Search results for "electrophoresis"

Special Program of Differentiation Expressed in Keratinocytes of Human Haarscheiben: An Analysis of Individual Cytokeratin Polypeptides

1993

Human haarscheiben, epidermal Merkel cell-rich sensory organs of hairy skin, were studied for the expression of various cytokeratin (CK) polypeptides and other epithelial and neuronal differentiation markers by applying immunoperoxidase and immunofluorescence microscopy to frozen sections and by two-dimensional gel electrophoresis. The basal clusters of Merkel cells were specifically detected by antibodies against CK 20. Haarscheiben keratinocytes were unique mainly by the prominent expression of CK 17 in the lower and middle layers. Further differences as compared to keratinocytes of usual epidermis included the enlargement of the basal compartment, characterized by the expression of CK 5 …

Enzymatic modification of low-density lipoprotein in the arterial wall: a new role for plasmin and matrix metalloproteinases in atherogenesis.

2004

Objective— Functionally interactive proteases of the plasminogen/plasmin and the matrix metalloproteinase (MMP) system degrade and reorganize the extracellular matrix of the vessel wall in atherosclerosis. Here we investigated whether such proteases are able to confer atherogenic properties onto low density lipoprotein by nonoxidative modification. Methods and Results— Similar to the recently described enzymatically-modified low-density lipoprotein (E-LDL), native LDL exposed to plasmin or matrix MMP-2 or MMP-9 and cholesterylester-hydrolase (CEH) showed extensive deesterification, with ratios of free cholesterol to total cholesterol rising to 0.8 compared with 0.2 in native LDL. When the …

Identification of a Lipid Transfer Protein as a New Allergen From Morus alba Pollen.

2017

Blood levels of nitric oxide and DNA breaks assayed in whole blood and isolated peripheral blood mononucleated cells in patients with multiple sclero…

2019

Abstract Oxidative stress, especially overproduction of nitric oxide (NO), is considered to be one of the crucial factors in the pathogenesis of multifactorial multiple sclerosis (MS). DNA breaks could be one of the consequences of oxidative stress; however, data on DNA breakage in MS are very few and contradictory. There are no data on direct measurements of NO production in the blood of MS patients. The goal of this study was to determine the level of single-stranded DNA breaks in whole blood or isolated peripheral blood mononuclear cells (PBMNCs) by means of alkaline single cell gel electrophoresis (comet assay) and to evaluate production of NO in the human blood by applying electron par…

Determination of ethyl sulfate – a marker for recent ethanol consumption – in human urine by CE with indirect UV detection

2006

A CE method for the determination of the ethanol consumption marker ethyl sulfate (EtS) in human urine was developed. Analysis was performed in negative polarity mode with a background electrolyte composed of 15 mM maleic acid, 1 mM phthalic acid, and 0.05 mM cetyltrimethylammonium bromide (CTAB) at pH 2.5 and indirect UV detection at 220 nm (300 nm reference wavelength). This buffer system provided selective separation conditions for EtS and vinylsulfonic acid, employed as internal standard, from urine matrix components. Sample pretreatment of urine was minimized to a 1:5 dilution with water. The optimized CE method was validated in the range of 5-700 mg/L using seven lots of urine. Intra-…

C4A deficiency and nonresponse to hepatitis B vaccination

2002

Hepatitis B vaccination failure has been linked to the presence of certain human leukocyte antigen class II alleles. However, the functional background of these associations has remained unclear. Complement component C 4 is encoded within the major histocompatibility complex and is essential for classical pathway activation.Healthy individuals (n=4269) were vaccinated in a prospective trial with Engerix B. Nonresponse was classified as anti-HBs10 U/l after the last vaccination. Seventy-three nonresponders (NR) (1.7%) were identified. For comparison 53 responders (R) (anti-HBs10 IU/l) were drawn randomly from the same cohort. C4 allotyping was carried out by high-voltage agarose gel electrop…

Antibodies to Cathepsin G in Crohn's disease

1992

. Antibodies directed against antigens in human neutrophils have proved to be of great diagnostic value in certain systemic vasculitides. Recent reports have focused the attention on these antigens as targets of antibodies in sera of patients with inflammatory bowel disease. We investigated the sera drawn from 60 patients suffering from biopsy proven Crohn's disease and 15 patients with active ulcerative colitis. Using sensitive enzyme-linked immunosorbent assays with purified antigens and Western blotting the following antibodies could be demonstrated: cathepsin G (cat-G) antibodies IgG 38.3%, IgM 13.3%, IgA 23.3% and antibodies against human leucocyte elastase (HLE) IgG, IgA, IgM 3.3%. Lo…

Red cell enzyme polymorphisms in Bulgaria.

1972

7 human red cell enzyme polymophisms have been typed on a sample of n=138 unrelated adults from Bulgaria, which revealed the following gene frequencies: ADA1=0.8623. ADA2=0.1376; AK1=0.9637, AK2=0.0362; 6-PGDA=0.9891, 6-PGDC=0.0108; PGM11=0.8346, PGM12=0.1653; PA=0.1596, PB=0.7983, PC=0.0420. In the LDH-system one B-subunit variant was found, whilst no Peptidase A or B variant could be observed. The anthropological significance of these findings is discussed.

Comparability of two different polyacrylamide gel electrophoresis methods for the classification of LDL pattern type

2011

Abstract Background The measurement of small dense low-density lipoprotein (sdLDL) particles is relevant when assessing cardiovascular risk. However, there is as yet no referenced method for the determination of LDL subfractions or a standardized comparison of the methods currently available. Therefore, the aim of this study was to compare the pattern of LDL particles measured by polyacrylamide tube gel electrophoresis (PTGE) and polyacrylamide gradient gel electrophoresis (PGGE) and to correlate the results with triglyceride concentration. Materials and methods Serum samples were collected from 177 patients. Lipid profile and LDL particle size were assessed using PTGE and PGGE. Results Pea…

Characterization of lacrimal proline-rich protein 4 (PRR4) in human tear proteome

2014

This study was initiated considering the lack of comprehensive characteristics profile of PRR4 in tears of healthy subjects. Therefore, detailed characterizations of PRR4 from basal tears employing in-gel and in-solution digestions for MS systems are presented herein. First, pooled tear samples (n = 10) were utilized to identify PRR4-rich region/spots in 1DE/2DE gels employing LC-MALDI-MS and 1DE-LC-ESI-LTQ-Orbitrap-MS systems. PRR4-rich region and ten spots with vast polymorphisms (Mr : 17-30 kDa, pI: 3.0-6.6) were identified in 1DE and 2DE gels, respectively. In addition, combinations of four types of PTMs, which are methylation, acetylation, oxidation, and pyroglutamate formation, were i…