NO (0.01 - 1 nanomolar/nM) as well as NATRIURETIC PEPTIDES (ANP & BNP at 1 - 100 nM), all via activation of PKG catalytic/kinase activity). Other labs have confirmed the special neuroprotective role of cerebrovascular eNOS (PREVENTING ALZHEIMER'S DISEASE), healthy-level NO & neural cell cGMP. For example, the healthy/physiological-level release of NO from eNOS in cerebrovascular endothelial cells plays an essential role in protecting nearby neurons in the memory-storing regions of the brain and specifically protecting the neuron's memory-storing synaptic connections (Susan A. Austin et al., in Katusic Lab at Mayo Clinic, 2013; Katusic & Austin, 2014). Others have shown that physiological-level NO, via elevation of cGMP & enhancing PKG activity, alters the processing of APP in neurons by up-regulating activity of alpha-secretase and down-regulating activity of beta-secretase (BACE1), resulting in decreased production of the toxic amyloid-beta peptide (A-beta) and increased production & secretion of the alternate APP product, the neuroprotective/synapto-protective/memory-protective sAPP-alpha (Kwak YD et al., 2011).

Steve Morrill's data from the R.R. Fiscus Lab, showing PKG-I-alpha co-localization with APP (on the microtubules throughout neuronal cells), now sheds further light about the mechanism of NO, natriuretic peptides (ANP, BNP, & CNP), Insulin, & sAPP-alpha (endogenous factors that all stimulate cGMP biosynthesis & cellular accumulation, causing downstream stimulation of PKG-I-alpha kinase activity) in regulating APP processing in neurons and the downstream synapto-protective effects. The data provide valuable new evidence for the role of PKG-I-alpha activation in preventing neurodegenerative disease like Alzheimer's disease and suggesting the use of PKG-I-alpha stimulators, such as PDE 1, 2, 5 & 9 inhibitors as new PHARMACEUTICAL AGENTS for treating Alzheimer's & other neurodegenerative diseases.

  • Mary G. Johlfs, B.S., M.S., Previously a Senior Research Associate and the Lab Manager of the Fiscus Lab at the Nevada Cancer Institute. Currently, she serves as the Director of Research Operations and Scientist for Roseman University of Health Sciences, Summerlin Campus. In that role, Mary helped Dr. Fiscus to set up almost all of the current state-of-the-art research instrumentation, including: 1) the Capillary Isoelectic Focusing (cIEF, NanoPro 100 and NanoPro 1000) instrumentation (used for advanced proteomic analysis for ultra-sensitive assessment of protein expression & post-translational modification, possessing 1,000-times higher sensitivity compared with conventional Western blots), 2) the Nikon confocal & TIRF (Total Internal Reflection Fluorescence) microscopes, 3) the Sony FASC (fluorescence-activated cell sorter), and 4) the Li-Cor NIRF (Near InfraRed Fluorescence) imager & quantification systems, located in our shared-equipment labs at Roseman's Summerlin campus.

Mary, as Director of Research Operataion for Roseman University's new College of Medicine in Summerlin-Las Vegas, NV, has also collaborated with the R.R. Fiscus Lab. Collaborative Project: Devlopment of new applications for the ultra-sensitive cIEF NanoPro 1000 proteomics instrument for indentifying NOVEL BIOMARKERS for Alzheimer's, Parkinson's and Lewy Body Dementia (LBD), for differential and early diagnosis via appropriate biomarkers of Protein Splice Variants & Post-translational modifications (PTMs). (see our publication, Johlfs M.G. et al. 2015; and the above mentioned 2012 Book Chapter in "Protein KInase Technologies-Neuromethods" author by Fiscus, R.R. & Johlfs, M.G. based on data showing the novel technical application of cIEF-NanoPro1000 for Neurological Diseases.

  • Janica C. Wong, Ph.D., Previously a Ph.D. graduate student in the Biochemistry program of the Department of Chemistry, University of Nevada Las Vegas, with all of her research being conducted in the Fiscus Lab, first at the Nevada Cancer Institute and then at Roseman University of Health Sciences. Janica has now published 6 peer-reviewed scienctific articles and 3 book chapters based on her research in the Fiscus Lab (see Publications). After leaving the Fiscus Lab, Janica continued in medical research, working as a Postdoctoral Fellow in Pediatric Endocrinology at Stanford University School of Medicine for 2 & 1/2 years. Janica is currently a Scientist at Merck & Co. in the San Francisco Bay Area.

  • Ben Costantino, Ph.D. Previously a Postdoctoral Research Associate in the Fiscus Lab at Roseman University. Ben was a key contributor to the research that lead to our new ultra-sensitive, non-radioactive, near-infrared-fluorescence (NIRF)-based methodology for quantification of kinase catalytic activity of any protein kinase, and the resulting Patent application (see Home page). Ben is currently working as a Scientist at Ventana Diagnostics (Roche), Tucson, AZ.

