Digital Commons Network^™

Genetic Determinants Of Amidating Enzyme Activity And Its Relationship With Metal Cofactors In Human Serum, Eric D. Gaier, Alison Kleppinger, Martina Ralle, Jonathan Covault, Richard E. Mains, Anne M. Kenny, Betty A. Eipper

UCHC Articles - Research

Abstract

BACKGROUND:

α-amidation is a final, essential step in the biosynthesis of about half of all peptide hormones and neurotransmitters. Peptidylglycine α-amidating monooxygenase (PAM), with enzymatic domains that utilize Cu and Zn, is the only enzyme that catalyzes this reaction. PAM activity is detected in serum, but its significance and utility as a clinical biomarker remain unexplored.

METHODS:

We used well-established enzymatic assays specific for the peptidylglycine-α -hydroxylating monooxygenase (PHM) and peptidyl-α-hydroxyglycine α-amidating lyase (PAL) domains of PAM to quantify amidating activity in the sera of 144 elderly men. Relationships between PHM and PAL activity and serum levels of their …

Simple Signaling Molecules For Inductive Bone Regenerative Engineering, Bret D. Ulery, Stephen J. Nelson, Meng Deng, Kevin W. H. Lo, Yusuf M. Khan, Cato T. Laurencin

UCHC Articles - Research

Abstract

With greater than 500,000 orthopaedic procedures performed in the United States each year requiring a bone graft, the development of novel graft materials is necessary. We report that some porous polymer/ceramic composite scaffolds possess intrinsic osteoinductivity as shown through their capacity to induce in vivo host osteoid mineralization and in vitro stem cell osteogenesis making them attractive synthetic bone graft substitutes. It was discovered that certain low crystallinity ceramics partially dissociate into simple signaling molecules (i.e., calcium and phosphate ions) that induce stem cells to endogenously produce their own osteoinductive proteins. Review of the literature has uncovered a variety …

Human Esc-Derived Mscs Outperform Bone Marrow Mscs In The Treatment Of An Eae Model Of Multiple Sclerosis, Xiaofang Wang, Kumiko Ijichi, Debayon Paul, Adam S. Lazorchak, Joel S. Pachter, Stephen J. Crocker, Ren-He Xu

UCHC Articles - Research

Current therapies for multiple sclerosis (MS) are largely palliative, not curative. Mesenchymal stem cells (MSCs) harbor regenerative and immunosuppressive functions, indicating a potential therapy for MS, yet the variability and low potency of MSCs from adult sources hinder their therapeutic potential. MSCs derived from human embryonic stem cells (hES-MSCs) may be better suited for clinical treatment of MS because of their unlimited and stable supply. Here, we show that hES-MSCs significantly reduce clinical symptoms and prevent neuronal demyelination in a mouse experimental autoimmune encephalitis (EAE) model of MS, and that the EAE disease-modifying effect of hES-MSCs is significantly greater than …

Tooth Loss Strongly Associates With Malnutrition In Chronic Kidney Disease, Efthimia Effie Ioannidou, Helen Swede

UCHC Articles - Research

Background

In chronic kidney disease (CKD), inadequate nutritional intake, inflammation, and increased oxidative stress have been the major contributing factors in malnutrition pathogenesis. However, there is still a paucity of evidence assessing the magnitude of the effect of tooth loss on malnutrition in CKD populations. The authors hypothesize that among patients with CKD, tooth loss may affect nutritional status, using the National Health and Nutrition Examination Survey 1988 to 1994 (NHANES III).

Methods

Glomerular filtration rate (GFR) was estimated based on cystatin C levels using the relevant equation. Urinary albumin-to-creatinine ratio (albuminuria) was calculated in milligrams per gram with a …

Prize Contingency Management For Smoking Cessation: A Randomized Trial, Nancy M. Petry, Sheila M. Alessi

UCHC Articles - Research

Abstract

Background

Adjunctive behavioral smoking cessation treatments have the potential to improve outcomes beyond standard care. The present study had two aims: 1) compare standard care (SC) for smoking (four weeks of brief counseling and monitoring) to SC plus prize-based contingency management (CM), involving the chance to earn prizes on days with demonstrated smoking abstinence (carbon monoxide (CO) ≤6ppm); and 2) compare the relative efficacy of two prize reinforcement schedules - one a traditional CM schedule, and the second an early enhanced CM schedule providing greater reinforcement magnitude in the initial week of treatment but equal overall reinforcement.

