Therapeutic possibilities in Pseudomonas infections. Acute pseudomonas infections require treatment with antibiotics producing a bactericidal effect. The most useful are gentamicin, tobramycin, sisomicin and polymyxin B. In resistant strains, amikacin is indicated in addition. Carbenicillin, ticarcillin, carfenicillin or azocillin should never be given alone but in combination with some of the above preparations. Other drugs, such as chloramphenicol, tetracycline or streptomycin, though effective in vitro, should be avoided. Chemotherapy may be complemented by passive immunization either with hyperimmune specific gama globulin or hyperimmune plasma. A programmatic item of combined treatment is active immunization, especially with toxoid vaccine. Chronic processes are not, perhaps with the exception of urinary infections, suitable for antibiotic therapy. For this reason effective polyvalent vaccines should be developed from appropriate strains. It is now certain that in infections caused by mucous strains (most frequently encountered in cystic fibrosis) the vaccine should be prepared from these strains, since they have distinct functional and antigenic characteristics.


Investigation of the incidence of influenza A viraemia caused by virus strains circulating among children in 1968 - 1977. Nineteen strains of Type A influenza virus isolated from the blood of small children in 1968--77 were studied. The investigation of the strains in HAIR with antisera to the antigenic components of the strains in HAIR with antisera to the antigenic components of the strains  A/Hong-Kong/68,A/Anglia/72, A/Port Chalmers/73 and A/Victoria/75 made it possible to demonstrate antigenic "drive" of the haemagglutinin in the years 1968--1977 and to divide the strains into 4 varieties. A high sensitivity to inhibitors was observed in all the strains isolated. The study of pathogenicity and toxicity of the strains revealed viraemia in the strains isolated during the 1972--1973 epidemic and the subsequent epidemics with the absence of pathogenicity and toxicity for white mice. Regular finding of viraemia coincided in time with increased thermostablty of the haemagglutnin in the strains under study.


Testing the properties of an experimental batch of zoster immune gammaglobulin (ZIG). The methods of indirct haemagglutination (IH) and precipitation in gel (ID) were employed to test the level of varicella-zoster (VZ) antibodies in an experimental batch of zoster gammaglobulin (ZIG). The titre of indirect haemagglutinating antibodies in ZIG was about 64 times higher than in the ordinary batches of normal immunoglobulin and about 8 times higher in comparison with the level of the initial plasma pool. In the reaction of precipitation in gel, ZIG produced 5 to 6 zones. In comparison with the initial pool of convalescent plasma, ZIG also showed an 8-fold concentration of precipitating antibodies. ZIG was administered preventively to 6 children with risk diagnoses. None of the children fell ill with varicella. According to the results of subsequent serological examination in the reactions of indirect haemagglutination and radioimmunologic analysis, only 3 children were definitely susceptible to VZ infection. In two other children (very low antibody titres) the risk could not be excluded. No substantial increase in the levels of IH and RIA antibodies was observed in the 4 children under serological observation in a period of 6 months following the administration of ZIG. ZIG was administered therapeutically to four children with varicella. The effect of ZIG therapy was very suggestive, especially in two newborn infants lacking maternal antibodies, where the dose of ZIG per 1 kg body weigt was unusually high.


Experimental infection with Ancylostoma caninum larvae in mice. VI. Serum protein pattern use due to repeated exposures to infection. Three mice groups were infected with various repeated doses and other four groups with single doses of Ancylostoma caninum larvae. Serum samples collected at various intervals were analysed electrophoretically. The results were compared first with an uninfected control group and then with those of the corresponding non-immunized single-dose-infected groups. Remarkably decreased values of albumin, increased beta globulin and increased total globulins with a decreased A/G ratio were observed. There was no correlation between the decrease in albumin and the increase in beta globulin found in the immunized groups. No further enhancement in serum protein components was noted on repeated exposure to infection. Immunized groups of mice, infected twice with a 500-larval dose, showed an exceptional increase in alpha-2 globulin and unchanged gamma globulin levels.


The quantitation of cell surface antigens by antibody-complement mediated cytotoxicity: application to murine leukemia virus-infected mouse cells. 51Cr-antibody-complement mediated cytotoxicity (ACC) can be used to quantify nanogram amounts of cell surface antigens. When pure antigen is available for calibration, inhibition of ACC by known amounts of antigen and by whole cells yields an estimate of the number of antigenic equivalents per cell. ACC is more suitable for this purpose than radioimmunoprecipitation assay. By ACC the number of antigenic equivalents of the oncornaviral proteins gp70 and p30 has been determined on the surface of various cells infected with Rauscher and Gross murine leukemia viruses.


Purification of alpha-fetoprotein from mouse amniotic fluid by gel-entrapped antibody filtration. A one-step immunospecific affinity chromatographic method for purification of mouse alpha-fetoprotein (AFP) directly from amniotic fluid is described. The procedure employed a rabbit anti-AFP immunoglobulin-G entrapped in a polyacrylamide gel matrix prepared as a gel slurry. The presence of impurities could not be demonstrated in the eluted fraction and the recovery of AFP was approximately 26%. Sufficient quantities of AFP were readily purified for raising antibodies and for purity studies.


Phylogeny of the rabbit gamma-chain determinants: a d12-like antigenic determinant in Pronolagus rupestris. A survey for constant region gamma-chain allotypes (de locus) was undertaken in different lagomorph genera. As yet only Pronolagus rupestris, a paleolaginae, showed the presence of a determinant similar to rabbit d12 although it lacked the widespread e15 determinant. All seven individuals possessed the d12 like determinant which was studied by immunodiffusion, haemagglutination inhibition, radiobinding and binding inhibition assays. In addition, a new enzymic method for typing for d12, based on the presence of the asymmetric rabbit hinge carbohydrate linked to the d12 characteristic threonine, is presented. This method suggests, however, that, unlike the d12 rabbit, Pronolagus does not seem to have a majority of IgG molecules with a glycosylated hinge.


Biosynthesis of human blood group T-, N- and M-specific immunodeterminants on human erythrocyte antigens. We synthesized on Tn erythrocytes with human sera, UDP-Gal, and activators T-specific haptenic structures in satisfactory yield. The specificity of this biosynthesis was ascertained by agglutination with human and animal anti-T, by specific absorption of human anti-T as well as by agglutination inhibition assays. With isolated human erythrocyte T antigen as substrate we synthesized N- and M-specific structures with sera from individual human donors in presence of CMP-sialic acid by incubation for 24 hr at 37 degrees C. Serology on the recovered product was carried out with nineteen monospecific human and animal sera under strictly standardized and controlled conditions with the mandatory tube assay. All M- as well as N-derived T antigens tested acquired N specificity with all transferase sera of all MN types. In contrast, M-activation of M- and N-drived T antigens tested acquired N specificity with all transferase sera of all MN types. In contrast, M-activation of M- and N-derived T antigens occurred only if the transferase donor had the M gene. The nine M transferase sera used all gave M-activation of MM- and NN-derived T antigens. None of twelve transferase sera from NN donors M-activated any T antigen. NN antigen was transformed to a M-specific one by all transferase sera from MM donors but by none from NN donors. We have not yet established the biochemical-genetic relation of M to N; N may be the immediate precursor of M or M may originate directly from T. The sialyltransferase responsible for M activation may be a N transferase 'modified' by the M gene product or an entirely different sialyltransferase.


Differences in the susceptibility of MHC and non-MHC mixed lymphocyte reactions to suppression by murine amnionic fluid and its components. The ability of mouse amniotic fluid (MAF), alpha-foetoprotein (AFP) and MAF depleted of AFP (MAF - AFP) to suppress primary one-way MLR's was investigated. It was found that MAF, AFP and MAF - AFP were all suppressive of MLR's specific for MHC, K, D or I + S determinants. Suppression was observed when either lymph node or spleen cells were used as the responder cells. Nylon wool column passage of these cells did not significantly affect the immunosuppressive action of these substances. In contrast, MLR's specific for non-MHC/M-locus determinants demonstrated either diminished suppression or augmentation of the response, compared with the MHC stimulated MLR's. Our results show a differential effect of whole MAF and its fractions on the proliferative responses induced by various allogeneic stimuli and suggest that suppression is not due to a non-specific effect on proliferation regardless of the stimulus or cell type involved.


Radioimmunological determination of serum 3 beta-hydroxy-5-cholenoic acid in normal subjects and patients with liver disease. A radioimmunoassay for the determination of 3 beta-hydroxy-5-cholenoic acid in human serum has been developed, using 3 beta-hydroxy-5-cholenoyl-thyroglobulin as immunogen and 3 beta-hydroxy-5-cholenoylglycyl-125I histamine as radioactive ligand. The association constant was 6.3 X 10(8) l/mol. Cross reactivity with other bile acids of human serum was not detectable, but was 5.6% with cholesterol. Serum sample preparation included extraction of 3 beta-hydroxy-5-cholenoic acid from serum, solvolysis of sulfates, hydrolysis of conjugates, and separation from cholesterol by thin-layer chromatography. Serum concentrations of 3 beta-hydroxy-5-cholenoic acid were 0.23 +/- SD 0.12 mumol/l and 0.21 +/- SD 0.09 mumol/l in healthy males and females, respectively. In patients with primary biliary cirrhosis the serum concentration of 3 beta-hydroxy-5-cholenoic acid and the quotient 3 beta-hydroxy-5-cholenoic acid over total 3 alpha-hydroxy-bile acids (measured enzymatically) were significantly higher (P less than 0.02) than in patients with chronic active hepatitis or alcoholic cirrhosis. Analysis of 17 sera with elevated concentration of 3 beta-hydroxy-5-cholenoic acid by radioimmunoassay and capillary gas-liquid chromatography showed a close correlation (r = 0.91, slope = 0.97) between the results of the two methods.


Immunological relationships among ovine glycoprotein hormones. An antiserum to partially purified ovine FSH (but essentially free of LH) bound 125I-labelled ovine LH or bovine TSH. The antibody was directed exclusively against determinants in the alpha subunit. In a radioimmunoassay, only the intact ovine and bovine hormones and their alpha subunits were reactive; the hormone specific beta subunits exhibited no cross-reaction. The antibody directed against the alpha subunit was highly dependent on conformation. Human LH, FSH, TSH or their alpha subunits did not cross-react in the radioimmunoassay. Structural modifications such as acylation of the amino groups, reduction and alkylation of the S-S-bridges, or performic acid oxidation of the intact ovine FSH, LH or their alpha subunits virtually eliminated immunological reactivity. Using the alpha subunit radioimmunoassay, the presence of a significant quantity of free intact alpha subunit in standard (NIH) preparations of TSH-B7, FSH-S10 and LH-S19 was demonstrated.


