Medicine and Health Sciences Commons^™

Optimizing Production Of Fc-Amidated Peptides By Chinese Hamster Ovary Cells, Kristina Carlson, Richard E. Mains, Betty A. Eipper

UCHC Articles - Research

Background

Amidation of the carboxyl terminal of many peptides is essential for full biological potency, often increasing receptor binding and stability. The single enzyme responsible for this reaction is peptidylglycine α-amidating monooxygenase (PAM: EC 1.14.17.3), a copper- and ascorbate-dependent Type I membrane protein.

Methods

To make large amounts of high molecular weight amidated product, Chinese hamster ovary (CHO) cells were engineered to express exogenous PAM. To vary access of the enzyme to its substrate, exogenous PAM was targeted to the endoplasmic reticulum, trans-Golgi network, endosomes and lysosomes or to the lumen of the secretory pathway.

Results

PAM was equally …

Pertinacity Issue 1 Volume 2, Yuri Hosokawa

Pertinacity

No abstract provided.

Phosphorylation State-Dependent Interaction Between Akap7Δ/Γ And Phospholamban Increases Phospholamban Phosphorylation., Marc Rigatti, Andrew V. Le, Claire Gerber, Ion I. Moraru, Kimberly L. Dodge-Kafka

UCHC Articles - Research

Changes in heart rate and contractility in response to sympathetic stimulation occur via activation of cAMP dependent protein kinase A (PKA), leading to phosphorylation of numerous substrates that alter Ca²⁺ cycling. Phosphorylation of these substrates is coordinated by A-kinase anchoring proteins (AKAPs), which recruit PKA to specific substrates [1]. Phosphorylation of the PKA substrate phospholamban (PLB) is a critical determinant of Ca²⁺ re-entry into the sarcoplasmic reticulum and is coordinated by AKAP7δ/γ [2,3]. Here, we further these findings by showing that phosphorylation of PLB requires interaction with AKAP7δ/γ and that this interaction …

Cloning And Variation Of Ground State Intestinal Stem Cells, Lane H. Wilson, Francisco A. Sylvester, Jeffrey S. Hyams, Thomas Devers, Wa Xian

UCHC Articles - Research

Stem cells of the gastrointestinal tract, pancreas, liver, and other columnar epithelia collectively resist cloning in their elemental states. Here we demonstrate the cloning and propagation of highly clonogenic, “ground state” stem cells of the human intestine and colon. We show that derived stem cell pedigrees sustain limited copy number and sequence variation despite extensive serial passaging and display exquisitely precise, cell-autonomous commitment to epithelial differentiation consistent with their origins along the intestinal tract. This developmentally patterned and epigenetically maintained commitment of stem cells likely enforces the functional specificity of the adult intestinal tract. Using clonally-derived colonic epithelia, we show …

Association Of The Lipoprotein Receptor Scarb1 Common Missense Variant Rs4238001 With Incident Coronary Heart Disease, Annabelle Rodriguez-Oquendo

UCHC Articles - Research

Background

Previous studies in mice and humans have implicated the lipoprotein receptor SCARB1 in association with atherosclerosis and lipid levels. In the current study, we sought to examine association of SCARB1 missense single nucleotide polymorphism (SNP) rs4238001 with incident coronary heart disease (CHD).

Methods and Results

Genotypes for rs4238001 were imputed for 2,319 White, 1,570 African American, and 1,292 Hispanic-American MESA participants using the 1,000 Genomes reference set. Cox proportional hazards models were used to determine association of rs4238001 with incident CHD, with adjustments for age, sex, study site, principal components of ancestry, body mass index, diabetes status, serum creatinine, …

Pertinacity Volume 1 Issue 1, Yuri Hosokawa

Pertinacity

No abstract provided.

Manipulation Of The Microbiome And Its Impact On Functional Recovery Following Ischemic Stroke, Michal Jandzinski

Honors Scholar Theses

Each year, nearly 800,000 individuals residing in the United States will have a stroke. Of these, about 130,000 cases will prove fatal while many of the survivors will be forced to live with disability for the remainder of their lives. Out of all strokes over 87% are ischemic strokes. The widespread incidence of this debilitating condition costs the United States an estimated $36.5 billion dollars every single year. Despite this, clinicians are armed with very little to combat the disease. Recent research developments have brought about the rise in awareness about the importance of the microbiome, the various gut flora …

Pathological Effects Of Repeated Concussive Tbi In Mouse Models: Periventricular Damage And Ventriculomegaly, Richard H. Wolferz Jr.

