Antigenic streptococcal components in acute glomerulonephritis.

Fluorescein-labeled immunoglobulin G fractions from serums of patients with acute glomerulonephritis and from many normal serums stained the glomerular basement membrane and mesangium of renal tissue from patients with early acute glomerulonephritis; these serums did not stain the corresponding tissues from patients with any other kidney disease. Previous absorption of the serum fraction with frozen and thawed nephritogenic beta hemolytic streptococci abolished all staining. Other bacteria studied did not abolish the staining. Only the plasma membrane of the streptococcus absorbed the immunoglobulin G fraction; such absorption eliminated staining. Fluorescein-labeled antiserums against streptococcal plasma membrane had staining properties similar to patients' serums.

Partial purification and properties of a plasminogen activator from human erythrocytes.

The lysis time of euglobulin clots made with whole blood (plasma and red cells) was very much shorter than that of clots made with plasma alone, indicating a fibrinolytic component in red cells. A plasminogen activator was found in the stroma-free hemolysate, and proteolytic activity was found in the stromal fraction. The plasminogen activator, purified by using diethylaminoethyl-cellulose (DEAE-cellulose) in a batch procedure followed by column chromatography, was called erythrokinase (EK). On preliminary characterization, EK appears to activate human and bovine plasminogen in a manner similar to urokinase (UK), as determined by fibrinolytic and caseinolytic assays. The two enzymes can be separated by DEAE chromatography and acrylamide-gel electrophoresis, however, and they hydrolyze acetyl-L-lysine methyl ester and benzoyl arginine methyl ester at different rates.

Partial characterization of antigenic streptococcal plasma membrane components in acute glomerulonephritis.

Fluorescein-labeled immunoglobulin G (IgG) fractions of serum from patients with acute poststreptococcal glomerulonephritis stained parts of the glomerular basement membrane and mesangium of kidney tissue obtained from the same patients during the early phase of the disease. Renal tissue obtained from normal individuals and from patients with other kidney diseases failed to stain with these IgG fractions. Preabsorption of the serum fractions with various freezethawed bacteria demonstrated that only certain group A streptococci abolished the staining capacity. Fractionation of the streptococci into cellular constituents indicated that it was predominantly the plasma membrane fraction which blocked the immune staining. Spectrofluorometry using alkali-solubilized renal tissue confirmed these observations in a quantitative manner. By sucrose density-gradient ultracentrifugation of the plasma membrane two possible antigens were isolated. One was soluble in phosphate-buffered saline and the other was insoluble. The soluble component was a lipoprotein with a molecular weight of approximately 120,000.

Relationship of nervous tissue transketolase to the neuropathy in chronic uremia.

Patients with chronic uremia develop neurologic defects which are similar to the demyelinating lesions seen in thiamine deficiency. The present study describes inhibitory effects of uremic material on nervous tissue transketolase, a thiamine-dependent enzyme of the pentose phosphate pathway which has been reported to have functional importance in the metabolism of myelinated nervous structures. Transketolase activity (TKA) of normal human brain and spinal cord was measured by the conversion of ribose-5-phosphate (R5P) to sedoheptulose-7-phosphate (S7P). TKA was significantly inhibited by plasma, cerebrospinal fluid and low molecular weight dialysate fractions obtained from patients with uremic neuropathy, but not by samples from normal subjects. The specific effect on transketolase by uremic material was established by showing suppressed formation of S7P from R5P also in the presence of excess cofactor thiamine pyrophosphate and of the other substrate xylulose-5-phosphate. Uremic plasma likewise inhibited a partially purified transketolase preparation from bakers' yeast.31 of 35 chronic uremic patients with inhibition values between 10 and 84% before or during the early phase of intermittent hemodialysis had evidence of neuropathy. Data of clinical grading of the neurologic deficits and values of motor nerve conduction velocity revealed a correlation between the extent of uremic neuropathy and the degree of nervous tissue transketolase inhibition. Hemodialysis markedly reduced the inhibitory effects of the patients' plasma and the data indicate that uremic patients who received effective long-term dialysis treatment show a parallel decline of transketolase inhibition and uremic neuropathy.The findings demonstrate that in patients with chronic renal failure, low molecular weight factors accumulate and inhibit nervous tissue transketolase. This biochemical defect-uncorrectable by thiamine but reversible by dialysis-may interfere with the metabolism of myelin-supporting cells, and/or of the axonal metabolism of medullated structures, and may thus contribute to the degeneration of myelinated nerves seen with uremic neuropathy.