Studies on antibodies against feline leukaemia virus (FeLV) in cat sera and rabbit anti-FeLV sera: cross reaction and differences. The indirect immunoferritin technique (IFT) that enables us to distinguish clearly whether an antibody reacts with a virus particle or only with the cell membrane, was used to study 25 cat sera and one rabbit anti-feline leukaemia virus (FeLV) serum using FL-74 cells as target. (1) All sera contained antibodies against FeLV even though 11 of the cats were viraemic  at the same time; (2) from the effect of glutaraldehyde fixation of the FL-74 cells on the reaction with cat sera and the results of blocking experiments, it could be concluded that cat sera and rabbit anti-FeLV sera react partly with different antigenic specificities of FeLV, partly with the same antigens; and (3) the indirect membrane immunofluorescence test using FL-74 cells as target is not a good test to detect the presence of antibodies against feline oncornavirus-associated cell membrane antigen (FOCMA) because FL-74 cells produce a large quantity of FeLV and the fluorescence measured could be from antibodies against FeLV.


Bidirectional axonal transport of free glycine in identified neurons R3--R14 of Aplysia. The axonal transport of 3H-amino acids was studied in the axons of identified neurons R3--R14 in the parietovisceral ganglion (PVG) of the mollusc Aplysia. The PVG was incubated (3--24 hr) in media containing physiological concentrations of single 3H-amino acids while the isolated nerve was superfused with plain or chemically altered media. The nerve was then sliced into sequential segments for biochemical analyses or fixed for autoradiography. 3H-glucine was transported at 70 mm/day in 6X greater quantities than other amino acids which were transported at less than 40 mm/day. In the 3H-glycine experiments, greater than 80% of the label transported into the nerve remained as free glycine, comigrating with glycine in thin-layer chromatographs. In autoradiographs of sections 4 mm from the ganglion-nerve barrier, greater than 50% of the silver grains were over R3--R14 axons which occupy less than 10% of the nerve cross-sectional area. EM autoradiographs confirmed that grains were within R3--R14 and not in surrounding glia. The selective transport of glycine was inhibited by Hg2+, by vinblastine and Nocodazole, and by low Ca2+ media. Autoradiographs of vinblastine-treated nerves showed a drastic reduction in label over R3--R14 and other axons. Label was also transported retrogradely; this transport rate was similar to the orthograde rate, but 5--10 times less label moved retrogradely. Autoradiographs showed that the retrograde label was localized to R3--R14 axons. This report clearly demonstrates the rapid, selective, and bidirectional transport of a free amino acid and provides further evidence that glycine may be used as a neurochemical messenter by neurons R3--R14.


Intracellular filaments in human cancer cells: a histological study. The distribution of intracellular filamentous systems in human breast and colonic cancers has been demonstrated by means of the tannic acid-phosphomolybdic acid-milling dye staining technique. Plasma membrane-associated staining is prominent in breast carcinomas and is strongest in anaplastic tumours. Strong staining is also noted in the cells at the margins of the tumours where the malignant cells are invading the surrounding tissues. In colonic carcinomas, filaments are mainly restricted to the terminal web region of the cells but dedifferentiation is accompanied by the development of circumferential staining of the cell membrane. The results are discussed in relation to immunohistochemical and electron microscopic studies of contractile proteins in non-muscle cells.


Intra-axonal diffusion of [3H]acetylcholine and [3H]gamma-aminobutyric acid in a neurone of Aplysia. 1. [3H]acetylcholine (ACh) or [3H]gamma-aminobutyric acid (GABA) was injected into the cell body of a cholinergic neurone of Aplysia kurodai. 2. [3H]ACh moved down the axon at a maximum speed of 2.5 mm/sec at 20 degrees C. 3. 20 mM-colchicine suppressed this movement, but some passive movement of radioactivity was noted along the axon. 4. Profiles of the passive movement coincided with theoretically obtained diffusion profiles. 5. The diffusion coefficient of ACh in the axoplasm was estimated. It was 3 x 10(-6) cm2/sec at 5 degrees C, 4 X 10(-6) cm2/sec at 15 degrees C and 6.5 x 10(-6) cm2/sec at 30 degrees C. The Q10 was 1.35, the activation energy was about 5 kcal/degrees C. These diffusion coefficients were lower than that of free diffusion of ACh (8 x 10(-6) cm2/sec at 18 degrees C, Fatt, 1954), and assumed to be reasonable, if one considers frictional resistivity of axoplasm in the diffusion of ACh molecules. 6. [3H]GABA diffused similarly to ACh, and the diffusion coefficients agreed with the estimated value when the molecular size differences were taken into account. 7. Both ACh and GABA seemed to diffuse in soluble form or as single molecules in the axoplasm. 8. Intra-axonal diffusion is very effective for short distances.


Ultrastructure of red cells frozen with hydroxyethyl starch. The freeze fracture appearance of red cells frozen in the presence of varying concentrations of hydroxyethyl starch (HES) is described. A technique is used which allows examination of a small portion of cells from a larger unit. The frozen cells appear distorted probably as a result of osmotic dehydration but indicate no evidence of intracellular ice. The frozen mixture with HES has three phases--a particulate phase consisting of the concentrated HES (and other salts), a sculptured ice phase and the red cells. When the concentration of HES is increased, the particulate phase becomes more prominent and at 14% HES appears to surround nearly all cells. In cells frozen in saline alone and 4% HES, the cytoplasm in a majority of cells has numerous cavities and depressions. Since such units haemolyse badly when thawed, it is possible that these regions indicate structural damage. In contrast, those units frozen with 14% HES (in which nearly 85% of the cells survive freeze-thaw) possess cells which only infrequently have such regions in the cytoplasm.


A simple apparatus for staining and washing a batch of grids. An efficient device for the staining and washing of up to fourteen grids simultaneously is described. It is simply constructed from inexpensive laboratory materials and is easy to load and operate.


Seronegative systemic lupus erythematosus. The diagnosis of systemic lupus erythematosus (SLE) often depens on positive tests for antinuclear antibody, LE cells, and antibody to DNA. In fact, the presence of such serological markers is virtually required for the diagnosis of active disease. This report describes 5 well-studied patients with SLE who initially presented with negative serology despite active multisystemic disease. When followed from 10 months to 7 yr, negative serologic markers, including the ANA, LE cell test, and anti-DNA were seen to change from negative to positive, analogous to the seronegative rheumatoid patient who may become seropositive. In a few cases of lupus, therefore, negative serologic tests do not always exclude SLE, and serial testing at periodic intervals may be indicated.


Extracellular aggregates of immunoglobulin in synovial fluid from rheumatoid arthritis. Particulate extracellular aggregates of immunoglobulin can be demonstrated in many whole synovial fluids from patients with rheumatoid arthritis but not in other diseases. These occur only in seropositive patients and correlate with more active inflammation and lowered synovial fluid C3. Aggregates were associated with intracellular inclusions but were more abundant and always contained both IgG and IgM. No aggregates were seen in 50 control fluids. These aggregates are compatible with immune complexes and should be included in studies of immunologic events in rheumatoid fluids.


Influence of an oversulphated heparinoid upon hyaluronate metabolism of the human synovial cell in vivo. Glycosaminoglycans appear to affect proteoglycan metabolism of fibroblasts, human synovial cells, and chondrocytes under in vitro conditions. The influence of an oversulphated heparinoid (Arteparon) on hyaluronate metabolism of the human synovial lining cell was studied by in vivo experiments on human volunteers. A higher production of better polymerized hyaluronic acid was observed after repeated intraarticular injections of this heparinoid into the knees of 2 osteoarthritic patients. After a single injection a statistically significant rise of synovial fluid hyaluronate concentration was observed over 4 days. A higher molecular weight of this molecule was suspected in view of a significant increase of the anomalous viscosity index 2 and 4 days after intraarticular administration.


[Immunocellular response to fetal antigens in patients with hepatoma (author's transl)]. The immunocellular response to fetal antigens was studied in ten patients with hepatocarcinomas. Homogenized extracts of human fetal liver and purified human alpha-fetoprotein were used as antigen substances. The control group included 15 patients with cirrhosis of the liver. The level of circulating T lymphocytes (E-rosettes) was also registered. Patients with hepatocarcinoma showed a definite response to both antigens, determined by the degree of inhibition of leukocyte migration. The migration indices were as follows: x = 0.65 +/- 0.16 for homogenized fetal liver antigen, and x = 0.79 +/- 0.13 for alpha-fetoprotein antigen. These values were 0.93 +/- 0.13 and 0.95 +/- 0.15 respectively in the cirrhotic patients. The differences in the migration indices for the two groups were statistically significant with both antigens (p less than 0.0005 and p less than 0.005). The decrease of the number of T lymphocytes in patients with hepatomas was also significant (p less than 0.005). The determination with homogenized fetal antigen was more sensitive than with alpha-fetoprotein (p less than 0.01). A significant relationship between the severity of the tumor and the immunocellular response could also be seen (r = 0.84; p less than 0.001). Response tended to diminish as the tumor progressed. The disappearance of immunocellular response seemed to depend at least in part on the decreasing number of T lymphocytes, since there was a significant inverse correlation between the two parameters (r = -0.75; p less than 0.01).


Randomised trial of jejunoileal bypass versus medical treatment in morbid obesity. The Danish Obesity Project. In a randomised clinical trial to assess the value of intestinal bypass in the treatment of gross obesity 130 patients who underwent end-to-side jejunoileostomy (with either a 1/3 or a 3/1 ratio between jejunum and ileum left in continuity) were compared with 66 non-surgically treated patients. All patients in the study had gross, long-standing, treatment-resistant obesity with resultant somatic, psychic, or social problems; none were alcoholic or had liver disease or other conditions which made them poor surgical risks. Most subjects were observed for more than 3 years. Median weight loss within 24 months was 42.9 kg in the bypass group, compared with 5.9 kg in the control group. No deaths occurred among those who underwent surgery. Patients who underwent intestinal bypass also had a better improvement in quality of life and a higher degree of patient satisfaction. Complications of the operation were, however, common and occasionally severe.


Double-blind trial of antihypertensive effect of chlorothiazide in severe renal failure. A randomised double-blind crossover trial was done to assess the efficacy of chlorothiazide as an antihypertensive drug in patients with severe renal failure. There was a significant reduction in standing (mean drop 13/6 mm Hg) and supine (mean drop 13/5 mm Hg) blood-pressure, without postural hypotension. Chlorothiazide has a place in the management of hypertension in patients with severe renal failure and its antihypertensive effect is probably due to a change in peripheral vascular resistance and not to volume contraction.


