Capillary and Microchip Electrophoresis for the Monitoring of Disease Causing Amyloid Proteins

The detection of oligomers and aggregates formed by two amyloid proteins, insulin and amyloid-beta (AB), is of particular importance due to the role which these species play in Diabetes and Alzheimer\u27s disease, respectively. However, existing techniques are limited in the ability to detect insulin and AB; oligomers due to the fact that these early aggregates are transient, present at low concentrations, and difficult to isolate. Improvements must be made to existing techniques or alternative techniques must be explored in order to identify and quantify the size of these oligomeric and aggregate species without disrupting their structure. Capillary and microchip electrophoresis (CE and ME) are two promising electrophoretic methods for amyloid oligomer and aggregate detection. The present work demonstrated the potential for CE and ME to detect native aggregation of insulin and AB proteins, in particular the formation of oligomers and aggregates. Furthermore, the effect of hydrodynamic size on separation was increased through the use of a highly entangled polymer matrix introduced into the capillary, thus offering the potential to resolve populations of amyloid species. Using this novel separation, we investigated variables such as sample salt concentration, sample preparation method, and fluorescent dye structure. We demonstrated the ability of CE with ultraviolet detection (UV-CE) to detect native insulin aggregates estimated to range in size from 30 - 100 kDa and native AB1-40 aggregates estimated to range in size from 10 - 30 kDa, 10 - 300 kDa and \u3e 300 kDa. These studies demonstrated the successful detection of physiological concentrations (pM) of monomeric fluorescein isothiocyanate labeled insulin (FITC-insulin) and carboxy- fluorescein labeled AB1-42 (FAM-AB1-42) using CE with laser induced fluorescence detection (LIF-CE). However, the detection of oligomeric and aggregate species was altered from unlabeled samples, indicating the need for future work in the study of fluorescent dye attachment

Understandings of Education in an African Village: The Impact of Information and Communication Technologies

Teaching/Communication/Extension/Profession,

Unveiling the Mysteries of Aceh, Indonesia: Local and Global Intersections of Women\u27s Agency

Forces of globalization, local culture, and Islam continuously inform one another and dynamically manifest in cultures across the world. Scholars often assume that these influences may have distinct and independent effects. However, we argue that these global forces occur simultaneously and they may contradict or complement each other along a spectrum within Aceh, Indonesia. The manifestations and responses vary depending on the nature of the interactions of global and local factors. This spectrum represents various ways in which women negotiate identity and agency, specifically within the context of the implementation of Shari’ah Law. This research investigates the specific ways in which women’s identities influence and are influenced by the globalization of feminism, matrifocal traditions, and Islamic veiling practices in Aceh. In the summer of 2012, the authors conducted field research in Aceh, Indonesia through interviews and observations. These included over 70 participants and 20 organizations which varied in formality and size. The interview participants include: activists, academicians, spiritual leaders, government officials, law enforcement agents, university students, and other community members from both rural and urban areas

Interkingdom Gene Transfer of a Hybrid NPS/PKS from Bacteria to Filamentous Ascomycota

Nonribosomal peptides (NRPs) and polyketides (PKs) are ecologically important secondary metabolites produced by bacteria and fungi using multidomain enzymes called nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs), respectively. Previous phylogenetic analyses of fungal NRPSs and PKSs have suggested that a few of these genes were acquired by fungi via horizontal gene transfer (HGT) from bacteria, including a hybrid NPS/PKS found in Cochliobolus heterostrophus (Dothideomycetes, Ascomycota). Here, we identify this hybrid gene in fungi representing two additional classes of Ascomycota (Aspergillus spp., Microsporum canis, Arthroderma spp., and Trichophyton spp., Eurotiomycetes; Chaetomium spp. and Metarhizium spp., Sordariomycetes) and use phylogenetic analyses of the most highly conserved domains from NRPSs (adenylation (A) domain) and PKSs (ketoacyl synthase (KS) domain) to examine the hypothesis that the hybrid NPS7/PKS24 was acquired by fungi from bacteria via HGT relatively early in the evolution of the Pezizomycotina. Our results reveal a unique ancestry of the A domain and KS domain in the hybrid gene relative to known fungal NRPSs and PKSs, provide strong evidence for HGT of the hybrid gene from a putative bacterial donor in the Burkholderiales, and suggest the HGT event occurred early in the evolution of the filamentous Ascomycota

Chronicles of Oklahoma

Article attempts to chronicle the ancestry of Nathaniel Pryor through documents surrounding his relatives and friends. Pryor took part in the Louis and Clark expedition to map the continental United States

