The different electrophoretic forms of post gamma-globulin their antigenic identity and their structural variability.

Post gamma-globulin, first described as a constant component of protein of cerebrospinal fluid and urine from patients with tubular disorders has also been found in other biological fluids. Post gamma-globulin from a single individual always migrated as several bands after storage. Three electrophoretic forms, immunochemically identical, have been isolated by gel chromatography, preparative continuous flow electrophoresis and ion-exchange chromatography. Their molecular weight was found to be approximately between 11 000 to 12 000. No difference between the three forms could be detected. The N-terminal amino acids were found to be Lys, Arg and Leu respectively for the three forms of post gamma-globulin. The "slow" and "fast" forms of post gamma-globulin seemed to differ by elimination of small basic peptides or amino acids from the N-terminal end of the protein. No enzymatic activity of post gamma-globulin was found, but this requires further investigations.

Quantitative estimation of cerebrospinal fluid globulins in multiple sclerosis. A comparison of electrophoretic and immunologic methods.

CSF samples were obtained from 44 multiple sclerosis patients during exacerbation and from 50 age and sex-matched non-multiple sclerosis controls. In an attempt ot select the most suitable test for diagnosis of multiple sclerosis, each sample was analyzed by three methods: (1) electrophoresis, (2) electroimmunodiffusion, and (3) radial immunodiffusion (RID). Electrophoresis, electroimmunodiffusion, and radial immunodiffusion yielded positive results in 50, 55, and 59 percent of the multiple sclerosis patients respectively. Electrophoresis, electroimmunodiffusion, and radial immunodiffusion yielded 10, 12, and 8 percent false positives (beyond mean +/- 1 S.D.) respectively. Thus the three tests did not differ much with regard to yield of true or false positive results. When factors such as simplicity, time and equipment involved, and the interval between the test and results are taken into account, radial immunodiffusion appears to be most suitable for an avverage CSF diagnostic laboratory. For a laboratory with special interest in multiple sclerosis, agarose gel-electrophoresis of concentrated CSF appears to be the test of choice.

Post-gamma globulin : tissue distribution and physicochemical characteristics of dog post-gamma globulin.

An animal model (dog) was established to investigate physicochemical and biological parameters of a basic low molecular mass protein---post-gamma globulin (p-gamma). In this study the distribution of this 11 000 daltons (11 kdaltons) protein was determined in extracts of the number of tissues. Extracts of parotid gland, brain and kidney contained high amounts of "free" p-gamma globulin (11 kdaltons). Two higher molecular mass forms (34k and 27 kdaltons) of this protein were demonstrated in the kidney and brain extracts. Extracts of ovary and uterus were devoid of the 27 k moiety and kidney extracts lacked the "free" p-gamma globulin (11k). These studies indicate that p-gamma globulin may be a part of a much larger molecule or that p-gamma globulin may be associated with another yet unknown protein. These findings may be important for clinical and pathological studies in human kidney and central nervous system (CNS) diseases.

Post-gamma globulin. II. Radioimmunoassay determination of levels of post-gamma globulin and beta 2-microglobulin.

Post-gamma globulin previously isolated and partially sequenced in this laboratory was used for production of polyclonal and monoclonal (hybridoma) antibodies. A radioimmunoassay method was developed for quantitation of post-gamma globulin with either antibody. The titration curves obtained were treated statistically and found practically indistinguishable. The sensitivity of the method adopted for the quantitation of post-gamma globulin in a variety of biological fluids was 0.13 ng/ml and the upper limit of precision was 2.5 ng/ml. The following results were obtained (mean +/- 1 SD): normal sera, 0.96 +/- 0.20 microgram/ml; pregnancy sera, 1.08 +/- 0.28 micrograms/ml; cord blood 2.08 +/- 0.33 micrograms/ml; hospitalized patient's sera, 1.3 +/- 0.54 micrograms/ml; geriatric subjects' sera 2.26 +/- 1.10 micrograms/ml; cerebrospinal fluid, 5.37 +/- 3.36 micrograms/ml; saliva, 1.22 +/- 0.67 micrograms/ml; synovial fluid, 1.27 +/- 0.41 micrograms/ml and urine, 0.11 +/- 0.125 microgram/ml. To shed light on the catabolism of post-gamma globulin the levels of beta 2-microglobulin were also measured radiometrically. Correlative statistical analysis of all the data have shown that renal handling of post-gamma globulin and beta 2-microglobulin may be very similar but not necessarily identical.

Serum time course of two brain-specific proteins, alpha(1) brain globulin and neuron-specific enolase, in tick-born encephalitis and Lyme disease.