Dihydropteridine reductase deficiency diagnosis by assays on peripheral blood-cells. Human peripheral lymphocytes, granulocytes, and platelets contain the enzyme dihydropteridine reductase. A simple procedure and assay for diagnosing dihydropteridine reductase on peripheral blood is described. In twelve controls the level of enzyme activity in a lymphocyte-platelet preparation was 29.5 +/- 4.8 (SD) nmol NADH oxidised/min/mg protein. Two heterozygotes for dihydropteridine reductase deficiency had activities of 6.4 and 10.3 nmol NADH oxidised/min/mg protein, respectively.


Detection of virus-associated antigen in serum and liver of patients with non-A non-B hepatitis. In a search for serological markers of non-A non-B(NANB) hepatitis, sera from repeatedly transfused and convalescent patients were assayed by immunodiffusion against sera from 12 patients with early acute NANB hepatitis. A new antigen/antibody system distinct from HBsAg was demonstrated in 8 cases. To assess the specificity of the test, serial sera from 17 patients with acute hepatitis of known aetiology (10 due to hepatitis-B virus, 4 to hepatitis-A virus, 3 to drugs) were tested twice a month, together with sera from 14 NANB patients obtained during a prospective post-transfusion study. NANB antigen (Ag) was detected in at least one sample from 12 of the 14 NANB patients (86%) but in none of the other groups. NANB Ag appeared after or just before elevation of transaminase levels and was cleared before they fell to normal. 4 of 5 patients who showed seroconversion to NANB antibody (Ab) had transient hepatitis. In contrast, the alanine adminotransferase value returned to normal in only 1 of the 5 with persistent NANB antigenaemia during 6 months' follow-up. NANB Ag was also demonstrated by immunodiffusion in liver extracts from patients with chronic NANB hepatitis with antigenaemia. Fluorescein-isothiocyanate-labelled gammaglobulins with strong NANB Ab activity revealed specific nuclear fluorescence in foci of hepatocytes on cryostat sectons of these livers but in none of 6 control human livers. The results suggest that the antigen and antibody are specifically linked to NANB hepatitis of long incubation period.


Parenteral acyclovir therapy for herpesvirus infections in man. Acyclovir is a new antiviral agent which is highly active against herpesviruses in vitro and in laboratory animals. Twenty-three cancer patients with cutaneous and/or systemic herpes zoster or herpes simplex infections were treated with parenteral acyclovir. All had received previous specific treatment for their malignant disease and ten had undergone bone-marrow transplantation. The drug seemed to arrest the progress of the infections and was most effective when given early. Although two patients showed transient increases in blood-urea, possibly the result of acyclovir, the drug was remarkably non-toxic in the doses used.


Neurosecretory bodies within the median eminence in association with pituitary tumours. During necropsy the pituitary stalk is usually deliberately divided and the median eminence is separated from the posterior pituitary. In a series of 133 necropsies the brain was removed in a manner that preserved the integrity of the neurohypophysis and allowed histological examination of the entire neurohypophysis and the juxtaposed adenohypophysis. Neurosecretory bodies were found within the neurohypophysis in 2 patients,and in both microadenomas were found within the adenohypophysis. 2 other patients had larger tumours within the adenohypophysis but had no abnormalities within the neurohypophysis.


Dietary fibre, diabetes, and hyperlipidaemia. Progress and prospects. When diabetes and cardiovascular disease were first classed as possible fibre-deficiency diseases, laboratory and clinical evidence was lacking. Subsequent studies indicated that the gums and viscous types of fibre (e.g., guar and pectin) are more effective than other fibres in slowing carbohydrate absorption and hence in reducing the postprandial rise in blood glucose and serum insulin. This effect has longer term metabolic consequences. In addition, gums and viscous fibres reduce serum cholesterol, possibly by mechanisms other than simply increasing bile-salt loss. If these potential therapeutic effects of fibre are to be exploited, palatable formulations must be developed. The effect of fibre in whole foods should also be determined.


[alpha 1-Antitrypsin deficiency in early childhood (author's transl)]. alpha 1-Antitrypsin inhibits various proteases and excessive proteolysis. If serum and tissue concentrations of this compound are low throughout longer periods of time due to deficient synthesis, - which is a dominantly inherited trait, - progressive pulmonary emphysema will develop in adults, and liver disease in more than 50% of the cases in infancy and early childhood. Three subtypes can be distinguished: the heaptitis type, the intrahepatic hypoplasia of the bile duct system, and the cholangitic type which may imitate atresia of bile ducts. Prognosis of the liver disease depends upon the time, at which inflammatory processes stop, excessive proteolysis coming to a stop at the same time. No treatment is known for this disease.


[Portal hypertension - surgery during childhood (author's transl)]. Portal hypertension in children and infants is rare. It is caused in 75% of the cases by prehepatic obstruction of the portal vein, liver parenchyma being essentially unimpaired. Thus most of these cases may be cured, if the pathological hemodynamic situation can be corrected. The nature and extent of pathologic changes vary widely, thus the surgical approach has to be rather individual. The most important aim is to avoid esophageal bleeding but more has to be done to cure the underlying disease. 50 different surgical approaches have been described; when selecting one of these it has to be taken into account if and to what extent portal pressure has to be lowered, and how portal venous and hepatic arterial blood flow are to be influenced.


[Methods to prepare histological microautoradiographs for evaluation by automatic image analysis (author's transl)]. A prerequisite for automatic image analysis of histoautoradiographs is the arrangement of the silver grains within the same plane of focus. This can be obtained by semi thin sections of 1 micrometer in thickness. A sufficient contrast for the determination of the nuclear areas and the overlying silver grains at the same time can be reached by staining the preparations with Kernechtrot or Feulgen nucleal reaction. If a higher image contrast is desired sections measuring 2 micrometer or more in thickness may be subjected to the autoradiographic procedure without removing the resin.


[Selective and high-contrast staining in nervous tissue (author's transl)]. Using silver as an impregnating medium and controlling the staining process by physical development, a specific and reliable staining of high contrast can be achieved. These properties represent important prerequisites for automatic and quantitative television image analysis. Specificity depends on appropriate pretreatment. Television image analysis cannot only be used for object analysis, but also to quantify reliability and dependency of contrast on developmental conditions, as demonstrated for stained myelin.


[Portal hypertension associated with alcoholic liver damage (author's transl)]. Alcoholic liver damage is associated with the development of portal hypertension quite early, as a result of increasing fibrosis. By means of laparoscopic transhepatic manometry (LTM) in the branches of the portal and hepatic veins, we measured the pressure levels in 15 patients with early and transitional-stage alcoholic injury extending to cirrhosis of the liver, and compared them with histological and laboratory investigation criteria. We discovered that parenchymal damage with portal and centrolobular fibrosis already gave rise to some portal hypertension which, compared to a group showing histological changes of remodelling or cirrhosis, constantly increased. In completely developed alcoholic cirrhosis (n = 41), the pressure levels reached a peak. Despite this fact, bleeding from oesophageal varices cannot be predicted. Over the period of observation of 33 months, 4 deaths occurred (portal vein pressure between 4.5 to 5 kPa = 34.0 to 38.8 mm Hg), and three variceal bleedings we were able to manage were seen (pressures between 3.6 to 4.3 kPa = 27.0 and 32.0 mm Hg). Compared to hepatitic cirrhosis the prognosis was slightly more favourable.


[Diffusion of cefuroxime into the prostatic and other tissues of the urogenital region (author's transl)]. From 33 patients with prostatic adenoma with an average age of 68.4 years, serum and prostatic tissue concentrations after a single bolus-dose of 1.5 grams of cefuroxime were estimated. The mean serum concentrations 30 to 35 minutes after the injection were measured with 132.6 and after 55 to 60 minutes with 99.6 micrograms/ml. The corresponding concentrations in the prostata tissue were 29.2 after 30 to 35 minutes and 20.1 micrograms/g after 55 to 60 minutes respectively. After 145 to 155 minutes the concentrations in the serum were still at 23 micrograms/ml and 9.2 micrograms/g in the prostatic tissue. In other organ tissues of the urogenital region (renal tissue, testicle, fat, muscle) the concentrations were different depending on the state of the tissue. The estimated concentrations of cefuroxime in serum and tissue are suited for the treatment of infections caused by cefuroxime sensitive microorganisms.


[Evolution of viral diseases (author's transl)]. Biological characteristics of viruses determine their distribution in men and animals as well as their means of surviving or persisting. Taking measles as an example, we show that we are dealing with a recent phenomenon in evolution--as opposed, for instance, to those infections due to arboviruses (eg yellow fever) common in tropical regions. Survival rates are lower for highly virulent viruses (myxomatosis). Antigenmodification (influenza) or latency (herpes virus infections) allow the virus repeatedly to escape the body's immune system. The distribution and clinical appearance of the infections are influenced by environmental and particularly iatrogenic impact. Complete extirpation of infections will not be possible, only their manifestations will undergo continuous change.


Relationship between alpha-fetoprotein positive cells and precancerous lesions in rat liver during 3'-methyl-4-dimethylaminoazobenzene carcinogenesis. Localization of characteristic precancerous lesions was compared with that of alpha-fetoprotein (alpha-FP) positive cells during hepatocarcinogenesis in Wistar rats fed 0.06% 3'-methyl-4-dimethylaminoazobenzene in basal diet. Proliferative activity of liver cells was simultaneously followed by deoxycytidylate deaminase. The first alpha-FP positive hepatocytes were found after 3 weeks of carcinogenesis in certain parts of the capsule 0.1--0.2 mm thick. In the seventh week of carcinogenesis their number increased and they were present in groups also subcapsularly. At this stage, hyperplastic islets began to be formed in the liver parenchyma. After 14 weeks of carcinogenesis, alpha-FP positive cell clusters and islets were also found in the deeper layers of liver parenchyma in regions with proliferation of transitory cells. Hyperplastic basophilic foci and islets occurred regularly, however, these were not identical with alpha-FP positive sites and their incidence was more frequent.


Value of tumor markers in the treatment of endodermal sinus tumors and choriocarcinomas in the pineal region. A case of embryonal carcinoma in the pineal region of a 17-year-old boy is presented. The tumor included elements of choriocarcinoma and endodermal sinus tumor, and the use of human chorionic gonadotropin and alpha-fetoprotein as tumor markers is discussed. The markers were demonstrated both within the tumor and in the cerebrospinal fluid (CSF) and blood. The patient was treated with a postoperative program of irradiation and cancer chemotherapy, and at follow-up examination 20 months after operation no signs of residual tumor were present. It is suggested that human chorionic gonadotropin and alpha-fetoprotein should be measured in the blood and CSF before the treatment of midline tumors.