Ultrastructural differentiation of glandular stomach (proventriculus) in chick embryo.

Cytochemical characterization of mucosubstances of chick glanular stomach (proventriculus) changes from 15 days of development to postnatal and adult stages was studied. To corroborate these data cytochemical, ultrastructural and ultracytochemical study of chick embryo proventriculus from 7 to 20 days of development was performed. At the 7th day several layers of undifferentiated cells formed an epithelium which covered the walls of the glandular stomach. Mocosubstances were not found. Between the 9th and 5th day a single layer of cylindrical cells was encountered forming invaginations which originated deep glands. Three types of cells were separated from the above mentioned layer, dark, clear and undifferentiated. The dark cells had organelles which are involved in protein synthesis and the clear ones were rich in mitochondria. Argentaffine cells appeared at 15th day instead mucosubstances formed a thin coat on the epithelium at 9th day which increased at the end of development in the apical cytoplasm and gland cells. These observations demonstrate that proventriculus of chick embryo has ultrastructurally differentiated cells involved with enzymatic and hydrochloric acid secretion after the 9th day. These progressive events are correlated with the digestion process of yolk during embryogenesis. At the end of development the proventriculus has completely organized the glandular layer.

Lachnellins A, B, C, D, and naphthalene-1,3,8-triol, biologically active compounds from a Lachnellula species (ascomycetes).

In the course of our search for new biologically active metabolites, lachnellin A (1), a metabolite with high cytotoxic and antimicrobial activities, the structurally related lachnellins B, C and D (3, 4, 7), and naphthalene-1,3,8-triol (8), an inhibitor of malate synthase (EC 4.1.3.2), were isolated from submerged cultures of the ascomycete Lachnellula sp. A 32-89. The antimicrobial, cytotoxic and phytotoxic activities of lachnellin A depended on its reactivity and could be abolished by the addition of cysteine. The enzyme inhibiting activity of (8) was due to reactive intermediates during melanization and was no longer observed in the presence of serum albumin. In addition, rac-scytalone (9), (+)-trans-3,4-dihydro-3,4,8-trihydroxy-1 (2H)-naphthalenone (10), 2,5-dihydroxytoluene (11), and (R)-(-)-5-methylmellein (12) were obtained from the same source and biologically characterized.

Detection of hepatitis B surface antigen in potentially contaminated human plasma and plasma fractions.

A new method is described for the quantitative detection of HBsAg in whole human plasma and in plasma fractions. The nonantigen proteins are digested with pepsin at low pH, and the antigen is precipitated with PEG. With use of only 20 ml of contaminated plasma, as few as 5.0 x 10(6) HBsAg particles/ml can be detected--a 40-fold increase in the apparent level of sensitivity of the Ausria II RIA (2.0 x 10(8) particles/ml). With 500 500 ml or more of plasma or plasma fractions, fewer than 5.0 x 10(5) particles/ml can be assayed--a 400-fold increase in RIA sensitivity and 1/10 the antigen concentration found in sera that proved infective when injected into chimpanzees. The pepsin-PEG method was used to quantitate the particles per milliliter in four equivocal RIA samples from the NIH Bureau of Biologics, three of which were definitely shown to contain antigen. The method has also been employed to detect fewer than 2.0 x 1010(8) particles/ml of HBsAg in deliberately contaminated high purity AHF concentrates and may be useful for monitoring plasma fractions prepared on a large scale or for detecting the antigen in equivocal samples from blood banks.

Tyromycin A: a novel inhibitor of leucine and cysteine aminopeptidases from Tyromyces lacteus.

