Determination of Nuclear Factor-Kappa B Activation in Cultured Renal Epithelial Cells and Cardiac Myocytes Exposed to Cocaine and Morphine

Nuclear factor-kappa B (NF-kB) is an important transcription factor that participates in the genetic regulation of inflammation in many tissues. Cocaine and heroin are drugs of abuse associated with renal and cardiac pathology. The purpose of this study was to test the hypothesis that exposure to cocaine or morphine (a metabolite of heroin) would activate NF-kB in renal epithelial cells (COS-7) and cardiac myocytes (H9c2) grown in culture. COS-7 and H9c2 cells were co-transfected with an experimental reporter specific for NF-kB activation and a control reporter with constitutive activity. A dual-luciferase assay was used to determine levels of NF-kB activation after exposure to increasing concentrations of cocaine or morphine. It was determined that 10-2 M cocaine activated NF-kB in both cell types with or without I-kappa B (IkB) overexpression. Cocaine at 10-4 M also activated NF-kB in H9c2 cells, but not in COS-7 cells. Morphine was unable to activate NF-kB in either cell type at any of the concentrations tested. We conclude that cocaine, but not morphine, activates NF-kB in both COS-7 and H9c2 cells grown in culture

The Effects of Cocaine and Ecstasy on Cardiac Myocytes and the Intact Myocardium

Cocaine and ecstasy are widely used illicit drugs. Both drugs have undergone intense scrutiny as information regarding their side-effects has become available. One important yet incomplete area of investigation pertains to their effects on the heart. The purpose of the current studies was to test the hypothesis that exposure to cocaine or ecstasy will adversely affect cellular homeostasis and normal heart function. Cultured cardiac myocytes (H9c2) and New Zealand White rabbits (Oryctolagus cuniculus) were used to measure the responses to various concentrations of cocaine or ecstasy at both the cellular and intact organ system levels. We observed that cocaine and ecstasy significantly altered several homeostatic parameters including reactive oxygen species generation, intracellular calcium balance, NF-κB activity, gene expression, and left ventricular function. We conclude that cocaine and ecstasy are detrimental to the myocardium of the heart, causing several disturbances with pathological potential

Selling Aloha: The Fight for Legal Protections Over Native Hawaiian Culture

In 2018, a Chicago-based restaurant attempted to enforce a registered trademark of “Aloha Poke” by sending cease-and-desist letters to small businesses with names containing some variation of the phrase. Most of those businesses were owned by Native Hawaiians, causing an uproar due to the terms “aloha” and “poke” having strong ties to traditional Hawaiian culture. Known as the Aloha Poke case, it brought attention to the fact that the United States currently has no definite legal framework to protect the cultural heritage of Native Hawaiians, much less their intangible cultural heritage. This Note addresses the lack of federal recognition granted to Native Hawaiians and how that has resulted in a lack of protection over their culture, even in comparison to Native American culture. It will then analyze the current legal framework for protecting Indigenous cultural property, specifically intangible cultural heritage, both within the United States and globally. Informed by that analysis, this Note will present important components to include in a possible legal framework for protecting intangible cultural heritage for Native Hawaiians

A freeze substitution fixation-based gold enlarging technique for EM studies of endocytosed nanogold-labeled molecules

We have developed methods to locate individual ligands that can be used for electron microscopy studies of dynamic events during endocytosis and subsequent intracellular trafficking. The methods are based on enlargement of 1.4 nm Nanogold attached to an endocytosed ligand. Nanogold, a small label that does not induce misdirection of ligand–receptor complexes, is ideal for labeling ligands endocytosed by live cells, but is too small to be routinely located in cells by electron microscopy. Traditional pre-embedding enhancement protocols to enlarge Nanogold are not compatible with high pressure freezing/freeze substitution fixation (HPF/FSF), the most accurate method to preserve ultrastructure and dynamic events during trafficking. We have developed an improved enhancement procedure for chemically fixed samples that reduced auto-nucleation, and a new pre-embedding gold enlarging technique for HPF/FSF samples that preserved contrast and ultrastructure and can be used for high-resolution tomography. We evaluated our methods using labeled Fc as a ligand for the neonatal Fc receptor. Attachment of Nanogold to Fc did not interfere with receptor binding or uptake, and gold-labeled Fc could be specifically enlarged to allow identification in 2D projections and in tomograms. These methods should be broadly applicable to many endocytosis and transcytosis studies

Ligand Valency Affects Transcytosis, Recycling and Intracellular Trafficking Mediated by the Neonatal Fc Receptor