Spinal subdural hematoma: case report and review of the literature. A case of lumbar spinal subdural hematoma in a patient who had been on anticoagulant therapy is reported. Thus far 19 cases of spinal subdural hematoma have been reported in the literature, the majority in patients with a bleeding diathesis and after a lumbar puncture. Our case is the third reported to be in association with anticoagulant therapy. The hematoma was lumbosacral, in contrast to the usual location in the dorsal-lumbar area. A possible mechanism for the production of spinal subdural hematoma after a lumbar puncture is discussed. An early decompressive laminectomy and evacuation of the hematoma is the recommended treatment to obtain the best possible recovery of neurological function. (Neurosurgery, 5: 614--616, 1979).


Brain metastasis: current status and recommended guidelines for management. An overview of brain metastasis with respect to the pathological, diagnostic, and therapeutic aspects is presented. Management is almost always palliative, with cure being a rare exception. Evaluation of various therapeutic modalities--radiation, chemotherapy, or surgery--has been confounded by a lack of controlled, randomized studies whereby the relative benefit of the respective modalities can be assessed objectively. Despite these limitations, some progress is being made in the identification of those patients for whom therapy is likely to be of benefit. Apart from the use of steroids to control cerebral edema, radiotherapy is currently the most commonly employed therapeutic modality for cerebral metastasis. It is the treatment of choice for multiple intracranial metastases and it affords temporary improvement in neurological symptoms in about 60% of patients. For solitary metastases, combined therapy--surgical excision followed by whole brain radiotherapy--has been shown to result in a better quality and longer duration of survival than either modality alone. Except for patients who are terminally ill, aggressive treatment seems warranted, inasmuch as therapeutic results have been improving steadily over the years. Neither chemotherapy nor immunotherapy has been shown to be of benefit in the management of cerebral metastasis. An exception is choriocarcinoma, which responds well to a combination of radiation therapy and chemotherapy. Although the prognosis for meningeal carcinomatosis is poor, improved survival may be achieved by a combination of chemotherapy and radiotherapy. These are recommended guidelines for surgical intervention, usually followed by radiotherapy: (a) In general, surgical excision is recommended only for patients with relatively superficial, solitary lesions. It is reasonable, however, to consider the excision of a metastatic lesion that is immediately life-threatening or incapacitating, even though one or more other metastatic brain lesions may be present. This may be extended to the removal of multiple metastatic brain tumors if they are surgically accessible. (b) The second consideration is whether the primary tumor can or has been treated or if the primary tumor will permit reasonably long survival. (c) There should not be metastases elsewhere in the body, although their presence should not categorically exclude the patient as a surgical candidate. (d) The patient's general condition should be satisfactory. (e) Operation is recommended if the diagnosis of the intracranial lesion is uncertain. (f) A shunt should be considered for treatment of hydrocephalus secondary to obstruction of the cerebrospinal fluid pathway by tumor or edema. (Neurosurgery, 5: 617--631, 1979).


The value of smears in the diagnosis of gonorrhoea in the female. Of 332 consecutive patients attending the female venereal disease clinic, Auckland Hospital, 74 patients had gonorrhoea confirmed by culture, but gram stained slides were positive in only 36 of these patients. Five other patients had positive gram stained smears but were culture negative. The study showed the value of gram stained smear in the rapid diagnosis of gonorrhoea in the female.


[Combination of adriamycin and cyclophosphamide (alone or with other substances) in the treatment of breast cancer]. This paper reviews clinical trials at the University of Arizona Cancer Center which were designed to improve the outcome in breast cancer by utilizing the combination of adriamycin and cyclophosphamide (A--C) alone or with the addition of other agents or modalities. Our initial trial in advanced breast cancer with A--C produced an overall objective response rate of 78% in 51 patients with advanced breast cancer without prior chemotherapy. The median duration of disease control was 12 months. Subsequent studies showed that the addition of either vincristine or the androgen, calusterone, effectively doubled the remission duration and prolonged survival. In our surgical adjuvant trial with 6 months of treatment with A--C there has been only a 9% relapse rate in stage II patients thus far, with a median follow-up of close to 2 years. A subset of stage II patients who received regional radiotherapy along with A--C have not yet shown added benefit compared to the use of A--C alone. Since 1975, stage I patients have been treated with an abbreviated treatment schedule (3 courses of A--C over 9 weeks). While there have not yet been relapses in this category, much longer periods of follow-up will be required. The use of A--C (plus other drugs) has clearly provided excellent palliation and improved survival in patients with advanced or recurrent breast cancer; in our opinion it should be used as initial cytotoxic chemotherapy. The brief intensive program of A--C as a surgical adjuvant also shows considerable promise for erradicating occult micrometastases in both pre- and postmenopausal women.


Conjunctival transplantation as an alternative to keratoplasty. Rehabilitation of the ocular surface is frequently desired following severe chemical and thermal injury. While keratoplasty, both lamellar and penetrating, has been the usual mode of therapy, chronic healing deficits and graft rejections as a result of heavy vascularization often lead to an unsatisfactory result. In unilateral injury, conjunctival transplantation from the opposite, uninvolved eye offers an alternative therapy which avoids both intraocular surgery and the introduction of immunologically foreign tissue. Seven of ten cases of unilateral chemical or thermal injury showed marked reduction in vascularization and scarring following conjunctival transplantation. The characteristic early vascularization of the peripheral conjunctival grafts showed marked regression one to four months postoperatively. At the same time, the central cornea, whose superficial vessels and scarring were removed by keratectomy, showed reepithelialization and smoothing, without attendant vascularization. Three cases were optical failures partially as a result of deep scarring that could not be adequately excised with keratectomy. Preoperative evaluation must be directed primarily toward evaluation of the corneal thickness and the extent of deep vascularization to select those patients most likely to be improved by conjunctival transplantation.


Management of complications associated with lathe-cut soft contact lenses. At the present time, nine lathe-cut lenses are FDA approved and commercially available in the United States. Complications of lathe-cut soft contact lenses cover a wide spectrum of problems. Five significant complications of lathe-cut lenses and their prevention and therapeutic management are discussed.


Complications associated with aphakic contact lenses. Complications associated with rigid contact lenses are more frequent in aphakia. The limited dexterity of older patients not only may prove an insurmountable barrier to the mastery of daily contact lens insertion and removal, but also may produce significant direct eye trauma, its seriousness determined by the integrity of the cataract wound. Since prolonged wear of aphakic contact lenses is the rule rather than the exception, the risk of irreversible corneal changes such as vascularization and erosions is greater. A poorly fitted aphakic contact lens that does not permit adequate oxygenation of all corneal areas makes such complications even more likely to occur. Decreased corneal sensitivity associated with cataract surgery permits continued wear of a poorly fitted lens, which does not allow the cornea to recover between wearing periods. Detection of keratoconjunctivitis sicca, more prevalent in the older age group, may prevent the associated complications of corneal erosion, iritis, and infection. Complications of aphakic contact lenses can be controlled by patient instruction, appropriate contact lens adjustments, and regularly scheduled follow-up examinations.


Complications associated with therapeutic soft contact lenses. The complications of therapeutic hydrophilic contact lens wear are systematically outlined. Hypoxia is the major cause of initial patient failure as well as the long-term problems of corneal edema, neovascularization, and susceptibility to infection. The common difficulties with lens breakage, lens deposits, poor lens retention, and other entities are discussed. Suggestions for avoiding and treating various complications are given, including updated indications and contraindications for fitting these lenses. Careful patient selection and meticulous follow-up are essential.


Brief note. Induction of experimental allergic encephalomyelitis in sheep with purified bovine basic protein and adjuvant. A large animal model for the induction of experimental allergic encephalomyelitis to study the immunological disease parameters through closed-circuit extracorporeal thoracic duct filtration was successfully produced in two different strains of sheep. This study demonstrates the advantages of using a well-defined antigen to obtain both rapid onset of disease and a higher percentage of animals succumbing to the illness.


Anomalous alpha 2-macroglobulin-protease complexes in cystic fibrosis: decreased uptake of the complexes by fibroblasts in culture. Immunochemical and functional properties of control and Cystic Fibrosis (CF) alpha 2-Macroglobulin (alpha 2M) are compared. Crossed immunoelectrophoresis and Ouchterlony double diffusion revealed no qualitative differences between the two alpha 2-M preparations. Trypsin-esterase activity assayed with BAPNA as a substrate, in the presence of an excess STI, gave similar ratios between total and active alpha 2M. These alpha 2M-trypsin complexes were equally stable under various experimental conditions and maintained a constant STI non-inhibited esterase activity. Normal and CF-alpha 2M-trypsin complexes were taken up by normal human fibroblasts to a similar extent during a four hour period. The only significant difference was observed when the uptake of alpha 2M from untreated sera was examined. The uptake of alpha 2M from CF sera was always lower than from pooled control sera despite large variation. Mixing of control and CF serum did not affect the normal uptake and other serum components were taken up to the normal extent. Intracellular degradation of CF alpha 2M had a half life of 2.0 to 2.8 hours, which compares well to the normal half life of 2.2 hours. More work needs to be done on the nature of the interaction between alpha 2M and proteases before a reasonable explanation for the molecular nature of the abnormal behavior can be sought.


Two new serotypes of Erysipelothrix rhusiopathiae. Among a group of 16 argentine strains of Erysipelothrix rhusiopathiae 2 new serotypes have been found. Typing was performed by means of the agar gel diffusion test. Extracts produced by autoclaving the organisms for 1 hour at 120 degrees C were used as antigen. Antisera against all known types were produced in rabbits. Extracts produced from the two strains in question (Baño 36 and Baño 107) did not react with any of the knwon type antisera. Antisera against the two strains did not react with extracts of any of the known type strains, but only with extract of their homologous strains. The two new types were designated Type 21 (Baño 36) and Type 22 (Baño 107).


[Thrombocytopenic purpura during heparinotherapy. Two cases (author's transl)]. In two patients treated by heparin for thrombosis, thrombocytopenia under 10 000/cumm occured accompanied with bleeding cud recurrent thrombosis. The demonstration in the patients' serum of a factor inducing platelet aggregation and serotonin release in the presence of heparin suggests that the thrombocytopenia could be of immuno-allergic origin. In both cases, the discontinuation of heparin resulted in clinical and biological improvement. The rare cases of heparin-induced thrombocytopenia reported in the literature are discussed.


