ABSTRACT
A 68-year-old female patient with multiple myeloma exhibited advanced nodular glomerulosclerosis. Immunofluorescence of the kidney showed kappa light chain deposition in the mesangium and in glomerular and tubular basement membrane. Isoelectric focusing and immunofixation of urinary proteins revealed an isolated kappa light chain with an unusually high isoelectric point of 8.4. Most light chain proteins have isoelectric points in the 4.6 to 6.7 range. Since loss of fixed negative charges may precede experimental glomerulosclerosis, it is proposed that this cationic circulating kappa chain may have interacted with glomerular polyanion, thereby inducing a nodular sclerotic reaction leading to irreversible renal damage.
Subject(s)
Glomerulonephritis/etiology , Glomerulosclerosis, Focal Segmental/etiology , Immunoglobulin kappa-Chains/analysis , Kidney Glomerulus/immunology , Multiple Myeloma/complications , Aged , Female , Fluorescent Antibody Technique , Glomerulosclerosis, Focal Segmental/immunology , Glomerulosclerosis, Focal Segmental/pathology , Humans , Immunoglobulin kappa-Chains/immunology , Isoelectric Focusing , Isoelectric Point , Kidney Glomerulus/pathology , Microscopy, Electron , Multiple Myeloma/immunology , Multiple Myeloma/pathologyABSTRACT
A topographic method for locating colorless proteins in focused Sephadex gel slabs is presented. The fluorescent isoelectric banding pattern of the carrier ampholyte (Servalyt) is utilized as a map to locate the proteins indirectly and non-destructively, and to guide their excision for preparative isolation. The method is illustrated with small-scale isolations of the major components of ovalbumin and human serum albumin, and with a scaled-up preparative isolation of the A and B components of beta-lactoglobulin. Fluorescence is used to localize accurately individual fractions of Servalyt in Sephadex slabs for preparative isolation. Isolated fractions were used to enrich selectively specific zones in the regular Servalyt mixture to increase the separation between close-lying proteins during focusing.
Subject(s)
Isoelectric Focusing/methods , Proteins/isolation & purification , Electrolytes , Humans , Lactoglobulins/isolation & purification , Ovalbumin/isolation & purification , Serum Albumin/isolation & purification , Ultraviolet Rays , Ampholyte MixturesABSTRACT
1. The isoelectric banding patterns of three commercial carrier ampholytes (pH range 2-11) were visualized in both Sephadex gel slabs and on paper prints, by means of ultraviolet light. 2. The fluorescence pattern of Servalyt is described in detail. This pattern, which has sharply delineated bands, is characteristic, and is reproducible from run to run with different batches of Servalyt. A total of 56 fluorescent bands were identified on gel slabs, and 62 on prints. 3. Methods are described for identifying individual fluorescent bands on prints. A classification scheme for organizing the banding pattern of Servalyt and naming individual bands is presented. 4. On gel slabs and on prints, the fluorescent pattern of a carrier ampholyte may be used analytically as a map for the location and identification of focused protein bands. The utility of the fluorescent pattern for characterising commercial ampholyte mixtures is discussed. 5. Individual fluorescent bands on the gel slab have characteristic pH values, which are constant from run to run and remain constant in the presence of focused sample proteins. On prints, these bands serve as permanent internal markers for assigning isoelectric points to protein bands.
Subject(s)
Buffers/analysis , Isoelectric Focusing/methods , Hydrogen-Ion Concentration , Spectrometry, Fluorescence , Spectrophotometry, Ultraviolet/methodsSubject(s)
Immunoglobulins/cerebrospinal fluid , Multiple Sclerosis/immunology , Female , Humans , Immunoglobulin A/cerebrospinal fluid , Immunoglobulin G/cerebrospinal fluid , Immunoglobulin M/cerebrospinal fluid , Male , Multiple Sclerosis/cerebrospinal fluid , Nerve Tissue Proteins/cerebrospinal fluid , Nervous System Diseases/cerebrospinal fluid , Nervous System Diseases/immunologyABSTRACT
A gas-chromatographic analysis for the succinimide anticonvulsant drugs--ethosuximide, methsuximide, and phensuximide--in 1.0 ml of serum was modified to improve its reliability, speed, and precision. A separate procedure for 10-100 mul of serum was also developed. Neither method requires an initial preparation of derivatives. The working range of each method is about 10-100 mg/liter for the macro-method, 2-100 mg/liter for the micro-method. The methsuximide metabolite, N-desmethylmethsuximide, is included in both methods. Concentrations of N-desmethylmethsuximide in the blood of two patients with petit mal epilepsy are reported.
Subject(s)
Ethosuximide/blood , Succinimides/blood , Animals , Chromatography, Gas/methods , Epilepsy, Absence/blood , Epilepsy, Absence/drug therapy , Ethosuximide/therapeutic use , Microchemistry , Succinimides/therapeutic use , TemperatureSubject(s)
Dopamine/analysis , Fluorescence , Indoles , Photochemistry , Spectrophotometry , Ultraviolet RaysABSTRACT
Various difficulties in the respirometry of filamentous fungi were avoided by using samples of moist mycelium layered thinly on tantalum grids. A variety of measurements is feasible with such preparations and are illustrated with samples of Schizophyllum commune from liquid and solid cultures.
