Assay Using Brain Homogenate for Measuring the Antioxidant Activity of Biological Fluids

1974 ◽  
Vol 47 (3) ◽  
pp. 215-222 ◽  
Author(s):  
J. Stocks ◽  
J. M. C. Gutteridge ◽  
Rosemary J. Sharp ◽  
T. L. Dormandy
Keyword(s):  
Antioxidant Activity ◽  
Cerebrospinal Fluid ◽  
Synovial Fluid ◽  
Biological Fluids ◽  
Brain Homogenate ◽  
Normal Serum

1. Fresh ox-brain homogenate prepared under standard conditions at 4°C and stored at −20°C autoxidizes spontaneously and reproducibly when re-heated to 37°C. 2. The preparation can be used for assaying the antioxidant activity of serum and of other biological fluids. The preparatory and assay procedures are described. 3. The antioxidant activity of normal serum, cerebrospinal fluid, synovial fluid and of various known antioxidants have been compared.

scholarly journals Oxidative damage to hyaluronate and glucose in synovial fluid during exercise of the inflamed rheumatoid joint. Detection of abnormal low-molecular-mass metabolites by proton-n.m.r. spectroscopy

1991 ◽  
Vol 273 (2) ◽  
pp. 459-467 ◽  
Author(s):  
M Grootveld ◽  
E B Henderson ◽  
A Farrell ◽  
D R Blake ◽  
H G Parkes ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Molecular Mass ◽  
Biological Fluids ◽  
Reactive Oxygen ◽  
Oxygen Radical ◽  
Normal Serum ◽  
Joint Disease ◽  
Serum Samples ◽  
Rheumatoid Synovial Fluid ◽  
Gamma Radiolysis

Proton Hahn spin-echo n.m.r. spectroscopy was employed to detect abnormal metabolites present in rheumatoid synovial fluid that are derived from the deleterious generation of reactive oxygen radical species during exercise of the inflamed rheumatoid joint. A resonance attributable to a low-molecular-mass N-acetylglucosamine-containing oligosaccharide formed by the oxygen-radical-mediated depolymerization of synovial-fluid hyaluronate was clearly demonstrable when subjects with inflammatory joint disease were exercised. Moreover, formate, which may be derived from the attack of OH.radical on synovial-fluid carbohydrates, was also readily detectable in these samples. gamma-Radiolysis of rheumatoid synovial fluid samples and aqueous solutions of hyaluronate also gave rise to the production of the low-molecular-mass hyaluronate-derived oligosaccharide species and markedly elevated concentrations of (non-protein-bound) formate in the biological fluids. As expected, corresponding spectra of gamma-irradiated blood serum samples obtained from normal volunteers did not contain the signal attributable to the low-molecular-mass oligosaccharide species, but the formate resonance (barely detectable in non-irradiated normal serum samples) became clearly visible. Additionally, a curious increase in the effective concentration of non-protein-bound low-molecular-mass metabolites such as acetate, citrate, lactate and glutamine was observed after gamma-radiolysis of all biological fluids studied. The hyaluronate-derived low-molecular-mass oligosaccharide species and formate are suggested as novel markers of reactive oxygen radical activity in the inflamed rheumatoid joint during exercise-induced hypoxic/reperfusion injury.

Solid-phase structures of cerebrospinal fluid in diagnosis of early asymptomatic neurosyphilis

2018 ◽  
pp. 56-61
Author(s):  
С.Н. Шатохина ◽  
Н.А. Кузнецова ◽  
В.Н. Шабалин
Keyword(s):  
Cerebrospinal Fluid ◽  
Visual Analysis ◽  
Solid Phase ◽  
Biological Fluids ◽  
Laboratory Methods ◽  
Phase Structures ◽  
Morphological Picture ◽  
Diagnostic Technology