Tyromycin A, a new inhibitor of leucine and cysteine aminopeptidases has been isolated from mycelial cultures of the basidiomycete Tyromyces lacteus (Fr.) Murr. Its structure was established as 1,16-bis-[4-methyl-2,5-dioxo-3-furyl]hexadecane (1) by spectral analysis and transformation into ester and imide derivatives.

Freedom from transmission of hepatitis-B of gamma-globulin and heat-inactivated plasma protein fraction prepared from contaminated human plasma by fractionation with solid-phase polyelectrolytes.

Plasma contaminated with hepatitis B surface antigen (HBsAg) and shown by others to be infectious when injected in a dilution of 1:1,000,000 in chimpanzees, was fractionated by a solid-phase polyelectrolyte (PE) procedure for its content of plasma protein fraction (PPF) and gamma-globulin (immune serum globulin; ISG). Quantitative Ausria II radioimmunoassays showed that nearly half the HBsAg was bound by the PE and could be eluted at low pH, while the rest was found in the heat-inactivated PPF. When the ISG was concentrated to 16%, the 13 mg/kg (comparable to a human dose) was injected intramuscularly in 6 chimpanzees, or when the PPF was heated at 60 degrees C for h and injected intravenously in 2 chimpanzees, there was no clinical or laboratory evidence of hepatitis B infection after 12 months, although 1 chimp of 2 who received the same material showed a borderline positive anti-HBsAg antibody result on one of 52 weekly serum samples. Since the new PE fractionation method is essentially nondenaturing, and simpler than the classical ethanol procedures, it was important to establish the noninfectivity of the final products.

Fusarium in wheat: importance and toxin production in the field - possibilities and limits of fungicide treatments.

Effects of plant cropping measures on toxin production were investigated. The use of the plough reduced the DON level by 71%. Additionally, the choice of less sensitive varieties, as well as an adequate nitrogen fertilization showed a significant reduction of the DON content in grain. In non-ploughed systems, an ear application with fungicides reduced DON by 63 %. The effects of strobilurin containing fungicides (strobilurin solo products and strobilurin-azole mixtures) upon the toxin content were compared with azoles.

Preparation of the major plasma fractions by solid-phase polyelectrolytes.

The solid-phase ethylene/maleic anhydride PE's E-5 and E-100 were used to developed a flexible integrated system of batch methods for the fractionation of albumin and gamma globulin as well as the clinically important coagulation factors. Each method for the isolation and purification of the fractions or a modification of it could also be used independently or could be added onto another method or methods. Each fraction was prepared in three steps: adsorption on solid-phase PE's elution from the PE's, and concentration by ultrafiltration or by use of PEG or both. Since the methods required no organic solvent but relied primarily on electrostatic bonding, they were essentially nondenaturing. Solid-liquid phase separation was easily accomplished by centrifugation or filtration. The yield and purity of each fraction was as follows: (1) coagulation factors II, VII, IX, and X concentrates 63% and 200-fold purified; (2) AHF 43% and 52-fold; (3) vWF 43% and 71-fold; (4) PPF, as albumin, 93% yield and 92% pure; (5) albumin 92% yield and 98.5% pure; and (6) gamma globulin 92% yield and 97.5 pure. During the preparation of gamma globulin the E-100 also tended to adsorb and remove the HBSAg and infectivity, when present.

Removal of hepatitis B surface antigen (HBsAg) from plasma fractions.

Endogenous or deliberately added hepatitis B antigen was removed and concentrated for assay from albumin, and from coagulation factor II, VII, IX, and X concentrates as model plasma fractions. The concentrates carry considerable risk of causing hepatitis in transfused patients. The amount of antigen remaining in the fraction was estimated to be less than 1/10,000 of that detectable by the Ausria II radioimmunoassay and 1/100 of that found to be infectious when highly contaminated human sera were diluted and injected in chimpanzees. Batch fractionation methods with polyethylene glycol were used. The yield of albumin was 96 per cent and of the coagulation factors about 90 per cent.