Astrocytic S100B, Blood-Brain Barrier and Neurodegenerative Diseases

Increased life span and expectations of a better quality of life have resulted in a spotlight on neurodegenerative and cardiovascular diseases generally associated with aging. The drive toward evidence-based medicine has necessitated a constant search for objective biomarkers to assay disease onset, progress, and outcomes to make the best clinical decisions. Enhancement of their use depends on the mechanistic understanding of the biomarker’s role in the disease process itself. This chapter focuses on S100B. It is a calcium sensor protein that is primarily astrocytic. While it plays a complex, interlinked role in signaling, serum levels of S100B as a biomarker for clinical decisions is also an area of intense investigation. Both aspects are presented, with an emphasis on the role of S100B in in maintaining a blood-brain barrier, especially in the context of suggesting a unified mechanism for the onset and progression of neurodegenerative diseases

Evolutionary Interrelationships and Insights into Molecular Mechanisms of Functional Divergence: An Analysis of Neuronal Calcium Sensor Proteins

The normal function of any organism, its organizational complexity notwithstanding, depends on the interaction of its proteins with their targets. Thus, analysis of target site interaction is an essential part of all biology. At the protein level, such analyses are critical to both mechanistic knowledge and potential clinical applications such as drug discovery. Approaches to map amino acid residues involved in target site interaction typically are experimental or based on three-dimensional structures obtained through crystallography. Here we test a novel approach that combines phylogenetic analyses with mining of experimental data using neuronal calcium sensor proteins. The proteins fall into three groups based on sequence comparison. One interaction was taken up for analysis from each group. Using the sequence divergence to evaluate the role of amino acids identified experimentally to form the interface with the target, we demonstrate that it is possible to predict residues that are likely to contribute to the specificity of the interaction and, therefore, the functional divergence. Thus, evolutionary analyses of proteins provide an important addition in approaches to generate refined maps of target site interactions in proteins. This approach is especially useful in delineating the functional divergence in a family of closely related proteins

Distinguishing Multiple System Atrophy vs Parkinson’s Disease in an African American Woman

An 87-year-old African American woman with a past medical history of atrial fibrillation on warfarin and peripheral neuropathy with a family history of myasthenia gravis presented to the Emergency Department. The primary reason was loss of consciousness upon standing. The patient was given the diagnosis of hypertension, cervical spinal stenosis, and Parkinson’s Disease. There is little improvement with medications for any of these conditions. Currently, patient has episodes of worsening BP, blackouts, dysphagia, snoring, decreasing voice pitch with trismus. In addition, the patient is positive for dizziness, mild resting tremor in left hand with rigidity, constipation, multiple UTIs and postural instability. The patient and caregiver feel strongly that the care received is less than optimal. The possibility of Multiple System Atrophy, which usually is detected in younger (around age 50) patients, is examined based on differential diagnosis

Pain and Sleep are Associated in Fibromyalgia Patients

This poster explores whether a correlation exists between lack of sleep and fibromyalgia pain

Data on Final Calcium Concentration in Native Gel Reagents Determined Accurately Through Inductively Coupled Plasma Measurements

In this article we present data on the concentration of calcium as determined by Inductively Coupled Plasma (ICP) measurements. Calcium was estimated in the reagents used for native gel electrophoresis of Neuronal Calcium Sensor (NCS) proteins. NCS proteins exhibit calcium-dependent mobility shift in native gels. The sensitivity of this shift to calcium necessitated a precise determination of calcium concentrations in all reagents used. We determined the calcium concentrations in different components used along with the samples in the native gel experiments. These were: 20 mM Tris pH 7.5, loading dye and running buffer, with distilled water as reference. Calcium determinations were through ICP measurements. It was found that the running buffer contained calcium (244 nM) over the blank

Global Comparison of Health Policies Focused on Gestational Diabetes: Recognizing Pertinent Gaps

Introduction: Gestational diabetes mellitus (GDM) is a condition that affects 14.7% of women globally and occurs when a mother develops diabetes during the course of her pregnancy.1 Increased insulin resistance in pregnant mothers can lead to further complications, such as a larger baby, increased risk of the baby developing Type II diabetes, having low blood sugar, and even premature birth, which can lead to respiratory issues. The lack of policies in place to address GDM in various countries worldwide highlights the need for this literature. It is necessary to compile policies that are currently in place and to further discuss how to protect the health of pregnant women who have GDM and their children. Aim: The goals of this project are to compile health policies pertaining to Gestational Diabetes implemented in several countries across the world, to analyze core differences and similarities amongst Gestational Diabetes protocols, and to pinpoint disparities in existing health policies in order to determine areas of policy improvement. Conclusions: There are clear identifiable gaps amongst the different health policies. There are many populations that are predisposed to gestational diabetes due to genetics, past pregnancies, area of residency, socioeconomic status, etc. Future studies should pinpoint areas within a country to be able to develop initiatives that allow reduction in the health gaps seen

Extravasated Brain-Reactive Autoantibodies Perturb Neuronal Surface Protein Expression in Alzheimer\u27s Pathology