Цель проведённого исследования состояла в оценке эффективности визуального анализа твёрдофазных структур спинномозговой жидкости для диагностики ранних форм нейросифилиса. Методы. Использован метод краевой дегидратации биологических жидкостей, входящий в состав авторской диагностической технологии «Литос-система». Диагностика раннего асимптомного нейросифилиса заключается в выявлении деструктивных образований в форме овалов в морфологической картине твёрдой фазы спинномозговой жидкости. Результаты. Проведён сравнительный анализ результатов исследования спинномозговой жидкости у 19 больных с подтверждённым диагнозом «ранний асимптомный нейросифилис», полученных традиционными лабораторными методами и методом краевой дегидратации. Выявлено, что локализация овалов внутри сферолитов указывает на длительность заболевания нейросифилисом менее трёх лет, а вне сферолитов - от трех до пяти лет. Заключение. Метод краевой дегидратации позволяет диагностировать ранний асимптомный нейросифилис по наличию деструктивных образований в форме овалов в морфологической картине твёрдой фазы спинномозговой жидкости. The aim of this study was to evaluate effectiveness of visual analysis of solid-phase structures in cerebrospinal fluid to diagnose early forms of neurosyphilis. Methods. We used a method of marginal dehydration of biological fluids as a part of the author’s diagnostic technology, Litos-System. Early asymptomatic neurosyphilis is diagnosed based on detection of destructive, oval-shaped formations in the morphological picture of cerebrospinal fluid solid phase. Results. Data from analyses of cerebrospinal fluid performed with traditional laboratory methods and the method of marginal dehydration were compared for 19 patients with documented diagnosis of early asymptomatic neurosyphilis. A localization of ovals within spherulites indicated a less than a three-year duration of neurosyphilis while a localization outside spherulites indicated a duration of three to five years. Conclusion. The method of marginal dehydration allows detecting early asymptomatic neurosyphilis based on the presence of destructive, oval-shaped formations in the morphological picture of cerebrospinal fluid solid phase.

scholarly journals Synovial Fluid Profile Dictates Nanopracticle Uptake into Cartilage- Implications of the Protein Corona for Novel Arthitis Treatments

2021 ◽  
Author(s):  
Ula von Mentzer ◽  
Tilia Selldén ◽  
LOISE Råberg ◽  
Gizem Erensoy ◽  
Anna-Karin Hultgård-Ekwall ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Drug Delivery ◽  
Synovial Fluid ◽  
Electrostatic Interactions ◽  
Fetal Calf Serum ◽  
Biological Fluids ◽  
Calf Serum ◽  
Protein Corona ◽  
Drug Delivery Vehicles ◽  
Drug Concentrations

<div>Intra-articular drug delivery strategies aiming to deliver drugs in diseases affected by cartilage-related issues are using electrostatic interactions to penetrate the dense cartilage matrix. This enables delivery of sufficient drug concentrations to the chondrocytes to mediate the desired therapeutic effect. As it is well known that size and charge of nanoparticles affects its interactions with the surrounding biological fluids, where proteins adsorb to the NP surface, resulting in a protein corona. There are, however, no studies investigating how the formed protein coronas affect cartilage uptake and subsequent cellular uptake, nor how they affect other cells present in the synovium of such diseases. Here, we explore the differences between the protein coronas that form when NP are incubated in synovial fluid from osteoarthritic and rheumatoid arthritis patients and compare this to results obtained using fetal calf serum (FCS), as guide for researchers working on joint drug delivery. We demonstrate that the protein corona indeed affects the uptake into cartilage, where there are major differences between the model proteins in fetal calf serum, as compared to synovial fluid from rheumatoid arthritis patients as well as osteoarthritis patients. The data suggests that when developing drug delivery vehicles for joint diseases that leverages electrostatic interactions and size, the interactions with proteins in the biological milieu is highly relevant to consider.</div>

DDRE-08. A BIOANALYTICAL LC-MS/MS ASSAY TO QUANTIFY TOTAL AND UNBOUND LY3214996, ABEMACICLIB AND ITS ACTIVE METABOLITES IN HUMAN PLASMA, CEREBROSPINAL FLUID AND HUMAN GLIOBLASTOMA