[Epstein-Barr virus associated Burkitt's lymphoma in a French child (author's transl)]. Pathologic diagnosis of malignant lymphoma, Burkitt type, was made in a 12 year old child with an abdominal tumour. The association of this lymphoma with Epstein-Barr virus was done based on the detection of Epstein-Barr viral markers within the tumour cells, as well as on the patient's serology. To our knowledge, it is the first time that the association between Burkitt's lymphoma and Epstein-Barr virus, which is frequent in Africa, is reported in a Caucasian French.


Myxobacterial hemagglutinin: a development-specific lectin of Myxococcus xanthus. Fruiting body formation in the bacterium Myxococcus xanthus consists of a temporal sequence of cellular aggregation and sporulation. During the period of cellular aggregation, a major new development-specific protein that has lectin-like activity is synthesized. This protein, called myxobacterial hemagglutinin (MBHA), was able to agglutinate sheep or guinea pig erythrocytes but not horse, ox, chicken, or human erythrocytes. MBHA was undetectable in extracts of vegetative cells, cells starved in liquid buffer, or in glycerol-induced cells. However, cells starved on a fruiting medium produced large amounts of MBHA (about 5% of protein synthesis), starting at about 6-8 hr of development. The protein accumulated in the soluble fraction of cells, reaching a peak of 1-2% of total protein at about the time when aggregation was completed. At later times the amount of MBHA present in the soluble fraction declined although synthesis continued. The hemagglutinating activity of MBHA could not be inhibited with simple sugars or aminosugars but could be inhibited with fetuin, a fetal calf serum glycoprotein. The O-glycosidically linked trisaccharide glycopeptide of fetuin was shown to be inhibitory by itself. The penultimate galactose of this glycopeptide was directly implicated in the inhibitory activity, because the inhibition by asialofetuin was reduced to 1/60th by periodate oxidation and to 1/15th after beta-galactosidase treatment. MBHA is an abundant biochemical marker of development in M. xanthus. The fact that it is a lectin suggests that it may play a role in cell-cell recognition or agglutination.


Structural studies of of "active complex" of bleomycin: assignment of ligands to the ferrous ion in a ferrous-bleomycin-carbon monoxide complex. Proton NMR studies at 360 MHz establish the binary Fe(II)-bleomycin complex to be paramagnetic with a spectrum covering 70 ppm. Addition of carbon monoxide generates a stable, diamagnetic Fe(II)-bleomycin-CO complex that is a putative structural analog of the "active" Fe(II)-bleomycin-O2 complex. The following six groups have been determined to be coordinated to the Fe(II) ion from analysis of the highly resolved 1H NMR spectra of this complex: CO, the primary and secondary amine nitrogens of the beta-aminoalanine moiety, the carbamoyl moiety on the 3-position of mannose, the pyrimidine N-1, and the imidazole N-1. The Fe(II)-bleomycin-CO complex binds to DNA, as shown by fluorescence quenching experiments, but Fe(II)-bleomycin-CO does not mediate thymine release. These results necessitate a major revision in the current model for metal coordination to bleomycin.


Calcium-dependent increase in adenosine 3',5'-monophosphate and induction of the acrosome reaction in guinea pig spermatozoa. Experiments were designed to determine the interrelationship between cyclic AMP and Ca(2+) during the processes of sperm capacitation and the acrosome reaction. In minimal culture media containing pyruvate and lactate as substrates, guinea pig spermatozoa required a minimum of 1.0-1.5 hr to capacitate in the presence of 1.7 mM Ca(2+) and a minimum of 0.5-1.0 hr to capacitate in the absence of added Ca(2+). Sperm cyclic AMP concentrations were increased by as much as 30-fold within 0.5 min after addition of cells to various media containing Ca(2+), and the concentrations then remained increased for up to 4 hr. When the cells were added to several Ca(2+)-deficient media, however, cyclic AMP concentrations increased only about 3-fold within 0.5 min and then returned to basal concentrations within 2 min. D-600, a calcium transport antagonist, completely blocked the Ca(2+)-induced increase in sperm cyclic AMP concentrations. In contrast to capacitation, the acrosome reaction failed to occur in the absence of extracellular Ca(2+). After capacitation of spermatozoa in a Ca(2+)-free medium, addition of Ca(2+) caused an increase in sperm cyclic AMP concentrations within 1 min and a maximal number of spermatozoa showing an acrosome reaction within 10 min. The addition of 1-methyl-3-isobutylxanthine along with Ca(2+) had a synergistic effect on the increase in cyclic AMP. Neither 1-methyl-3-isobutylxanthine nor 8-Br cyclic AMP induced an acrosome reaction in capacitated spermatozoa in the absence of Ca(2+), but both significantly decreased the time required for maximal expression of the acrosome reaction in the presence of Ca(2+). These results suggest that the sperm acrosome reaction is associated with both a primary transport of Ca(2+) and a Ca(2+)-dependent increase in sperm cyclic AMP concentrations. Because a cyclic AMP analogue did not induce an acrosome reaction in the absence of added Ca(2+), the increase in sperm cyclic AMP concentrations induced by Ca(2+) probably reflects one of a number of Ca(2+)-dependent events associated with the acrosome reaction.


Estimation of membrane potentials of individual lymphocytes by flow cytometry. The membrane potentials of individual cells can be estimated by flow cytometric quantitation of the cells' uptake of the fluorescent lipophilic cationic dye 3,3'-dihexyloxacarbocyanine iodide. Human lymphocytes separated from peripheral blood on Hypaque-Ficoll gradients are uniformly depolarized by gramicidin and hyperpolarized by valinomycin. Concanavalin A and phytohemagglutinin depolarize only a fraction of the lymphocytes. The flow cytometric technique allows precise detection of heterogeneous membrane potential responses to stimuli such as lectins; it could also provide a basis for sorting cells that respond differently to a given stimulus.


Dissociation and exchange of the beta 2-microglobulin subunit of HLA-A and HLA-B antigens. Human histocompatibility antigens HLA-A, HLA-B, and HLA-C are a complex of two noncovalently associated subunits: a heavy chain glycoprotein (alpha) carrying the genetic polymorphism and an invariant light chain, beta 2-microglobulin (beta 2m). Upon incubation of papain-solubilized HLA with radiolabeled urinary beta 2m, the latter is incorporated into HLA, where it substitutes for the preexisting beta 2m that has dissociated from the complex. The association-dissociation equilibrium that governs this beta 2m exchange reaction was investigated and found to be characterized by a long lifetime of the complex (half-life of 80 min at 37 degrees C) and a relatively low Kd (4 nM). The beta 2m exchange was used as the basis of a radioimmunoassay for HLA antigens with radiolabeled beta 2m as a unique label for all HLA specificities. In a similar fashion, radiolabeled beta 2m can be incorporated into HLA at the cell surface. Although the process is slower and less extensive than in solution, it can be used as a means to tag cells with specific probes for HLA antigens.


Disassembly of viral membranes by complement independent of channel formation. We have compared the effects of the complement membrane attack complex (MAC), nystatin, and melittin on the envelope of murine leukemia viruses to determine if channel formation alone is sufficient to cause membranolysis. Nystatin is a channel former and mellitin is not, although both are hemolytic. Whereas MAC and melittin disintegrated the viral membrane, nystatin had no effect on morphology, integrity, and infectivity of the virus. Incorporation of the antibiotic into the viral membranes was demonstrated by measurements of the characteristic fluorescence of nystatin in membranes and the dose-dependent increase in viral density after uptake of the antibiotic. The density of nystatin was measured to be 1.26-1.27 g/cm3. Proof for the formation of functional nystatin channels was obtained by light scattering measurements. Exposure of  untreated virus to hypotonic conditions increased viral light scattering because of osmotic swelling but otherwise had no effect on the integrity of the virus. Nystatin channel formation abolished the light scattering change, showing that the antibiotic had impaired the viral permeability barrier. We interpret these results to indicate that virolysis by MAC is not caused by channel formation and, conversely, in the absence of colloid-osmotic effects, channel formation by itself is not sufficient to disassemble a viral membrane.


Structural characterization of the murine fourth component of complement and sex-limited protein and their precursors: evidence for two loci in the S region of the H-2 complex. The S region of the murine major histocompatibility complex controls the expression of two related, serum substance-positive proteins; one (C4) has functional complement activity, whereas the other, the sex-limited protein (Slp), is hemolytically nonfunctional. The structural relationships of these molecules to each other and to their putative intracellular precursors have been examined. Radiolabeled intracellular C4 and Slp precursors were isolated from lysates of cultured peritoneal cells. The C4 and Slp precursors and their processed subunits were purified by immunoprecipitation and sodium dodecyl sulfate/polyacrylamide gel electrophoresis. Antigenically distinct precursors for C4 and Slp were demonstrated by sequential immunoprecipitation experiments in which anti-Slp-reactive molecules were precleared by exhaustive immunoprecipitation and residual C4 molecules were precipitated by antibody to serum substance. Both molecules had apparent molecular weights of 185,000. Their molecular identities as precursors of the mature C4 and Slp proteins were established in pulse-chase studies and by comparisons of their tryptic peptide profiles with those of isolated subunits from the processed proteins. When isolated alpha- or beta-subunits from C4 and Slp proteins were compared by peptide mapping, it was possible to detect multiple distinct and multiple shared peptides. This evidence indicates that the C4 and Slp proteins derive from distinct precursor polypeptides and suggests that the primary structures of the C4 and Slp alpha- and beta-subunits are different. These results support the postulate that the S region contains two discrete structural loci that specify discrete C4 and Slp proteins.


Crossreactive mixed lymphocyte reaction determinants recognized by cloned alloreactive T cells. We have isolated clones of alloreactive T cells by soft agar cloning of primed responder cells contained in serially restimulated, mixed lymphocyte cultures. These clones recognize unique mixed lymphocyte reaction (MLR) stimulating determinants present on (C57B/6 X A/J)F1 (B6A) spleen cells. By assaying the reactivity patterns of these clones on a panel of stimulator cells, we have identified at least three separate "specificities" of alloreactive T cell clones. Several clones exhibiting one of these three reactivity patterns have been isolated on many occasions from different experiments. Using such clones of alloreactive T cells, we have identified and are attempting to define crossreactive MLR stimulating determinants exhibited by a panel of stimulator cells. Recognition by certain clones of a shared MLR determinant(s) on B6A and DBA/2 and of the lack of this determinant(s) on B6 stimulator cells questions current concepts concerning the nature of MLR stimulating determinants and T cell recognition of alloantigens.