METHODS: Time courses of the serum concentrations of two brain-specific proteins (BSP), alpha(1) brain globulin (alpha(1)BG, an astroglial marker) and neuron-specific enolase (NSE), were studied in patients with severe tick-born encephalitis (TBE) and Lyme disease (LD; neuroborreliosis). The concentrations were determined on the second day of the acute phase and then on the 7th, 12th, 18th, and 23rd days. Apparent rate constants for the elimination of the BSP from blood (k(e)) were calculated with the non-linear regression. RESULTS: In patients with TBE, the highest serum concentrations of alpha(1)BG and NSE, observed on the second day, were followed by their monotonic decrease to the normal levels reached by the 23rd day. The mean k(e) values for alpha(1)BG and NSE were found to be significantly different (0.086+/-0.003 vs. 0.057+/-0.006 day(-1), respectively; p<0.05). Higher serum levels of both BSP were observed in the more severe clinical cases and in the cases with unfavorable outcomes. Similar profiles were also observed for the serum alpha(1)BG and NSE in LD. CONCLUSIONS: These results suggest that, in the patients examined, the blood-brain barrier was partially impaired; the quantitative parameters of the serum BSP time courses can be indicative of the extents of the neuronal and/or glial lesions.

Cerebrospinal fluid composition of cattle with endotoxin-induced mastitis treated with isotonic (0.9%) or hypertonic (7.5%) sodium chloride.

This study examined the safety of intravenous hypertonic saline in cattle with experimental gram-negative endotoxemia. Cerebrospinal fluid (CSF) composition was examined in five control cows and eight treated cows 24 hours after the intramammary infusion of 1 mg of endotoxin. Four of the endotoxin challenged cows were treated intravenously with isotonic (0.9%) sodium chloride and four cows were treated intravenously with hypertonic (7.5%) sodium chloride. Decreased CSF osmolality, and sodium and alpha globulin concentrations and increased CSF concentrations of beta globulin were observed in both endotoxin-challenged saline-treated groups. No CSF compositional differences were observed between endotoxin-challenged cows receiving isotonic or hypertonic saline. Although no cytologic or biochemical evidence of salt poisoning was observed in cows receiving hypertonic saline, significant changes were observed in the CSF composition of both endotoxin-infused saline-treated groups.

Total protein, albumin quota, and electrophoretic patterns in cerebrospinal fluid of dogs with central nervous system disorders.

Cerebrospinal fluid was analyzed for total protein, albumin quota, and electrophoretic patterns of albumin and alpha-, beta-, and gamma-globulins in 10 healthy dogs and 35 dogs with CNS disorders. Values obtained in healthy dogs were used to establish control data. Samples also were collected from dogs with neurologic diseases that were classified according to clinical and pathologic diagnosis as inflammation, neoplasm, and spinal cord compression. The albumin quota and total CSF albumin values were used as indicators of blood-brain barrier disturbance. Four patterns were observed on agarose electrophoresis that included intrathecal immunoglobulin production, intrathecal immunoglobulin production combined with blood-brain barrier disturbance, blood-brain barrier disturbance, and unaltered CSF. These patterns correlated with the observed clinical and pathologic conditions. Seemingly, agarose electrophoresis of CSF is a simple and reliable technique that aids in the diagnosis of CNS disorders of dogs.

[Cerebrospinal fluid proteins: II. Normal values of protein fractions obtained by electrophoresis (variations related to race, sex and age)]. / Proteínas do líquido cefalorraqueano: II. Valores normais das frações proteicas obtidas por electroforese (variações ligadas a cor, sexo e idade).

The total protein content and protein fractions, obtained by electrophoresis on cellulose acetate strips, of CSF collected from the cisterna magna of 213 patients (with no neurological diseases) were determined in order to verify variations related to race, sex and age, as well as to establish the proteinogram normal limits. No differences between caucasians and coloured persons were observed with respect to CSF total protein and protein fractions. Children (5 months to 11 years old) do not present differences related to sex or age, on their proteinogram. Children's CSF total protein, relative values of pre-albumin, alpha 1--and beta-globulins, absolute values of albumin, alfa 2--, beta--, tau --and gamma-globulins, differ from those found in adults. Differences between males and females in the normal CSF proteinogram were found in adults. As a consequence of these findings, the CSF proteinogram normal limits for children, males and females were separately established. In adults, statistically significant positive correlation between age and the 7 protein fractions when expressed in mg/100 ml were observed, as well as between age and total protein. Comparison of the results obtained in this research with those found in some publications was carried out and is briefly discussed.

[Vogt-Koyanagi-Harada syndrome: report of 2 cases]. / Síndrome de Vogt-Koyanagi-Harada: relato de dois casos.

The Vogt-Kayanagi-Harada ssyndrome in two negro female patients with 37 and 40 year-old, respectively, are reported. The clinic, epidemiologic, heredo-familial and immunologic aspects are discussed.

Electrophoresis of normal lumbar cerebrospinal fluid proteins in the African.

Lumbar cerebrospinal fluid obtained from Zambian patients suspected of suffering from central nervous system conditions was concentrated over a hundred-fold and subjected to small-scale cellulose acetate electrophoresis (Millipore). The percentage of each protein fraction was determined by scanning. Only twenty samples were considered normal out of a total of 150 studies. These normal values were not statistically different from those of published data on Europeans and North American Caucasians. The high serum gamma globulin in African subjects is not reflected in the cerebrospinal fluid.