Retinal detachment post-phacoemulsification in Bichons Frises: A retrospective study of 54 dogs

Objective—To compare rates of retinal detachment (RD) post phacoemulsification in American Bichons Frises with and without prophylactic retinopexy. Procedures—Medical records of 54 Bichons Frises undergoing phacoemulsification with or without prophylactic retinopexy between 2003–2013 in one or both eyes were reviewed from five Midwestern university veterinary teaching hospitals. Inclusion criteria were pre-operative ERG, at least 6 months of follow up post phacoemulsification, and absence of pre-existing RD as determined by ophthalmic examination and/or ultrasound. Statistical analyses used chi-squared and Wilcoxon rank-sum tests and Wilson confidence intervals with the p value Results—Phacoemulsification was performed without retinopexy in 79 eyes (42 dogs, non-PR group) and with prophylactic retinopexy in 23 eyes (12 dogs, PR group). Incidence of diabetes mellitus was 10/42 and 3/12 in the non-PR and the PR groups respectively (p=0.93). Intraocular lens implantation was performed in 40/42 non-PR dogs and 11/12 PR dogs, (p=0.63, 73/79 versus 21/23 eyes). At final re-examination, RD occurred in 4/79 eyes without retinopexy, compared to 0/23 RD in the retinopexy group. There was no statistically significant difference in RD rates between the two groups (p=0.27) Conclusions—These data provide no statistical evidence to support prophylactic retinopexy in Bichon Frises. Due to the low rate of retinal detachment following phacoemulsification without prophylactic retinopexy, the procedure appears to offer limited benefit to offset cost, procedural risk and risk of extended or repeated anesthesia in Bichons Frises

The Rhodococcus equi virulence protein VapA disrupts endolysosome function and stimulates lysosome biogenesis

Rhodococcus equi (R. equi) is an important pulmonary pathogen in foals that often leads to the death of the horse. The bacterium harbors a virulence plasmid that encodes numerous virulence-associated proteins (Vaps) including VapA that is essential for intracellular survival inside macrophages. However, little is known about the precise function of VapA. Here, we demonstrate that VapA causes perturbation to late endocytic organelles with swollen endolysosome organelles having reduced Cathepsin B activity and an accumulation of LBPA, LC3 and Rab7. The data are indicative of a loss of endolysosomal function, which leads cells to upregulate lysosome biogenesis to compensate for the loss of functional endolysosomes. Although there is a high degree of homology of the core region of VapA to other Vap proteins, only the highly conserved core region of VapA, and not VapD of VapG, gives the observed effects on endolysosomes. This is the first demonstration of how VapA works and implies that VapA aids R. equi survival by reducing the impact of lysosomes on phagocytosed bacteria

920-52 Are Provider Profiles Affected by Risk-adjustment Methodology? Results from the Cooperative Cardiovascular Project

Health care payors and consumers have a growing interest in risk-adjusted provider profiles. Using chart-abstracted clinical data from the Cooperative Cardiovascular Project, we ranked 28 hospitals performing bypass surgery in Alabama and Iowa by their risk-adjusted surgical mortality rates using three published risk-adjustment methodologies: Parsonnet (PI, O’Connor (a) and Hannan (H). In total. 3653 bypass surgery cases performed from 6/92 to 3/93 were reviewed (mean 130 cases/hospital). The discriminatory abilities of each method for predicting surgical mortality were quite similar (area under ROC curves 0.72–0.75). Below, we display the risk-adjusted hospital rankings (comparing observed with expected mortality) by these three riskadjustment techniques:In terms of hospital rankings, there was generally close correlation between any two of the methods (Spearman's R=0.87,0.88, and 0.93, comparing P-O, P-H, and H-O). Rankings for an individual hospital varied, however, an average of ±3.3 ranks (range 0–12 ranks) depending on which riskadjustment methodology was used.ConclusionIn general. published methods of risk-adjustment for bypass surgery accurately identify institutions with low, moderate and high adjusted mortality outcomes. The precise ranking of an individual hospital. however, may vary depending on the risk adjustment method applied

Impact of nasogastric tubes on swallowing physiology in older, healthy subjects: a randomized controlled crossover trial

Background & aims: The presence of a nasogastric tube (NGT) affects swallowing physiology but not function in healthy young adults. The swallowing mechanism changes with increasing age, therefore the impact of a NGT on swallowing in elderly individuals is likely to be different but is not yet known. The aims of this study were to determine the effects of NGTs of different diameter on (1) airway penetration-aspiration, (2) pharyngeal residue, and (3) pharyngeal transit, in older healthy subjects