Methods

Participants …

Increased Mitochondrial Biogenesis Preserves Intestinal Stem Cell Homeostasis And Contributes To Longevity In Indy Mutant Flies, Ryan P. Rogers, Blanka Rogina

UCHC Articles - Research

The Drosophila Indy (I'm Not Dead Yet) gene encodes a plasma membrane transporter of Krebs cycle intermediates, with robust expression in tissues associated with metabolism. Reduced INDY alters metabolism and extends longevity in a manner similar to caloric restriction (CR); however, little is known about the tissue specific physiological effects of INDY reduction. Here we focused on the effects of INDY reduction in the Drosophila midgut due to the importance of intestinal tissue homeostasis in healthy aging and longevity. The expression of Indy mRNA in the midgut changes in response to aging and nutrition. Genetic reduction of Indy expression increases …

An Investigation Of The Association Between Omega 3 Fa And Bone Mineral Density Among Older Adults: Results From The National Health And Nutrition Examination Survey Years 2005– 2008, Kelsey M. Mangano, Jane E. Kerstetter, Anne M. Kenny, Stephen J. Walsh

UCHC Articles - Research

Summary

The relation of omega 3 fatty acids (n-3 FA) with bone mineral density (BMD) was assessed among adults >60 years; NHANES data (2005–2008). The association of dietary n-3 FA with measures of hip BMD was equivocal, but n-3 FA supplement use was significantly associated with higher spine BMD—a finding that deserves further study.

Introduction

Associations between polyunsaturated fatty acids and bone mineral density are not well understood.

Purpose

To evaluate the cross-sectional relation between dietary omega 3 fatty acid intake (specifically docosahexaenoic acid, eicosapentaenoic acid, and octadecatetraenoic) and BMD at the hip and spine among older adults.

Methods

Omega …

Resolution Of Central Nervous System Astrocytic And Endothelial Sources Of Ccl2 Gene Expression During Evolving Neuroinflammation, Bandana Shrestha, Shujun Ge, Joel S. Pachter

UCHC Articles - Research

Background

The chemokine CCL2 is a critical mediator of neuroinflammation in diseases such as multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). CCL2 drives mononuclear cell infiltration into the central nervous system (CNS), alters expression and distribution of microvascular endothelial tight junction proteins, and disrupts the blood–brain and blood-spinal cord barriers. Immunohistochemistry has consistently revealed astrocytes to be a source of this chemokine during neuroinflammation, while providing less uniform evidence that CNS endothelial cells may also express CCL2. Moreover, the relative contributions of these cell types to the CNS pool of CCL2 during MS/EAE are unclear and …

Impact Of Macrophage Deficiency And Depletion On Continuous Glucose Monitoring In Vivo, Ulrike Klueh, Yi Qiao, Jackman T. Frailey, Donald L. Kreutzer

UCHC Articles - Research

Although it is assumed that macrophages (MQ) have a major negative impact on continuous glucose monitoring (CGM), surprisingly there is no data in the literature to directly support or refute the role of MQ or related foreign body giant cells in the bio-fouling of glucose sensors in vivo. As such, we developed the hypothesis that MQ are key in controlling glucose sensor performance and CGM in vivo and MQ deficiencies or depletion would enhance CGM. To test this hypothesis we determined the presence/distribution of MQ at the sensor tissue interface over a 28-day time period using F4/80 antibody and immunohistochemical …

Adequate Dietary Protein Is Associated With Better Physical Performance Among Post-Menopausal Women 60–90 Years, Leah Gregorio, Jennifer L. Brindisi, Alison Kleppinger, K. M. Mangano, J. D. Bihuniak, Anne M. Kenny, J. E. Kerstetter

UCHC Articles - Research

Objectives

Sarcopenia, the involuntary loss of skeletal muscle with age, affects up to one-quarter of older adults. Evidence indicates a positive association between dietary protein intake and lean muscle mass and strength among older persons, but information on dietary protein’s effect on physical performance in older adults has received less attention.

Design

Cross-sectional observational analysis of the relationship of dietary protein on body composition and physical performance.