[Cerebrospinal fluid proteins in the course of subacute sclerosing panencephalitis in children]. / Bialka plynu mózgowo-rdzeniowego w przebiegu podostrego stwardniajacego zapalenia mózgu u dzieci.

In cerebrospinal fluid samples obtained from lumbar tap from 31 children with subacute sclerosing panencephalitis the concentration of total protein was determined and electrophoretic separation of protein fractions on paper was carried out from 1 to 28 months after the onset of first clinical symptoms. Moreover, in 22 children the concentration of IgG was determined by electroimmunodiffusion. The reference group included 22 children without organic diseases of the central nervous system. In nearly half the samples of the cerebrospinal fluid obtained from these children a rise was observed in total protein level, and in most samples a significant fall of albumin proportion and an evident rise of this relative proportion of globulins (tau + gamma) and IgG immunoglobulins was observed. No correlation was demonstrated between changes in cerebrospinal proteins and the phase of the disease and its duration.

[Evaluation of cerebrospinal fluid proteins in patients with subacute sclerosing panencephalitis (SSPE)]. / Ocena bialek plynu mózgowo-rdzeniowego u chorych na podostre stwardniejace zapalenie mózgu (SSPE).

CSF proteins of 62 patients with SSPE aged 6 to 26 years were analysed by polyacrylamide gel electrophoresis. The total IgG concentration in the CSF was increased to 16.8 +/- 10 mg/dl and the IgG index was raised to from 2.7 +/- 0.5 to 3.6 +/- 3.0. All SSPE patients had pathological subfractions gamma 4 and gamma 5. The per cent proportion of the gamma 1 subfraction was raised only in the youngest patients. A tendency was observed for higher values of the prealbumin:albumin index and the prealbumin: transferrin index. The youngest SSPE patients had a tendency for greater total protein concentration, lower prealbumin concentration, and highest IgG index values (3.6 +/- 3.0). In early disease period a tendency was noted for higher concentration of total protein, higher total IgG level and higher IgG index. These results suggest that SSPE patients have a constant stimulation of B-cells and variable increase of blood-brain barrier permeability.

[Significance of oligoclonal proteins and magnetic resonance imaging in the diagnosis of multiple sclerosis]. / Znaczenie bialek oligoklonalnych i magnetycznego rezonansu jadrowego w diagnostyce stwardnienia rozsianego.

The new techniques identifying inactive clinical injury, as well as revealing immunological disturbances connected with the CNS, are very helpful for arriving at the correct diagnosis of multiple sclerosis. In their analysis the authors of the work include six cases and emphasize the great practical significance of magnetic resonance imaging and oligoclonal bands in the cerebrospinal fluid in making the diagnosis of multiple sclerosis.

[Composition of cerebrospinal fluid protein in children with nervous system diseases according to disc-electrophoresis in polyacrylamide gel]. / Osobennosti sostava belkov spinnomozgovoi zhidkosti pri zabolevaniiakh nervnoi sistemy u detei po dannym disk-élektroforeza v poliakrilamidnom gele

The author studied the fractional content of proteins in the CSF in 105 children with different disorders of the nervous system and 35 children of a control group (aged 2-13 years) with the aid of disc-electrophoresis in polyacrylamide gel. The proteins of the CSF were studied in encephalitis, polyradiculoneuritis, cerebral arachnoiditis, brain tumors and epilepsy. The content of total proteins in the CSF in 42% of the studied children did not differ significantly from the usual age norm. More distinct changes were found in proteinogramms of the CSF where 14 protein fractions were singled out. It was established that a change in the content of separate fractions, their intercorrelation depended on the character of the pathological process.

High performance collection of cerebrospinal fluid in rats: evaluation of erythropoietin penetration after osmotic opening of the blood-brain barrier.

An important issue to be considered when studying a new drug for treatment of central nervous system (CNS) diseases is its ability to cross the blood-brain barrier (BBB) and distribute throughout the brain. As cerebrospinal fluid (CSF) has demonstrated to be an invaluable reservoir to study CNS availability of therapeutic proteins, we have developed an improved method for CSF sampling from the cisterna magna of rats. The technique enables the simple and rapid collection of adequate quantities (50-75 µl) of blood-free CSF, rendering a high percentage of animal survival (99%) without clinic or neurological consequences. Its success in avoiding blood contamination of CSF lays in the use of a mixture of lidocaine/ephinephrine topically injected in the rat's suboccipital area and neck. Another relevant feature of the methodology is its low cost, since the puncture device can be easily assembled with cheap and available materials and, more importantly, neither expensive stereotaxic equipment nor frame is required. The present method is demonstrated by studying the CSF pharmacokinetics of recombinant human erythropoietin (rhEPO), a well-studied therapeutic candidate for neurological diseases. Moreover, we applied this technique to evaluate a strategy of osmotic disruption of the BBB to achieve a faster delivery of rhEPO into the CNS.