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi75-vi76
Author(s):  
Tigran Margaryan ◽  
Mackenna Elliott ◽  
Garry Hook ◽  
Nader Sanai ◽  
Artak Tovmasyan
Keyword(s):  
Cerebrospinal Fluid ◽  
Human Plasma ◽  
Equilibrium Dialysis ◽  
Brain Homogenate ◽  
Gradient Elution ◽  
Recurrent Glioblastoma ◽  
Active Metabolites ◽  
Unbound Fraction ◽  
Human Glioblastoma ◽  
Four Levels

Abstract BACKGROUND Here, we report on our development and validation of a sensitive and rapid LC-MS/MS method for the determination of total and unbound concentrations of ERK inhibitor LY3214996, CDK4/6 inhibitor abemaciclib and its M2 and M20 active metabolites in human plasma, cerebrospinal fluid and human glioblastoma tissue. METHODS Analytes were extracted from patient plasma, CSF and glioblastoma homogenate samples by protein precipitation with methanol. Four levels of quality controls were used during validation. The method was validated according to FDA guidelines and CAP/CLIA regulations. Equilibrium dialysis was performed to determine unbound fraction of four analytes in plasma and brain tissue. The detection was performed on Sciex QTRAP 6500+ mass spectrometer in positive electrospray ionization mode. RESULTS The method was validated over a concentration range from 0.2-500 nmol/L for all four analytes. The analytical separation was optimized on Phenomenex Kinetex™ F5 column with total run time of 3.8 min using gradient elution. For all the analytes the maximum coefficient of variation for intra- and inter-day precision was 12.0% and the accuracy was within 90.2-119.7% in all matrixes. The analytes are stable in plasma and brain homogenate for at least 19 hours and 6 hours at room temperature (RT), respectively. Stability of stock and working solutions was demonstrated for at least 15 hours (RT) and 28 days (2-8°C). The unbound fraction of the analytes in pooled human plasma and brain were determined to be 0.371 and 0.065 for LY3214996, 0.026 and 0.013 for abemaciclib, 0.052 and 0.008 for M2, 0.024 and 0.021 for M20, respectively. CONCLUSIONS A bioanalytical method to quantify LY3214996, abemaciclib and its M2 and M20 metabolites is successfully developed and validated. The method is currently applied to evaluate plasma pharmacokinetics and CNS penetration of LY3214996 and abemaciclib in recurrent glioblastoma patients in an ongoing Phase 0 clinical trial (NCT04391595).

Time-resolved immunofluorometric assay of complement C3: application to cerebrospinal fluid

1993 ◽  
Vol 39 (2) ◽  
pp. 309-312 ◽  
Author(s):  
O Gaillard ◽  
D Meillet ◽  
M C Diemert ◽  
L Musset ◽  
J Delattre ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Neurological Disorders ◽  
Biological Fluids ◽  
Clinical Applications ◽  
Complement C3 ◽  
Complement Components ◽  
Time Resolved ◽  
Sandwich Type ◽  
Analytical Range ◽  
Immunochemical Methods

Abstract Complement components have a role in various neurological disorders. Complement C3 can be measured by immunochemical methods, but only radioimmunoassays and electroimmunodiffusion assays (EIDs) are sufficiently sensitive to be applied to biological fluids in which the C3 concentration is low, especially cerebrospinal fluid (CSF). We report a sandwich-type time-resolved immunofluorometric assay (TR-IFMA) for C3 in CSF. The linearity (0.7-3650 micrograms/L) and intra- (CV &lt; 4.8%) and inter-assay (CV &lt; 10.9%) precision were satisfactory and the results agreed with those of EID. The assay is extremely sensitive (&lt; 1 microgram/L) and its analytical range is large and well suited to clinical applications. This simple TR-IFMA is thus a nonisotopic alternative to radioimmunoassay for the quantification of complement C3 in CSF.