The neonatal Fc receptor (FcRn) transports IgG across epithelial cell barriers to provide maternal antibodies to offspring and serves as a protection receptor by rescuing endocytosed IgG and albumin from lysosomal degradation. Here we describe the generation of polarized Madin–Darby canine kidney (MDCK) cells expressing rat FcRn (rFcRn) to investigate the potential requirement for ligand bivalency in FcRn-mediated transport. The rFcRn-MDCK cells bind, internalize and bidirectionally transcytose the bivalent ligands IgG and Fc across polarized cell monolayers. However, they cannot be used to study FcRn-mediated transport of the monovalent ligand albumin, as we observe no specific binding, internalization or transcytosis of rat albumin. To address whether ligand bivalency is required for transport, the ability of rFcRn to transcytose and recycle wild-type Fc homodimers (wtFc; two FcRn-binding sites) and a heterodimeric Fc (hdFc; one FcRn-binding site) was compared. We show that ligand bivalency is not required for transcytosis or recycling, but that wtFc is transported more efficiently than hdFc, particularly at lower concentrations. We also demonstrate that hdFc and wtFc have different intracellular fates, with more hdFc than wtFc being trafficked to lysosomes and degraded, suggesting a role for avidity effects in FcRn-mediated IgG transport

Suppression of Puberty in Transgender Teens

The purpose of our study is to investigate the suppression of puberty for transgender teens. We will examine current and developing treatments, ethical limitations, and the overarching questions that cause hesitation in healthcare providers. Many of the issues faced by these children stem from their age, and thus their ability to make rational decisions for themselves. Proposals against treatment can create psychosocial developmental issues in teens and increase developing gender dysphoria; the psychological distress that results from the inability to align one’s assigned sex with their new gender identity. But fears of regretful decision-making leads to the question of what is more ethical, the postponement of treatment due to lack of self-awareness, or the alleviation of suffering with the risk of reversal

Comparison of colorimetric analyses to determine cortisol in human sweat

Colorimetric analysis, which relies on a chemical reaction to facilitate a change in visible color, is a great strategy for detecting cortisol, which is necessary to diagnose and manage the wide variety of diseases related to the hormone, because it is simple in design, inexpensive, and reliable as a standard cortisol analysis technique. In this study, four different colorimetric cortisol analyses that use various chromogens, which include sulfuric acid, Porter−Silber reagent, Prussian blue, and blue tetrazolium, are studied. Modifications to the classic Porter−Silber method are made by increasing the carbon content of the alcohol and adding gold nanoparticles, which result in a twofold increase in reaction rate and a slight decrease in the limit of detection (LoD). After a comparison of the reaction rate, LoD, dynamic range, characteristic peaks, and color stability of all methods, blue tetrazolium demonstrates a low LoD (97 ng/mL), broad dynamic range (0.05−2 μg/mL), and quick reaction rate (color development as fast as 10 min), which are well within the requirements for human biofluids. Cortisol in artificial saliva and sweat and in human sweat was determined while confirming that no excipients or other biomarkers interfered with the reactions. Twenty-one human sweat samples were tested using blue tetrazolium and revealed a significant difference between male and female apocrine cortisol concentrations and showed a highly significant difference between apocrine and eccrine cortisol concentrations. Colorimetric methods of cortisol can compete with existing electrochemical sensors because of their similar accuracy and detection range in certain wearable biosensor applications. The simplicity of colorimetric methods advances potential applications in skin-interfaced bio-electronics and point-of-care devices

Development and pilot of a low-literacy diabetes education book using social marketing techniques

Introduction: The primary objective of this work was to develop a diabetes education book, to pilot its use, and to evaluate its impact on patient care. The secondary objective was to compare the value of providing only the boo

Open-source software in an occupational health application: the case of Heales Medical Ltd

In this paper, we provide a case study of a small--company, Heales Medical Ltd., that has chosen to use opensource software to develop an online integrated patient management system. We use this case study to examine some preconceptions of open-source technology in the light our experiences of open source in the Heales project. We identify costs and cost savings as being the primary business considerations, and identify software purchase and licensing as key advantages for open-source. We then look at development issues including software evaluation, implementation and programming, and identify these as more costly for open-source, in terms of time and effort expended. We also look at issues to do with open source licensing, which our initial investigations suggest is confusing and requires--further analysis. Overall, the ability to modify the source code is regarded as a key benefit of open-source software, but in a business environment like Heales, this is of little importance