Setting

Clinical research center.

Participants

387 healthy women aged 60 – 90 years (mean 72.7 ± 7.0 y).

Measurements

Measures included body composition (fat-free mass, appendicular skeletal mass and fat mass) via dual …

Cell-Selective Knockout And 3d Confocal Image Analysis Reveals Separate Roles For Astrocyte- And Endothelial-Derived Ccl2 In Neuroinflammation, Debayon Paul, Shujun Ge, Yen Lemire, Evan R. Jellison, David R. Serwanski, Joel S. Pachter

UCHC Articles - Research

Background

Expression of chemokine CCL2 in the normal central nervous system (CNS) is nearly undetectable, but is significantly upregulated and drives neuroinflammation during experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis which is considered a contributing factor in the human disease. As astrocytes and brain microvascular endothelial cells (BMEC) forming the blood–brain barrier (BBB) are sources of CCL2 in EAE and other neuroinflammatory conditions, it is unclear if one or both CCL2 pools are critical to disease and by what mechanism(s).

Methods

Mice with selective CCL2 gene knockout (KO) in astrocytes (Astro KO) or …

Complex Mental Health Sequelae Of Psychological Trauma Among Women In Prenatal Care, Julian D. Ford

UCHC Articles - Research

Pregnancy is a critical time to identify and address maternal mental health problems, for the health of both mother and child. Pregnant women with histories of exposure to interpersonal psychological trauma may experience a range of mental health problems including but not limited to posttraumatic stress disorder (PTSD). In a community sample of 1,581 pregnant women, 25% reported symptoms consistent with at least one of six syndromes, including PTSD, major depressive disorder (MDD), generalized anxiety disorder (GAD), or clinically significant dissociation, somatization, or affect dysregulation. Six sub-groups with distinct mental health problem profiles were identified by cluster analysis. Controlling for …

A Systems Biology Approach To Iron Metabolism, Reinhard C. Laubenbacher, Suzy V. Torti

UCHC Articles - Research

Iron is critical to the survival of almost all living organisms. However, inappropriately low or high levels of iron are detrimental and contribute to a wide range of diseases. Recent advances in the study of iron metabolism have revealed multiple intricate pathways that are essential to the maintenance of iron homeostasis. Further, iron regulation involves processes at several scales, ranging from the subcellular to the organismal. This complexity makes a systems biology approach crucial, with its enabling technology of computational models based on a mathematical description of regulatory systems. Systems biology may represent a new strategy for understanding imbalances in …

Expression Of Phosphodiesterase 6 (Pde6) In Human Breast Cancer Cells, Hongli Dong, Kevin P. Claffey, Stefan Brocke, Paul M. Epstein

UCHC Articles - Research

Considerable epidemiological evidence demonstrates a positive association between artificial light at night (LAN) levels and incidence rates of breast cancer, suggesting that exposure to LAN is a risk factor for breast cancer. There is a 30-50% higher risk of breast cancer in the highest LAN exposed countries compared to the lowest LAN countries, and studies showing higher incidence of breast cancer among shift workers exposed to more LAN have led the International Agency for Research on Cancer to classify shift work as a probable human carcinogen. Nevertheless, the means by which light can affect breast cancer is still unknown. In …

One-Day Treatment Of Small Molecule 8-Bromo-Cyclic Amp Analogue Induces Cell-Based Vegf Production For In Vitro Angiogenesis And Osteoblastic Differentiation, Kevin W.-H. Lo, Ho Man Kan, Keith A. Gagnon, Cato T. Laurencin

UCHC Articles - Research

Small molecule based regenerative engineering is emerging as a promising strategy for regenerating bone tissue. Small molecule cAMP analogues have been proposed as novel biofactors for bone repair and regeneration, and while promising, the effect that these small molecules have on angiogenesis, a critical requirement for successful bone regeneration, is still unclear. Our previous research demonstrated that the small molecule cAMP analogue 8-bromoadenosine-3’,5’-cyclic monophosphate (8-Br-cAMP) was able to promote initial osteoblast adhesion on a polymeric scaffold via cAMP signaling cascades. Here, we report that 8-Br-cAMP is capable of inducing in vitro cell-based VEGF production for angiogenesis promotion. We first demonstrated …