Quantitative Nephelometric Microdetermination of Alpha-Globulins in Serum and Cerebrospinal Fluid

1956 ◽  
Vol 2 (3) ◽  
pp. 195-213 ◽  
Author(s):  
Abraham Saifer ◽  
Michael C Zymaris
Keyword(s):  
Cerebrospinal Fluid ◽  
Quantitative Determination ◽  
Salt Concentration ◽  
Protein Fraction ◽  
Specific Interaction ◽  
Normal Serum ◽  
Electrophoretic Method ◽  
Serum Fractions ◽  
Standard Serum

Abstract 1. A micronephelometric method is presented for the quantitative determination of total α-globulins in both serum and cerebrospinal fluid. The method is based on the relatively specific interaction of this protein fraction with the cationic detergent, Octab, at pH 6.65 in collidine-NaCl (.08M) buffer. 2. The various Cohn's protein fractions (Method 6) (25) were investigated with the nephelometric procedure. In comparison with normal serum, Fractions IV-6, IV-1, and IV-4, respectively showed the greatest detergent reactivity per milligram of α-globulin. 3. The variations of such factors as the concentrations of reagents, protein/detergent ratio, and salt concentration were reinvestigated. 4. Analysis of 14 normal and 33 pathologic sera gave excellent checks with the total α-globulin values obtained with the Tiselius electrophoretic method. 5. Application of the procedure to the analysis of the α-globulin content of cerebrospinal fluid required the addition of standard serum to 0.5-ml. aliquots of spinal fluid for stoichiometric results.

Potentiometric analysis for sodium and potassium in biological fluids.

1982 ◽  
Vol 28 (1) ◽  
pp. 170-172 ◽  
Author(s):  
E Langhoff ◽  
I Steiness
Keyword(s):  
Cerebrospinal Fluid ◽  
Biological Fluids ◽  
Flame Photometry ◽  
Aqueous Samples ◽  
Physiological Range ◽  
Relative Decrease ◽  
Potentiometric Analysis ◽  
Sample Water ◽  
Uremic Patients ◽  
Sodium And Potassium

Abstract Results obtained with a potentiometric analyzer, NOVA 1, specific for sodium and potassium, were compared with those by flame photometry. Both instruments showed linearity within a physiological range of sodium and potassium concentrations and had similar precisions. Volume displacements from addition of albumin or Intralipid to aqueous samples yielded the predicted lower flame-photometric results because of the relative decrease in sample water. There may be a small interaction between sodium and albumin. Physiological measurements on plasma from uremic patients showed no change after dialysis that could be ascribed to a decrease in interaction of these ions with creatinine and urea. Potentiometric values for sodium and potassium did not differ significantly, whether measured in cerebrospinal fluid or in the corresponding plasma. Results for urine were the same potentiometrically and by flame photometry.

scholarly journals Isolation and characterization of a membrane-attack-complex-inhibiting protein present in human serum and other biological fluids

1990 ◽  
Vol 265 (2) ◽  
pp. 471-477 ◽  
Author(s):  
M J Watts ◽  
J R Dankert ◽  
B P Morgan
Keyword(s):  
Cerebrospinal Fluid ◽  
Polyclonal Antibodies ◽  
Biological Fluids ◽  
Polyacrylamide Gel Electrophoresis ◽  
Membrane Attack Complex ◽  
Erythrocyte Membranes ◽  
Dependent Manner ◽  
Isolation And Characterization ◽  
Human Erythrocyte Membranes ◽  
Inhibiting Protein

We have previously reported the isolation of a membrane-attack-complex-inhibiting protein (MIP) from human erythrocyte membranes [Watts, Patel & Morgan (1987) Complement 4, 236] and the production of polyclonal antibodies to this protein. Here we report the identification in plasma, urine, saliva and cerebrospinal fluid of a protein immunochemically identical with the membrane-derived MIP. The protein has been isolated from plasma by immunoaffinity chromatography on an anti-(erythrocyte MIP)-Sepharose column and shown by SDS/polyacrylamide-gel electrophoresis to be of similar molecular mass to the erythrocyte protein (55 kDa non-reduced and 65 kDa under reducing conditions). Monoclonal antibodies have been raised against plasma MIP and used to establish a two-site enzyme-linked immunoadsorbent assay, enabling quantification of MIP in plasma, urine and cerebrospinal fluid. Plasma MIP, though not able to incorporate spontaneously into membranes, was deposited on heterologous and homologous erythrocyte membranes during complement activation in a C8-dependent manner. Depletion of MIP from plasma resulted in enhancement of the lytic capacity of the plasma on heterologous erythrocytes.

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