  • Priyatham Gorjala, Ph.D. Previously a Postdoctoral Research Associate in the Fiscus Lab at Roseman University, contributing to 2 peer-reviewed scientific articles and a book chapter (see Publications). Priyatham is currently working in the lab of Dr. Ranjana Mitra (former member of the lab of Dr. Oscar B. Goodman, Jr.), College of Medicine, Roseman Univeristy of Health Sciences.

  • Madhavi Bathina, M.S. Previously a Research Assistant II in the Fiscus Lab at the Nevada Cancer Institute, Summerlin-Las Vegas, NV. Madhavi contributed to one of our key publications, showing that PKG-I-alpha is hyper-activated in non-small cell lung cancer (NSCLC) cells, which catalyzes the major part of the phosphorylation of CREB (a pro-growth & pro-survival transcription factor), resulting in exaggerated expression of the cell-survival proteins Mcl-1, cIAP-1, Livin & Survivin, and the exaggerated proliferation and resistance to chemotherapy (chemoresistance) in these NSCLC cells. Madhavi left the Fiscus Lab to continue her research at St. Jude Children Research Hospital, followed by the University of Arizona-Tucson and currently City of Hope Comprehensive Cancer Center.

Research Collaborators:

  • Mary G. Johlfs, former member of the Fiscus Lab (see above) and currently the Director of Research Operation and Scientist at the Summerlin Campus of Roseman University. Mary oversees the operation of all of the advanced instrumentation at our Summerlin Campus, including the cIEF-based NanoPro 1000 proteomics system, the Nikon confocal/TIRF microscopes, the Sony FACS and the near infrared fluorescence imaging system (LI-COR). Mary continues her medical research working in collaboration with the Fiscus Lab. The collaborative project identifies novel biomarkers for the early diagnosis (and differentiation) of Alzheimer's, Parkinson's and Lewy Body Dementia, using Mary's special expertise with the ultra-sensitive NanoPro 1000 advanced proteomics system. Mary's previous publications, including peer-reviewed scientific articles and invited book chapters, have shown that the NanoPro 1000 system is far superior to conventional technologies for protein analysis, including having 1,000-times higher sensitivity compared with conventional Western blots and 100,000-times higher sensitivity compared with 2-D gels/Mass Spec (Johlfs et al., 2015, see Publications).

Because of this exquisitely-high sensitivity, the NanoPro 1000 eIEF instrumentation is able to analyze proteins and protein post-translational modifications in extremely small sample sizes of biological materials (biopsy, blood, cerebrospinal fluid, urine, saliva, etc.) for the analysis of biomarkers (see Publications). Also, the NanoPro 1000 system has clear advantages over ELISAs, that is commonly used for analysis of biomarkers, because the cIEF technology is able to identify off-target binding of the antibodies (because proteins are separated by high-resolution isoelectric focusing before recognition by the antibodies). Thus, the contribution of off-target binding, which is a common problem with ELISAs, can be eliminated, allowing accurate identification of the biomarkers. The NanoPro 1000 system is ideal for the discovery of novel biomarkers, such as unique/pathologically-associated isoforms of proteins and their post-translational modifications.

  • Tim Thuc Le, Ph.D., Assistant Professor, & Yasuyo Urasaki, Ph.D., Postdoctoral Research Associate, Department of Biomedical Sciences, College of Medicine, Roseman University. Project: Identification of novel biomarkers of different forms of fatty liver disease (high-fat diet-induced, genetic mutation-induced, drug-induced) using the cIEF NanoPro 1000 system (see Publications and Patent application).

  • Oscar B. Goodman, Jr., M.D., Ph.D., Associate Professor, Department of Internal Medicine, College of Medicine, Roseman University of Health Sciences, and Clinical Oncologist, Comprehensive Cancer Centers of Nevada (CCCN), Las Vegas & Henderson, NV. Collaboration between Dr. Goodman and Dr. Fiscus first began when Dr. Fiscus was asked to serve as the Mentor on two of Dr. Goodman's grants, a Department of Defense grant and an American Cancer Society grant, while Dr. Fiscus was the Director of Cancer Molecular Biology and Dr. Goodman was a Scientist and Clinician at the Nevada Cancer Institute (NVCI), 2007 - 2010. During this time, the Fiscus Lab was instrumental in helping Dr. Goodman set up his research lab at NVCI, with the Fiscus Lab manager, Mary G. Johlfs, helping to train the early technicians in Dr. Goodman's lab. After leaving NVCI and joining Roseman University of Health Sciences, Dr. Fiscus recruited Dr. Goodman and his two lab members, Dr. Rajana Mita and Dr. Olivia Chao, to join Roseman University to continue their research efforts.