Background: Increased blood-brain barrier (BBB) permeability is reported in both the neuropathological and in vivo studies in both Alzheimer’s Disease (AD) and age matched cognitively normal, no cognitive impairment (NCI), subjects. Impaired BBB allows various vascular components such as immunoglobulin G (IgG) to extravasate into the brain and specifically bind to various neuronal surface proteins (NSP), also known as brain reactive autoantibodies (BrABs). This interaction is predicted to further enhance deposition of amyloid plaques. Hypothesis: Interaction between extravasated BrABs and its cognate NSPs lower the expression of that NSPs in AD patients. Methods: We selected Western blotting technique to study the expression of various brain proteins and test our hypothesis. Fresh frozen brain samples of AD and NCI subjects were acquired, and total brain protein was extracted using protocol established in Acharya lab. We also identified various NSPs to study the impact of BrABs-NSPs interactions. Additionally, we investigated the expression of amyloid plaques ((amyloid precursor protein (APP)) and apoptosis (Caspase-3) markers. Specific NSPs examined included the alpha7 nicotinic acetylcholine receptor (α7nAChR) and anti-choline acetyltransferase (ChAT). To image the membranes, fluorescent imaging was used initially, which was later switched to chemiluminescence, after much troubleshooting. Results: Most of the work done through these experiments was focused on establishing a thorough Western blot protocol that can be used to reliably perform these experiments. This involved determining the appropriate primary and secondary antibodies concentrations, loading concentrations, and testing different imaging settings to determine the most ideal image-acquisition conditions. Towards the end of the fellowship, we were successful in developing a protocol to further explore our investigation. Using this protocol, we were able to visualize bands for ChAT, α7nAChR, and caspase – 3. Conclusions: Using this protocol further Western blot experiments can be run to study and compare the expression levels of various NSP in AD and control samples for testing our hypothesi

The Brodmann Area 39/40 of the Brain in Alzheimer’s, Mild Cognitive Impairment, and No Cognitive Impairment Subjects at Advanced Age Demonstrate Comparable Levels of Blood-Brain Barrier Breach

• Alzheimer’s disease (AD) is one of the most common form of dementia • Mild cognitive impairment (MCI), specifically amnestic subtype, more likely to progress to AD • Pathogenesis Theories: o Accumulation of amyloid-beta peptides and neurofibrillary tangles containing hyperphosphorylated neuronal tau protein o Blood Brain Barrier (BBB) dysfunction is associated with AD pathogenesis • Brodmann area 39/40: regions of parietal cortex are responsible for language, spatial cognition, memory retrieval, attention, phonological processing, and emotional processing • Hypothesis: An increased BBB permeability in Brodmann area 39/40 of AD and age-matched MCI and no cognitive impairment (NCI) subject

A Preliminary Report: The Hippocampus and Surrounding Temporal Cortex of Patients With Schizophrenia Have Impaired Blood-Brain Barrier

Schizophrenia (SZ) is one of the most severe forms of mental illness, yet mechanisms remain unclear. A widely established brain finding in SZ is hippocampal atrophy, and a coherent explanation similarly is lacking. Epidemiological evidence suggests increased cerebrovascular and cardiovascular complications in SZ independent of lifestyle and medication, pointing to disease-specific pathology. Endothelial cell contributions to blood-brain barrier (BBB) compromise may influence neurovascular unit and peripheral vascular function, and we hypothesize that downstream functional and structural abnormalities may be explained by impaired BBB

The Changing Landscape for Stroke\ua0Prevention in AF: Findings From the GLORIA-AF Registry Phase 2

Background GLORIA-AF (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients with Atrial Fibrillation) is a prospective, global registry program describing antithrombotic treatment patterns in patients with newly diagnosed nonvalvular atrial fibrillation at risk of stroke. Phase 2 began when dabigatran, the first non\u2013vitamin K antagonist oral anticoagulant (NOAC), became available. Objectives This study sought to describe phase 2 baseline data and compare these with the pre-NOAC era collected during phase 1. Methods During phase 2, 15,641 consenting patients were enrolled (November 2011 to December 2014); 15,092 were eligible. This pre-specified cross-sectional analysis describes eligible patients\u2019 baseline characteristics. Atrial fibrillation disease characteristics, medical outcomes, and concomitant diseases and medications were collected. Data were analyzed using descriptive statistics. Results Of the total patients, 45.5% were female; median age was 71 (interquartile range: 64, 78) years. Patients were from Europe (47.1%), North America (22.5%), Asia (20.3%), Latin America (6.0%), and the Middle East/Africa (4.0%). Most had high stroke risk (CHA2DS2-VASc [Congestive heart failure, Hypertension, Age  6575 years, Diabetes mellitus, previous Stroke, Vascular disease, Age 65 to 74 years, Sex category] score  652; 86.1%); 13.9% had moderate risk (CHA2DS2-VASc = 1). Overall, 79.9% received oral anticoagulants, of whom 47.6% received NOAC and 32.3% vitamin K antagonists (VKA); 12.1% received antiplatelet agents; 7.8% received no antithrombotic treatment. For comparison, the proportion of phase 1 patients (of N = 1,063 all eligible) prescribed VKA was 32.8%, acetylsalicylic acid 41.7%, and no therapy 20.2%. In Europe in phase 2, treatment with NOAC was more common than VKA (52.3% and 37.8%, respectively); 6.0% of patients received antiplatelet treatment; and 3.8% received no antithrombotic treatment. In North America, 52.1%, 26.2%, and 14.0% of patients received NOAC, VKA, and antiplatelet drugs, respectively; 7.5% received no antithrombotic treatment. NOAC use was less common in Asia (27.7%), where 27.5% of patients received VKA, 25.0% antiplatelet drugs, and 19.8% no antithrombotic treatment. Conclusions The baseline data from GLORIA-AF phase 2 demonstrate that in newly diagnosed nonvalvular atrial fibrillation patients, NOAC have been highly adopted into practice, becoming more frequently prescribed than VKA in Europe and North America. Worldwide, however, a large proportion of patients remain undertreated, particularly in Asia and North America. (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients With Atrial Fibrillation [GLORIA-AF]; NCT01468701