Microglial Responses After Ischemic Stroke And Intracerebral Hemorrhage, Roslyn A. Taylor, Lauren H. Sansing

UCHC Articles - Research

Stroke is a leading cause of death worldwide. Ischemic stroke is caused by blockage of blood vessels in the brain leading to tissue death, while intracerebral hemorrhage (ICH) occurs when a blood vessel ruptures, exposing the brain to blood components. Both are associated with glial toxicity and neuroinflammation. Microglia, as the resident immune cells of the central nervous system (CNS), continually sample the environment for signs of injury and infection. Under homeostatic conditions, they have a ramified morphology and phagocytose debris. After stroke, microglia become activated, obtain an amoeboid morphology, and release inflammatory cytokines (the M1 phenotype). However, microglia can …

Preosteocytes/Osteocytes Have The Potential To Dedifferentiate Becoming A Source Of Osteoblasts, Elena Torreggiani, Brya G. Matthews, Slavica Pejda, Igor Matic, Danka Grcevic, Mark C. Horowitz, Ivo Kalajzic

UCHC Articles - Research

Presently there is no clear evidence for the ability of mature osteogenic lineage cells to dedifferentiate. In order to identify and trace mature osteogenic lineage cells, we have utilized transgenic mouse models in which the dentin matrix protein 1 (Dmp1) promoter drives expression of GFP (active marker) or Cre recombinase (historic label) in preosteocytes/osteocytes. In long bone chip outgrowth cultures, in which cells on the bone surface were enzymatically removed, cells with previous activity of the Dmp1 promoter migrated onto plastic and down-regulated Dmp1-GFP expression. Dmp1Cre-labeled cells from these cultures had the potential to re-differentiate into the osteogenic lineage, while …

Can Stem Cells Be Used To Generate New Lungs? Ex Vivo Lung Bioengineering With Decellularized Whole Lung Scaffolds, Todd J. Jensen, Eric D. Girard, Christine M. Finck

UCHC Articles - Research

For patients with end-stage lung diseases, lung transplantation is the only available therapeutic option. However, the number of suitable donor lungs is insufficient and lung transplants are complicated by significant graft failure and complications of immunosuppressive regimens. An alternative to classic organ replacement is desperately needed. Engineering of bioartificial organs using either natural or synthetic scaffolds is an exciting new potential option for generation of functional pulmonary tissue for human clinical application. Natural organ scaffolds can be generated by decellularization of native tissues; these acellular scaffolds retain the native organ ultrastructure and can be seeded with autologous cells toward the …

Osterix-Cre Labeled Progenitor Cells Contribute To The Formation And Maintenance Of The Bone Marrow Stroma, Yaling Liu, Sara Strecker, Liping Wang, Mark S. Kronenberg, Wen Wang, David W. Rowe, Peter F. Maye

UCHC Articles - Research

We have carried out fate mapping studies using Osterix-EGFPCre and Osterix-CreERt animal models and found Cre reporter expression in many different cell types that make up the bone marrow stroma. Constitutive fate mapping resulted in the labeling of different cellular components located throughout the bone marrow, whereas temporal fate mapping at E14.5 resulted in the labeling of cells within a region of the bone marrow. The identity of cell types marked by constitutive and temporal fate mapping included osteoblasts, adipocytes, vascular smooth muscle, perineural, and stromal cells. Prolonged tracing of embryonic precursors labeled at E14.5dpc revealed the continued …

Utilization Of Transgenic Models In Evaluation Of Osteogenic Differentiation Of Embryonic Stem Cells, Dario Repic, Elena Torreggiani, Tiziana Franceschetti, Brya G. Matthews, Alexander C. Lichtler, Ivo Kalajzic

UCHC Articles - Research

Previous studies reported that embryonic stem cells (ESCs) can be induced to differentiate into cells showing a mature osteoblastic phenotype by culturing them under osteo-inductive conditions. It is probable that osteogenic differentiation requires that ESCs undergo differentiation through an intermediary step involving a mesenchymal lineage precursor. Based on our previous studies indicating that adult mesenchymal progenitor cells express αSMA, we have generated ESCs from transgenic mice in which an αSMA promoter directs the expression of red fluorescent protein (RFP) to mesenchymal progenitor cells. To track the transition of ESC-derived MSCs into mature osteoblasts, we have utilized a bone-specific fragment of …