Honors Scholar Theses

Repeated concussive traumatic brain injury (rcTBI) is the most prominent form of head injury affecting the brain, with an estimated 1.7 million Americans affected each year (Kuhn 2012). Neurologists have been concerned about the danger of repeated head impacts since the 1920’s, but researchers have only begun to understand the long-term effects of rcTBI (McKee 2009). Although symptoms can be as mild as dizziness, current research suggests that multiple concussions can lead to a progressive degenerative brain disease known as chronic traumatic encephalopathy (CTE) (Luo 2008, McKee 2009, Kane 2013). Research on the brain is just beginning to scratch the …

The Development Of Skeletal Muscle In Young Horses: An Ultrasonography And Satellite Cell Approach, Emma K. Lavigne

Honors Scholar Theses

Muscle growth in young horses is characterized by an increase in muscle cross-sectional area, which can be accomplished through the activation and differentiation of satellite cells. Satellite cells can be stimulated or inhibited in response to different cytokines and growth factors and are key mediators of muscle hypertrophy and regeneration. The aim of this study was to investigate the growth of the longissimus dorsi (LD) muscle in horses under 5 years of age and to obtain preliminary data on satellite cell behavior in foals. The area, width, height, and subcutaneous fat were measured using ultrasonography at 6-month increments over the …

Oscillatory Activity In The Subthalamic Nucleus And Motor Cortex In A Pharmacological Model Of Parkinsonian Tremor, Aileen F. Haque

Honors Scholar Theses

Parkinson’s Disease (PD) is a motor disorder with symptoms including resting tremor, akinesia, bradykinesia, and rigidity. A major neuropathological feature of PD is degeneration of nigrostriatal dopamine (DA) neurons. The resulting DA depletions lead to the production of severe motor deficits. Pharmacological agents that reduce DA transmission can also induce these motor abnormalities. In addition to the involvement of DA, drugs acting on acetylcholine, namely cholinomimetics, can induce or exacerbate Parkinsonian symptoms. In humans, one of the main motor symptoms associated with PD is resting tremor, occurring at a frequency of 3-7 Hz. This can be modeled in rodents using …

Proteo-Lipobeads For The Oriented Encapsulation Of Membrane Proteins, Leslie M. Loew

UCHC Articles - Research

As a surrogate of the life cell, proteo-lipobeads are presented, encapsulating functional membrane proteins in a strict orientation into a lipid bilayer. Assays can be performed just as on life cells, for example using fluorescence measurements. As a proof of concept, we have demonstrated proton transport through cytochrome c oxidase.

Vasculogenesis And Angiogenesis In Vegf Receptor-1 Deficient Mice, Vivienne C. Ho, Guo-Hua Fong

UCHC Articles - Research

Vascular endothelial growth factor receptor-1 (VEGFR-1)/Flt-1 is a transmembrane tyrosine kinase receptor for VEGF-A, VEGF-B, and placental growth factor (PlGF). VEGFR-1 is an enigmatic molecule whose precise role in postnatal angiogenesis remains controversial. Although many postnatal and adult studies have been performed by manipulating VEGFR-1 ligands, including competitive binding by truncated VEGFR-1 protein, neutralization by antibodies, or specific ligand overexpression or knockout, much less is known at the level of the receptor per se, especially in vivo. Perplexingly, while VEGFR-1 negatively regulates endothelial cell differentiation during development, it has been implied in promoting angiogenesis under certain conditions in adult tissues, …

Imaging Submillisecond Membrane Potential Changes From Individual Regions Of Single Axons, Dendrites And Spines, Srdjan D. Antic

UCHC Articles - Research

A central question in neuronal network analysis is how the interaction between individual neurons produces behavior and behavioral modifications. This task depends critically on how exactly signals are integrated by individual nerve cells functioning as complex operational units. Regional electrical properties of branching neuronal processes which determine the input-output function of any neuron are extraordinarily complex, dynamic, and, in the general case, impossible to predict in the absence of detailed measurements. To obtain such a measurement one would, ideally, like to be able to monitor, at multiple sites, subthreshold events as they travel from the sites of origin (synaptic contacts …