Induction of calcium flux across the rat mast cell membrane by bridging IgE receptors. Rabbit antibody against IgE receptors or its F(ab')2 fragments induced an increase of 45Ca uptake into normal mast cells, and this process was accompanied by histamine release. Evidence was obtained that a substantial portion of 45Ca uptake was due to movement of calcium across the cell membrane. Neither 45Ca uptake nor histamine release was induced by Fab' monomer fragments of the antireceptor antibody. However, bridging of receptor-bound Fab' fragments by antirabbit IgG or bridging of receptor-bound IgE by anti-IgE induced an increase in 45Ca uptake. The results collectively indicate that crosslinking IgE receptors increases the membrane permeability of the mast cells toward calcium.


Antigen-induced strain-specific autoantiidiotypic antibodies modulate the immune response to dextran B 512. Sera from CBA and C57BL mice immunized against dextran B 512 eight or more days earlier contain mainly IgG antibodies that specifically suppress the development of plaque-forming cells against dextran in vitro. Suppression was not caused by antibodies against the alpha 1-6 epitope of dextran to any major extent. Sera from CBA mice (IgCH allotype j) suppressed plaque-forming cells from CBA mice, whereas sera from C57BL mice (allotype b) inhibited plaques from C57BL and CBA/Ig-1b mice. Sera from dextran-tolerant mice or from mice that are genetic unresponders to dextran did not suppress plaque-forming cells. Both thymus-independent and thymus-dependent forms of dextran induced the appearance of these autoantiidiotypic antibodies that can distinguish between anti-alpha 1-6 antibodies produced in mouse strains having different IgCH-locus-determined allotypes.


Effect of colicins Ia and E1 on ion permeability of liposomes. Colicins Ia and E1 are shown to inhibit the formation and bring about the collapse of a potassium diffusion potential imposed across the membrane of liposomes prepared from soybean or Escherichia coli phospholipids. Such depolarization results from a colicin-induced increase in membrane ion permeability. Colicins E2 and E3 do not depolarize such membranes. In addition to the colicin Ia-induced rapid efflux of preloaded rubidium, sodium, phosphate, or choline from liposomes, a slower efflux of preloaded sucrose or glucose 6-phosphate occurs. However, treated liposomes do not leak inulin or dextran, demonstrating that the effects of E1 and Ia are not due to a general disruption of membrane structure. The fact that colicin-induced ion efflux is observed in the complete absence of a membrane potential shows that the action of these colicins on liposomes is not voltage dependent. These results provide strong evidence that the depolarization of E. coli cells by colicins Ia and E1 results from a colicin-induced increase in membrane permeability to ions. It is proposed that this is brought about by the direct interaction of the colicin molecules with the bacterial cytoplasmic membrane.


Estrogenic activity of the insecticide chlordecone (Kepone) and interaction with uterine estrogen receptors. The chlorinated insecticide chlordecone (Kepone) interacts with the estrogen receptor system in the rat uterus in vitro and in vivo. It competes with estradiol for binding to the cytoplasmic receptor in vitro and also induces nuclear accumulation of estrogen receptor sites in uteri in vitro. When injected into immature rats, chlordecone translocates estrogen receptor sites to the uterine nucleus, increases uterine weight, and stimulates the synthesis of the progesterone receptor, an estrogen receptor-mediated process. Its slow onset of action but prolonged duration of interaction with estrogen receptor and stimulation of uterine weight gain and progesterone receptor synthesis indicates that, although it has an affinity for receptor only 0.01-0.04% that of estradiol, its considerable estrogenic activity may likely be derived from its long half-life and bioaccumulative character.


Immunotherapy in atopic disorders.

Immunotherapy in atopic disorders. Immunotherapy plays an important role in the treatment of allergic rhinitis, asthma, and insect sting allergy. In the latter circumstance, immunotherapy with specific venom from the stinging insects can be life saving. Most commonly employed in immunotherapy are aqueuos extracts of aeroallergens, particularly plant pollen, and mold spores. Recent advances suggest that modification of these allergens, such as polymerization, can result in improved symptom response with fewer side effects. Efficacy trials of immunotherapy have demonstrated satisfactory treatment responses in allergic rhinitis and allergen-induced asthma. These responses, as measured by changes in symptom scores, are attended by specific immunologic changes summarized in Table 2.


Analysis of factors which might improve the survival rate of infants with congenital heart disease. During 1973--1977, 212 infants underwent open heart operations for congenital heart disease at the Thoracic Unit, The Hospital for Sick Children, Great Ormond Street. The current results (75% survival rate) are compared with the results from earlier years (1963--1967) which showed only a 35% survival rate. Two groups are discussed in detail: Infants with Total Anomalous Pulmonary Venous Drainage, and Persistent Truncus Arteriosus. Although the operative risk in both groups remains high, the analysis of the natural history, palliative and corrective surgery and causes of death suggests that early total repair of severely symptomatic infants achieves the optimal results. The importance of careful analysis is emphasised. Palliative surgery still plays an important role in the treatment of congenital heart defects in infancy. The optimal timing and time of operation can only be selected after careful analysis of various factors.


Characterization of two pregnancy associated serum proteins with pregnancy specific beta 1-glycoprotein (PS-beta-G)-determinants. The existence of two pregnancy associated, partially identical glycoproteins with PS-beta-G determinants were demonstrated in serum from pregnant women. Crossed and crossed-line immunoelectrophoretic analysis of pregnancy sera revealed two precipitates with alpha- and beta-mobility, respectively. The ratio between the precipitation areas in various samples could be related to three different types of precipitation patterns observed in rocket immunoelectrophoresis. This variation in precipitation pattern could so far not be related to the progression of any obstetric parameters. The diphasic shape of the precipitate in crossed immunoelectrophoresis was not changed by incubation with normal human male serum or antibody to normal human serum. Incorporation of 4% PEG in the gels enhanced the precipitate in the alpha-region. The cross-reactive glycoproteins were detected in all pregnant women examined. These results point to the need for specific antibody preparations permitting separate quantitation of the two antigen populations with PS-beta-G determinants.


Serum beta2-microglobulin in liver disease. The concentration of beta 2-microglobulin in serum was determined in seventy-one patients with various liver disorders. Elevated values were found in most patients with chronic active or chronic persistent hepatitis and in over 80% of patients with alcohol-induced liver cirrhosis. In contrast, patients with alcohol-induced fatty liver, the serum beta 2-microglobulin concentrations were mostly within the normal range. Significant correlation (P less than 0.001) was noted between the elimination rate of galactose from blood and the serum beta 2-microglobulin concentration in patients with alcoholic liver damage but not in patients with chronic hepatitis. The reasons for the increased S-beta 2-microglobulin concentrations in liver diseases are unknown. Several explanations including a release of beta 2-microglobulin from necrotic liver cells or an increased synthesis of beta 2-microglobulin consequent to inflammation in the liver are possible. Alternatively, raised beta 2-microglobulin levels may reflect the hepatic synthesis during reparative growth.


[Short-term effects of gamma-globulins on sleep organization in epileptic patients (author's transl)]. A study of the effect of gamma-globulins on sleep was conducted in 8 patients aged 1 to 35 years with epilepsy not responding to any of the conventional treatments. Two polygraphic recordings were carried out during the 4th and 5th nights following a placebo injection and then an injection of gamma-globulin 16 Merieux (1 ml/kg). The short-term effects of the gamma-globulins were : --reduction in percentage of paroxysms, --an acceleration in the electroencephalographic tracings of the different stages of sleep and those of the waking period, --a statistically significant increase in the percentage of paradoxical sleep, though the general organization of sleep remained unchanged.


Hormonal influences on the dog prostate--a correlation study with light microscopy, transmission and scanning electron microscopy. Dog prostates under defined hormonal influences were investigated by light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The normal acinar cell apices exhibited well-developed, densely packed microvilli. TEM demonstrated numerous secretory granules, indicating a high secretory activity. Following castration, the acinar cells of the dog prostate showed rounded apices. Most of the microvilli disappeared and only knob-like protrusions were seen. Castrated dogs following administration of 3 alpha-androstandiol showed fewer microvilli than the controls; however, a well-developed undulating surface could be observed. Castrated dogs given 3 alpha-androstandiol and cyproterone acetate showed almost completely atrophic glandular cells. Prostates of dogs with spontaneous benign prostatic  hyperplasia (BPH) were characterized by pleomorphism of cell shapes and surfaces. The different hormonal influences induced changes of the prostatic surface structures and secretory activity.


Comparative scanning and transmission electron microscopy of the prostatic gland in different species (mouse, rat, dog, man). Man, dog, rat and mouse prostatic glandular cells were compared by light microscopy (LM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). In rodents and dogs, a merocrine secretion exists, and in these species the undulating surface represented a confluence of secretory granules. The prominent secretory blebs observed in dog and human benign prostatic hyperplasia (BPH) corresponded to apocrine secretions. Also variations in the amount of secretory activity were seen. Androgenic hormones were responsible for the differentiation of the prostatic epithelial cells during development. Therefore, an absence of microvilli as well as the appearance of ruffles, microplicae, and bare cells were seen in the prepuberal and castrated rat. In the elderly rat and in human BPH, a pleomorphism of the apical cell surface was evident.


Visualization of subsurface structures in cells and tissues by backscattered electron imaging. A fraction of the beam electrons which interact with a specimen scatter back. The number of backscattered electrons (BE's) increases with the atomic number of the elements encountered. Cell and tissue structures lacking a heavy metal content yield few BE's compared to structures affixed with heavy metals, either vitally or by means of staining methods applied after fixation. The BE imaging mode of a scanning electron microscope (SEM) provides an intensity map of the BE yield from the specimen. BE imaging of selectively stained structures in cells and tissues renders these structures visible in contrast to the unstained surround. Since BE's can emerge from a significant depth within the material, BE imaging can be used to view such heavy metal stained structures beneath intact cell surfaces. The microcontours of the overlying surface can be viewed concurrently by using the surface scanning (i.e., the secondary electron imaging; SEI) mode of the microscope. Methods for selectively contrasting subsurface structures can be adapted from existing light microscope (LM) and transmission electron microscope (TEM) methods. Staining methods have been devised for subsurface viewing of cell organelles, including nuclei, mitochondria, peroxisomes, lysosomes, and phagosomes. A physical model is presented which describes these observations and suggests future possible trends in this subject. Specifically the image contrast and resolution are described in terms of the physical properties of the stain and specimen and of the SEM operating conditions of energy and current. Finally a summary of instrumentation considerations describes present and potential BE detectors, their ancillary electronics, and image processing.