The Effects Of Estrogen In Ischemic Stroke, Louise D. Mccullough, Edward C. Koellhoffer

UCHC Articles - Research

Stroke is a leading cause of death and the most common cause of long-term disability in the USA. Women have a lower incidence of stroke compared with men throughout most of the lifespan which has been ascribed to protective effects of gonadal steroids, most notably estrogen. Due to the lower stroke incidence observed in pre-menopausal women and robust preclinical evidence of neuroprotective and anti-inflammatory properties of estrogen, researchers have focused on the potential benefits of hormones to reduce ischemic brain injury. However, as women age, they are disproportionately affected by stroke, coincident with the loss of estrogen with menopause. The …

Knockout Of Vascular Early Response Gene Worsens Chronic Stroke Outcomes In Neonatal Mice, Mehwish A. Mirza, Lori A. Capozzi, Yan Xu, Louise D. Mccullough, Fudong Liu

UCHC Articles - Research

Vascular early response gene (Verge) is a novel immediate early gene that is highly expressed during developmental angiogenesis and after ischemic insults in adult brain. However, the role of Verge after neonatal injury is not known. In the present study, we investigated the hypothesis that Verge contributes to vascular remodeling and tissue repair after neonatal ischemic injury. The Rice–Vanucci model (RVM) was employed to induce neonatal stroke in both Verge knockout (KO) and wild-type (WT) postnatal day 10 (P10) mice. Histological and behavioral outcomes at acute (24 h), subacute (7 days) and chronic (30 days) phases were evaluated. Angiogenesis, neurogenesis, …

Transgenic Sickle Cell Disease Mice Have High Mortality And Dysregulated Immune Responses After Vaccination, Steven M. Szczepanek, Eric R. Secor Jr, Sonali J. Bracken, Linda Guernsey, Ektor Rafti, Adam Matson, Roger S. Thrall

UCHC Articles - Research

Background

Children with sickle cell disease (SCD) are susceptible to recurrent infections, which are often life threatening and necessitate frequent vaccinations. Given the altered baseline immunity and proinflammatory state associated with SCD, we sought to determine the relative safety and efficacy of vaccination in transgenic SCD mice.

Methods

Eight week-old SCD mice were vaccinated with ovalbumin (OVA) and aluminum hydroxide weekly for three weeks by the intraperitoneal (IP) or intramuscular (IM) route. One week after the third vaccination, serum cytokines/chemokines, immunoglobulins, and bronchoalveolar lavage (BAL) fluid cytokines were measured.

Results

Only SCD mice were prone to mortality associated with vaccination …

Ptpn11 Deletion In A Novel Cartilage Cell Causes Metachondromatosis By Activating Hedgehog Signaling, Qian Wu

UCHC Articles - Research

SHP2, encoded by PTPN11, is required for survival, proliferation and differentiation of various cell types^1,2. Germ line activating mutations in PTPN11 cause Noonan Syndrome, while somatic PTPN11 mutations cause childhood myeloproliferative disease and contribute to some solid tumors. Recently, heterozygous inactivating mutations in PTPN11 were found in metachondromatosis, a rare inherited disorder featuring multiple exostoses, endochondromas, joint destruction and bony deformities^3,4. The detailed pathogenesis of this disorder has remained unclear. Here, we used a conditional knockout allele (Ptpn11^fl) and Cre recombinase (Cre) transgenic mice to delete Ptpn11 specifically in …

The Transcriptomics To Proteomics Of Hair Cell Regeneration: Looking For A Hair Cell In A Haystack, Gopinath Rajadinakaran

UCHC Articles - Research

Mature mammals exhibit very limited capacity for regeneration of auditory hair cells, while all non-mammalian vertebrates examined can regenerate them. In an effort to find therapeutic targets for deafness and balance disorders, scientists have examined gene expression patterns in auditory tissues under different developmental and experimental conditions. Microarray technology has allowed the large-scale study of gene expression profiles (transcriptomics) at whole-genome levels, but since mRNA expression does not necessarily correlate with protein expression, other methods, such as microRNA analysis and proteomics, are needed to better understand the process of hair cell regeneration. These technologies and some of the results of …