Cross-reactivity of cytotoxic T-lymphocyte antigen-binding receptors immune to antigens of the H-2 system. Anti-B10 D2 (anti-KbDb) cytotoxic T lymphocytes (CTL) and monospecific anti-Kk and anti-Dd CTL, separated from their mixture in anti-A B10 lymphocytes and concentrated with the absorption-elution technique on the macrophage monolayers of various genotypes, gave rise to cross-lysis of third-party target cells. Anti-B6 R101 (anti-Kb) CTL, which display a cross-lysis of mutant Kbd (M505) and Kba (Hzl) target cells, were separated into two fractions, one of them being able and the other unable to adhere to the cell monolayer of the particular mutant. The fraction of anti-Kb CTL adherent to Kba antigen was isolated by elution from the Hzl cell monolayer. These anti-Kba CTL and CTL eluted from the B6 cell monolayer displayed a considerably higher cytotoxic effect on B6 than on Hzl target cells. This difference was not due to lower sensitivity of Hzl targets to lysis. The findings indicate that anti-Kb CTL are homogeneous in the specificity of their receptors. The H-2 molecule is presumed to bear a single CTL determinant only, which induces a CTL clone or a group of clones homogeneous in specificity but heterogeneous in the affinity of its receptors.


HLA-D-restricted antigen activation of sensitized T lymphocytes: studies on the ability of HLA-D/DR-expressing B lymphocytes to substitute for macrophages in antigen activation. The proliferative response of sensitized human T lymphocytes to purified protein derivative (PPD) and to trinitrophenyl (TNP)-conjugated autologous cells in vitro is restricted by self HLA-D/DR determinants. Here we report that the PPD-specific response is strictly related to the content of phagocytosing cells (macrophages, Mphi) in the cultures and that an optimal PPD response occurred at a T/Mphi ratio between 10:1 and 5:1. B-cell-enriched suspensions, which also express the HLA-D/DR determinants, were not able to replace the macrophages in this HLA-D/DR-restricted response. On the other hand, TNP-treated similarly prepared B cells were in most instances effective in inducing a secondary TNP-specific response of in vitro-sensitized T cells.


Pneumococcal infections and the possible need for a vaccine. Antibiotics and chemotherapeutics have been available for about 40 years, but still bacterial infections constitute some of the greatest problems in medicine. Pneumococci causing pneumonia, bacteremia, meningitis and otitis are a leading cause of illness and death. The exact incidence and lethality of pneumococcal infections is not known, however, since they are not reportable diseases in most countries and since microbiological diagnosis is difficult. In the latest years some significant progresses have been made for the diagnosis of infections caused by pneumococci, especially pneumonia. This is for example the counterimmunoelectrophoresis (CIE) for antigen determination, the transtracheal aspiration (TTA) for direct bacterial cultivation from trachea, and serological assays like radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) for pneumococcal capsular antibody determination. These techniques have further emphasized the significance of pneumococcal infections. The recent finding of pneumococci resistant to penicillin and some other antibiotics also emphasizes the need for immunological prophylaxis. In recent years a vaccine consisting of the purified, most common pneumococcal polysaccharides has been introduced. It has been shown to protect against pneumonia, pneumococcal infections in splenectomized individuals and people with spherocytosis and probably partly against otitis media. Indications for the vaccine are suggested.


In vitro aggregation of mouse peritoneal exudate cells. (1) Immunological mechanisms which cause macrophage agglutination in vitro have been investigated. (2) Quantitative differences in the agglutinating properties of sero-positive rheumatoid sera and normal sera were found. (3) NZB X W mouse peritoneal macrophages differed from normal (NZCW) mouse peritoneal macrophages in susceptibility to agglutination. These quantitative differences could be explained by NZB X W macrophages carrying a surface immunoglobulin component which increased agglutination by rheumatoid sera. (4) Antimacrophage antibody was more effective in rendering macrophages susceptible to the agglutinating effect of antiglobulins than was heat-aggregated gamma globulin.


Some membrane events occurring during fusion and exocytosis in rat peritoneal mast cells. We have used the mast cell as a model system for studying some of the membrane events which occur during exocytosis. Our observations indicate that the maximum cluster size of IgE molecules necessary for the 'on' signal to activate a mast cell is ten or less, and that the 'off' signal is not associated with the gross patching or pinocytosis of IgE and its Fc receptors. Furthermore, the use of Con A-Sepharose beads to stimulate mast cells has shown that such signalling is localized to the areas of stimulus, but this localization is not a function of desensitization over the rest of the cell since the subsequent addition of soluble Con A to locally released cells induced generalized degranulation. Ca2+ influx therefore acts in a localized manner to initiate degranulation. Following receptor cross-linking, most of the membrane proteins and the layer of intervening cytoplasm are laterally displaced away from the areas of membrane interaction. This displacement may act as the signal for fusion to occur. The resulting fused bilayers are predominantly lipid, a situation which may be common in all transient membrane fusion.


H-2L: demonstration of four new allelic products and independence of H-2D and H-2L molecules. The H-2L molecules were detected for the first time in the Dd region products using antisera against the public specificity H-2.28. This specificity was analyzed because its presence in K as well as in D region products and its apparent allelism with another public specificity, H-2.1, indicated that these two specificities may have a special position may have a special position in the H-2 system. This was corroborated by subsequent identification of H-2L molecules in Dq and Dk products using anti-H-2.28 AND ANTI-H-2.1 sera, respectively, while none of the other previously known public or private specificities was detected on H-2L molecules. We tested the products of four D region alleles which had not been analyzed previously. In each of them we identified two distinct types of molecules: H-2D, which reacts with sera agains the D region private specificity, and H-2L, which does not react with these sera, but which is detectable either by anti-H-2.28 sera (H-2Lb, H-2Lf, H-2LS) or by anti-H-2.1 sera (H-2Ldx). This increases the number of identified H-2L alleles to seven (five H-2.8+, two H-2.1+). No association between H-2D and H-2D and H-2L molecules on the cell surface was detected in capping experiments.


HLA markers in six Lebanese religious subpopulations. A total of 315 normal, unrelated individuals from six of the seven principal religious communities of Lebanon have been typed for 26 alleles of the A and B loci. The gene, haplotype frequencies and delta values have been calculated. The HLA gene frequencies for loci A and B reported in this study are shown to cross Moslim-Christian lines, but generate a distinguishably different profile for the Armenian immigrant subpopulation. This first-level homogeneity which is inconsistent with results earlier reported for ABO, rhesus and Gm systems, seems to disappear when linkage disequilibrium deltas are compared.


Some structural properties of thymus leukemia antigen (TL) solubilized with detergent. Plasma membranes were isolated from the leukemia cell line ASL1w and extracted with detergent (DOC). DOC solubilized more TL activity than could be detected on isolated membranes. However, extraction of membranes with LDS or EDTA solubilized only 17% and 4%, respectively, of the activity. This indicated that TL was not loosely associated with the membrane but rather was integrated into the lipid bilayer. At low concentrations of DOC (0.05%), TL was found to be largely aggregated and was also prone to autolysis. Neither aggregation nor autolysis was observed at a higher DOC concentration (0.5%). The apparent molecular weight of TL in 0.5% DOC was determined by Sephadex G-200 chromatography to be about 65,000-70,000. Digestion of a 0.5% DOC extract of TL with either papain or trypsin produced a fragment of TL of about 35,000 molecular weight. These fragments were similar in size to a fragment produced by autolysis. These data suggested that a region of the TL molecule was very prone to proteolytic attack. The 35,000 molecular weight proteolytic fragments bound specifically to lentil lectin affinity columns, which indicated that they retained at least part of the carbohydrate present on the native molecule.


A method for the differentiation of T and B lymphocytes and monocytes migrating under agarose. This report describes alterations in the agarose lymphocyte migration technique which resulted in satisfactory differentiation of T and B lymphocytes and monocytes which have migrated as a monolayer for 1-3 days. The Wright-Giemsa staining used in the original method did not permit identification of individual migrating cell types. The most important modifications were changing from a plastic to a glass migration surface, and significantly reducing the overlying thickness of agarose which permitted a short fixation time and easy preparation of permanent slides stained for nonspecific esterase. The esterase staining of monocyte cytoplasm was intense and diffuse. One or two small, discrete areas of cytoplasmic esterase activity were identified in the majority of T lymphocytes. B lymphocytes showed either a trace or no evidence of esterase activity. The modified method should prove useful for the histochemical differentiation of migrating subpopulations of mononuclear cells.


Rapid removal of acid from glycol methacrylate for improved histological embedding. A method for the rapid and complete removal of methacrylic acid from commercial samples of glycol methacrylate is presented. It entails conversion of the acid to an insoluble N-acylurea by treatment with an equivalent amount of N,N'-dicyclohexylcarbodiimide. Sections of tissue embedded in polymer prepared from the purified monomer can be stained with basic dyes without simultaneously staining the polymer.


A method for myoglobin in cryostat sections of muscle by precipitation with sulfosalicylic acid. Transverse cryostat sections of skeletal muscle were fixed in a solution containing 1.5% glutaraldehyde and 1.5% sulfosalicylic acid and stained in a solution containing equal volumes of 3% hydrogen peroxide and 50% ethanol saturated with o-tolidine. Myoglobin in the sarcoplasm of muscle fibers was precipitated and stained blue. Applicability of this method to cryostat sections, without glutaraldehyde fixations prior to freezing, allowed the myoglobin content of individual muscle fibers to be correlated with other histochemical characteristics of the same fibers seen in serial sections. In the dark red bovine sternomandibularis muscle, fibers with weak adenosine triphosphatase (ATPase) and strong succinate dehydrogenase (SDH) activity always exhibited strong myoglobin staining. An equal degree of staining was found in many fibers with strong ATPase and intermediate to strong SDH activity. Fibers with strong ATPase and weak SDH activity were less strongly stained than the preceding types.


Contributions to semithin sectioning on a conventional rotary microtome. Procedures for obtaining sections 1 micrometer thick on a conventional rotary microtome are described. Hydrophilic resin blocks with adequate hardness and elasticity for semithin sectioning are made by addition of divinylbenzene and methylmethacrylate to a commercial embedding kit. The blocks are pinched between two simple adapters and mounted in the specimen holder of a microtome. A glass knife of the Ralph type with an effective blade length of 25 mm is made from a glass slide and attached to a metal bar with paraffin. The low cost assembly is set in the steel knife holder of a conventional rotary microtome. Sections 1 micron in thickness can be cut from the resin embedded blocks. Staining with the usual staining solutions may be weak due to the thinness of the sections, but the fine resolution and low distortion achieved are compensating gains.


Trypsin-orcein banding in plant chromosomes. A convenient and quick method using trypsin-orcein for banding plant chromosomes (O-banding) is suggested. The technique is directly applicable to meristematic tissues (e.g. root tips) and involves the treatment of root tips with 1-2% solution of trypsin either in buffer or in 0.5 N HCl for 5-10 minutes at 37 C or for 30-60 minutes near 0 C followed by staining with 1.5% acetic orcein: 1 N HCl (19:1). Dark staining bands are reproducible and species specific. These bands possibly represent specific DNA-protein-dye interaction.


Nucleoli visualized by silver staining combined with a new RNA-specific fluorochrome. A technique for distinguishing nucleolar components by light microscopy is described. Silver staining is used for localizing nuceolar-organizing regions. A new RNA-specific fluorochrome, 4''-6-bis(2'-imidazolinyl-4'-5'H)-2-phenyl-4'-phenoxyindole, visualizes the RNA-containing "partes fibrillares et granulares" of the nucleolus. Both staining procedures can be combined to analyze nucleolar structure and function by light microscopy.


A rapid technique for preparing microorganisms for transmission electron microscopy. A rapid and efficient method of preparing microorganisms for transmission electron microscopy is reported. In developing the method Salmonella, streptococcal, and protozoal specimens were fixed with glutaraldehyde. After fixation cells are collected on a membrane filter, washed with buffer, postfixed with osmium tetroxide, then washed with distilled water and stained en bloc with uranyl acetate. Specimens are dehydrated using a graded series of acetone and then infiltrated with graded mixtures of acetone and Spurr embedding medium. Finally the membrane filter is cut into small pieces and embedded in fresh embedding medium polymerized in polyethylene capsules. By collecting and processing the specimens on membrane filters, numerous centrifugations are eliminated from standard procedures. The use of a low viscosity embedding medium allows for rapid infiltration and embedding of the specimen. Using this technique microbial specimens can be sectioned after less than 4 hours preparation.


[Individual variability and an error in the method of determining protein thermostability]. The individual variability of the heat resistance of two enzymes (aldolase and actomyosine) of the m. gastrochemius of the grass frog Rana temporaria has been quantitatively evaluated by means of analysis of variance. 37--41% of the total variability of the character may be due to errors in the methods of investigation, whereas 59--63% of the variability is due to phenotypic differences between individuals of the same population. The extent of error is primarily associated with the initial stages of the experiment, i. e. the isolation of the enzyme preparations. The individual variability of the heat resistance of proteins is lower than that of the cells or the organisms from which they are derived.


[Antitumor resistance induced by immunization with normal definitive tissues]. Immunization of C3HA mice with homogenates of various normal definitive tissues, obtained from syngeneic, allogeneic and xenogeneic donors, resulted in an increase of antitumor resistance, as evidenced by a considerable retardation of growth of the transplantable strain-specific hepatoma 22a. The effect may be due to sensibilization of animals to the antigens of normal definitive cells, and, in particular to the tissue-specific antigens characteristic of cells of the tumor in question.


Effect of hemodialysis on plasma beta thromboglobulin levels. BTG levels are significantly elevated in patients on chronic hemodialysis. The increased levels are not due to turbulent flow within the vascular access, but may be due to platelet aggregation within the hemodialyzer during the first hour of dialysis. This effect is observed with the Travenol CF series hollow fiber (cuprophane) but not the Cordis-dow artificial kidney (regenerated cellulose).


[Soluble Babesia ovis antigen].

[Soluble Babesia ovis antigen]. Dissolved antigen of plasma and of erythrocytic cytoplasma was obtained from 4 splenectomized sheep experimentally infected by Babesia ovis. Blood was taken in the stage of acute parasitemia with 23--25% parasite erythrocytes. The antigen was isolated by precipitating the plasma and the erythrocytic cytoplasma three times with an ammonium sulfate solution saturated 50 and 75%. The fraction obtained following dialysis was concentrated after McErlean's method. The specificity and activity of the isolated soluble antigen were proven by RAGP against serums of 6 weaned lamb groups: I group -- with 4 cm3 plasma, II group -- with 8 cm3 plasma, III group -- with 2 cm3 erythrocytic cytoplasma, IV group -- with live vaccine, V group -- with normal plasma and VI group -- normal weaned lambs. The same groups were used to assess the immunogenic properties of the dissolved antigen. Following provocation with a virulent B. ovis strain the lambs of group II proved as resistant as these immunized with live vaccine. The result was confirmed under production conditions -- not one of the 360 lambs immunized with 5 and 10 cm3 plasma became infected, while of the 250 control lambs 30 were infected. The conclusion is drawn that plasma in the stage of acute parasitemia can be used for immunoprophylaxis, but more supplementary studies are needed.


Glycosomes (protein-glycogen complex) in the canine heart. Ultrastructure, histochemistry and changes induced by acidic treatment. Cardiac conducting fibers were selected from two dogs defined as A and B. The specimens differed in the reaction of their electron dense granules, commonly referred to as glycogen, to the treatment en bloc with uranyl acetate. Material was fixed in glutaraldehyde and OsO4. Blocks were processed either conventionally or immersed in uranyl acetate before dehydration. Sections were examined unsatined, stained with U and/or Pb or with a histochemical technique (PA-TSC-SP) specific for glycogen. Electron dense granules have affinity to Os, U and Pb which suggests ionic reactions specific for protein but improbable for glycogen. Large granules in A turned into pale ghosts and small granules in B disappeared after treatment en bloc with uranyl acetate. PA-TSC-SP in conventional samples showed glycogen particles arranged into aggregates corresponding in size to the electron dense granules. Treatment en bloc slightly affected glycogen aggregates in A and resulted in a formation of large clumps of glycogen particles in B. It was concluded that the electron dense granules represented protein bound to glycogen in the organelles called glycosomes. Acidic action of uranyl acetate removed protein from glycosomes. The degree of this removal depended on the amount of protein present in glycosomes in the moment of fixation.


Production of antibodies to human immunoglobulins in a gibbon. Immunization of a chimpanzee, a gibbon, a baboon and a rhesus monkey with human globulins gave different results. While chimpanzees did not produce any anti-human antibodies, baboon and rhesus monkey responded by producing anti-human IgG antibodies that did not show subclass or allotype specificity. In the gibbon antiserum, on the other hand, six different antibodies were demonstrated with a specificity against one or more of the four human IgG subclasses, including one allotype. Comparative investigations of non-human primate species sera showed that one of the specificities detected by these antibodies, i.e., the allotype G1m(f), was restricted to man; the other five antibodies reacted with man and with two or more genera of apes. Lower primates did not share any of these antigenic determinants. The extent of immunological distance is parallel to the taxonomic classification. The gibbon shows that there are more similarities between man and chimpanzee or pygmy chimpanzee than between man and gorilla or orangutan.


In vitro comparison of the erythrocyte sedimenting properties of dextran, hydroxyethyl starch and a new low-molecular-weight hydroxyethyl starch. A low-molecular-weight preparation of hydroxyethyl starch (LMW-HES) may serve as a desirable substitute for the erythrocyte sedimenting agents presently employed to improve neutrophil yields in centrifugation and gravity leukapheresis. The cell-separating and erythrocyte-sedimenting properties of LMW-HES, assessed in vitro by the recovery of various blood cells in plasma supernatant fluids, were similar to those of the higher-molecular-weight hydroxyethyl starch in current use and to dextran under a variety of conditions. These data predict success for LMW-HES as a sedimenting agent in leukapheresis and recommend that it be evaluated in clinical trials.


[Short-term use of lithium carbonate in the treatment of thyrotoxicosis]. The examinations were performed on 3 groups of altogether 65 persons with thyreotoxicosis, the cause of which was either Basedow's disease or struma nodosa. The first group received metizol (thiamazol) in a daily dosage of 60 mg, the second group lithium carbonate (1.0 up to 1.5 g/a day), the third group metizol together with lithium carbonate. The groups were of the same value, concerning the degree of the symptoms of hyperthyroidism. The examinations showed that when lithium carbonate is used alone at the earliest a significant decrease of the serum T4 and T3 concentration as well as of the T3 binding index appears. After a treatment of seven days the therapeutic effects even up. Under the lithium therapy an essential improvement of the clinical findings was achieved. The combined therapy with lithium and metizol did not exhibit any advantages in this respect. The side-effects observed under the lithium therapy are no essential clinical problem.


[Monoreceptor sera to the antigenic determinants of influenza virus A hemagglutinin]. The method of specific adsorption was used to produce monoreceptor sera to individual antigenic determinants of hemagglutinin of influenza A virus, Hong Kong variant. The sera were completely free of inhibitors, contained no antibody to neuraminidase or host cell antigens. The monoreceptor sera were used in HI tests and immunoelectrophoresis for the qualitative and quantitative analysis of the antigenic variants isolated in 1968--1977. It was shown that alongside with the appearance of qualitatively new antigenic determinants changes in the content of some or other antigens in influenza virus hemagglutinins occurred.


Identification and characterization of some serum proteins of the laboratory rat (Rattus norvegicus) by crossed immunoelectrophoresis. Rat (Rattus norvegicus) serum proteins were examined by means of crossed immunoelectrophoresis. We have performed the topographic characterization of prealbumins, albumin, orosomucoid, glycoprotein of Darcy, alpha-macroglobulins, haptoglobin, hemopexin, transferrin and IgG. There are several differences in rat serum in comparison to human serum. This applies, for example, to prealbunins and to Darcy's glycoprotein. The behaviour of these fractions in different conditions is discussed in more detail.


5 years' experience of 5-S-cysteinyldopa in melanoma diagnosis. Determinations of the urinary excretion of 5-S-cysteinyldopa were performed in 571 patients previously treated by surgery for melanoma or melanoma metastasis. 90% of the 161 patients with metastases showed values exceeding 0.15 mg/24 h, and 9% of the 410 patients without metastases had such values. The increase in 5-S-cysteinyldopa excretion was generally more pronounced in men with metastases than in women, 98% of the men and 77% of the women with metastases showing values exceeding 0.15 mg/24 h. High levels of 5-S-cysteinyldopa are of grave prognostic significan4% died within one month, and only 3% survived for more than a year. In Sweden, determination of 5-S-cysteinyldopa in patients operated on for melanoma gives maximum information in the winter (October--March), when sun exposure does not influence the excretion levels.