Subtelomeric Hotspots Of Aberrant 5-Hydroxymethylcytosine-Mediated Epigenetic Modifications During Reprogramming To Pluripotency, Stormy J. Chamberlain, I-Ping Chen

UCHC Articles - Research

Mammalian somatic cells can be directly reprogrammed into induced pluripotent stem cells (iPSCs) by introducing defined sets of transcription factors. Somatic cell reprogramming involves epigenomic reconfiguration, conferring iPSCs with characteristics similar to embryonic stem cells (ESCs). Human ES cells contain 5-hydroxymethylcytosine (5hmC), which is generated through the oxidation of 5-methylcytosine by the TET enzyme family. Here we show that 5hmC levels increase significantly during reprogramming to human iPSCs mainly due to TET1 activation, and this hydroxymethylation change is critical for optimal epigenetic reprogramming, but does not compromise primed pluripotency. Compared with hES cells, we find iPS cells tend to form …

Mmp-3 Mediates Psychosine-Induced Globoid Cell Formation: Implications For Leukodystrophy Pathology, Kumiko Ijichi, Graham D. Brown, Craig S. Moore, Paige N. Winokur, Roberto Pagarigan, Stephen J. Crocker

UCHC Articles - Research

Globoid cell leukodystrophy (GLD) or Krabbe disease, is a fatal demyelinating disease attributed to mutations in the galactocerebrosidase (GALC) gene. Loss of function mutations in GALC result in accumulation of the glycolipid intermediate, galactosylsphingosine (psychosine). Due to the cytotoxicity of psychosine, it has been hypothesized that accumulated psychosine underlie the pathophysiology of GLD. However, the cellular mechanisms of GLD pathophysiology remain unclear. Globoid cells, multinucleated microglia/macrophages in the central nervous system (CNS), are a defining characteristic of GLD. Here we report that exposure of primary glial cultures to psychosine induces the expression and the production of matrix metalloproteinase …

Racial And Ethnic Disparities In Diabetes Complications In The Northeastern United States: The Role Of Socioeconomic Status, Julie A. Wagner

UCHC Articles - Research

The role of socioeconomic status (SES) in explaining racial/ethnic disparities in diabetes remains unclear. We investigated disparities in self-reported diabetes complications, and the role of macro (e.g., income, education) and micro (e.g., ‘owning a home’ or ‘having a checking account’) SES indicators in explaining these differences. The sample included individuals with a diagnosis of diabetes (N=795) who were on average 55 years old, and 55.6% non-Hispanic White, 25.0% African American, and 19.4% Hispanic. Approximately 8% reported nephropathy, 35% reported retinopathy, and 16% reported cardiovascular disease. There were significant disparities in the rates of complications among non-Hispanic White, African American and …

The Sh2 Domain Interaction Landscape, Kazuya Machida, Christopher M. Thompson, Bruce J. Mayer

UCHC Articles - Research

Members of the SH2 domain family modulate signal transduction by binding to short peptides containing phosphorylated tyrosines. Each domain displays a distinct preference for the sequence context of the phosphorylated residue. We have developed a new high-density peptide chip technology that allows probing the affinity of most SH2 domains for a large fraction of the entire complement of tyrosine phosphopeptides in the human proteome. Using this technique we have experimentally identified thousands of putative SH2- peptide interactions for more than 70 different SH2 domains. By integrating this rich data set with orthogonal context-specific information, we have assembled an SH2 mediated …

Computationally Designed Peptide Inhibitors Against The Ubiquitin E3 Ligase Scf Fbx4, Bing Hao

UCHC Articles - Research

Telomerase is a reverse transcriptase that maintains telomere length. Telomerase activity is suppressed in somatic cells such that telomere attrition triggers replicative senescence or apop-tosis. In cancer cells, telomerase is up-regulated or reactivated, effectively making the cell immortal. Previous studies have shown that telomerase activity positively correlates with unfavorable cancer prognosis. Since it was discovered that activation of telomerase is a rate-limiting step in carcinogenesis, telomerase has gained much interest as a drug target. Both screening and structure-based methods have been extensively employed to identify small molecule leads that can selectively disrupt telomerase activity. Strategies commonly used to target telomerase …