The significance of a differential distribution of phosphomonoesterases on bone surfaces after prolonged demineralization. An observed differential distribution of alkaline and acid phosphatase on the surfaces of growing bones may serve to describe transformative processes of bone growth. This conclusion has been reached by comparing the distribution of the two enzymes on the surfaces of fibulae from young rats with the patterns of apposition and resorption on the periosteal surfaces of this bone, revealed by in vivo staining with alizarin red S. Presence of reaction to acid phosphatase is, as shown before, an indication of resorptive surfaces, while the presence of reaction to alkaline phosphatase is an indication of depository surfaces.


The histochemical localisation of aminopeptidases in the central nervous system and an analysis of factors contributing to the final staining pattern. A method is described for the localisation of aminopeptidases in the central nervous system. The enzymatic cleavage of the peptide bond in the substituted naphthylamide substrates results in the liberation of free naphthylamine which in the presence of an azo salt can be precipitated as a blue dye at sites of enzyme activity. Analysis of different brain regions using this technique indicates that these enzymes may have a specific function at certain sites in the CNS. Since this is the first method to produce any localised staining in nervous tissue an analysis of factors contributing to the final staining pattern is also presented.


The zinc iodide-osmium tetroxide staining-fixative of Maillet. Nature of the precipitate studied by x-ray microanalysis and detection of Ca2+-affinity subcellular sites in a tonic smooth muscle. A mechanism of osmium reduction during zinc iodide-osmium tetroxide (ZIO) fixation is proposed. X-ray powder microanalyses of ZIO precipitates formed both in the presence or absence of tissues are identical with those of CuOsO4 and CuRuO4. Therefore, and based on indexation methods, ZnOsO4 was found to be the formula of the ZIO mixture reduction; this zinc osmate has an orthorhombic crystalline lattice. In smooth muscle preparations, ZIO electron dense deposits are localized in both cisternae of the sarcoplasmic reticulum and in mitochondria after a short fixation time. According to the microanalysis results, the zinc osmate has been associated to Ca2+ high affinity sites since Zn2+ is either replacing Ca2+ and/or displacing it by having a higher affinity for Ca2+ binding sites. Consequently, the ZIO mixture might be useful in revealing some Ca2+ storage sites in cells. This hypothesis was tested in ABRM preparations by selectively depleting sites which are known to bind Ca2+. In this case, the sarcoplasmic reticulum only retains the staining deposits after a short ZIO fixation. It is likely that OsO4 alone, used as fixative in cytology might be due to the formation of metallic osmates (e.g., divalent osmates like CaOsO4). In addition, of course, reduction of osmium during tissue fixation is accompanied by oxidation of double bonds of lipoproteic complexes or unsaturated lipids, and oxidation of sulfhydryl groups and amino groups.


Multi-media graphics: a transparency system. Persons responsible for teaching in the health sciences are justifiably concerned about the cost of their visual presentations. These graphics can be used as slides and in publications and illustrators need to know how the transparency system can convert publication material into effective color visuals as 35mm slides, video tapes or television graphics at a moderate cost.


A rat serum glycoprotein whose synthesis rate increases greatly during inflammation. Crossed immunoelectrophoresis of rat serum demonstrated considerably increased serum concentrations of at least ten different proteins during turpentine-induced inflammation. One protein, which moved during electrophoresis like an alpha 1 globulin, showed a particularly large increase. This protein was purified to homogeneity by ammonium sulfate fractionation followed by chromatography on DEAE-cellulose. Sephadex G-100, and concanavalin A-Sepharose, and finally disc electrophoresis in polyacrylamide gel. It has a molecular weight of 56,000 determined by equilibrium ultracentrifugation. An apparent molecular weight of 68,000 was estimated for the reduced protein by electrophoresis in polyacrylamide gel plus sodium dodecyl sulfate, suggesting that the native protein is composed of a single polypeptide chain. It has an E2801%, 1 cm of 5.2, an isoelectric pH of 4.7, and contains 19% carbohydrate. The protein does not inhibit bovine trypsin or chymotrypsin. Its physical properties and amino acid composition distinguish this protein from all other rat serum proteins hitherto characterized. During acute inflammation, induced 25 h previously, rats incorporated 20 times more [14C]leucine into this particular protein than did normal rats. However, incorporation into total serum protein during acute inflammation increased only slightly. Regardless of whether inflammation was induced by surgical injury or by a subcutaneous turpentine injection, within 48 h the serum concentration of this major acute-phase protein rose from the normal value of 0.46 g/liter to a maximum value of 7.2 g/liter, which constituted 10% of the total serum protein.


The staining pattern of collagen fibrils. Improved correlation with sequence data. The periodic banding pattern of stained collagen fibrils observed in the electron microscopic can be correlated with the charge distribution deduced from the amino acid sequence. Earlier work used alpha 1 chain sequence data only. The present study incorporates alpha 2 as well as alpha 1 sequence data, so that the complete distribution of charged residues is used. Correlation is improved if it is supposed that the extrahelical terminal regions are contracted. The optimal value of the periodicity, D, (previously 232.3 +/- 0.5 residues using alpha 1 data only), is now 234.2 +/- 0.5 residues, assuming uniform spacing of residues in the helical body of the molecule. This value agrees better with values obtained by others from analyses of interactions between molecules, using sequence data alone. Using the improved value of D, the relative axial locations of the charged residues in the fibril are displayed. In this way, the charged residues contributing to each band in the fibril staining pattern can be identified.


Iron-bleomycin interactions with oxygen and oxygen analogues. Effects on spectra and drug activity. Despite extensive structural dissimilarities, iron . bleomycin complexes and heme-containing oxygenases display remarkable similarities in binding oxygen antagonists and in spectral properties deriving from bound iron. Fe(II)-bleomycin reversibly forms a complex with either CO or isocyanide (lambda max = 384 and 497 nm, respectively), either of which interfere with its oxygen-dependent cleavage of DNA. A similar but paramagnetic complex forms with NO (lambda max = 470 nm; AN = 24 G). In contrast, cyanide enhances bleomycin activity against DNA. Complexes of bleomycin and FE(III), formed either by direct association or by autoxidation of the Fe(II) . bleomycin complex, exhibit indistinguishable EPR and visible spectra, which change characteristically with pH. At neutral pH, Fe(III) . bleomycin is a low spin complex (g = 2.45, 2.18, 1.89; lambda max = 365, 384 nm) and, at low pH, it is a high spin rhombic complex (geff = 9.4, 4.3; lambda max = 430 nm). These complexes are interconvertible (pK 4.3). Fe(II) . bleomycin oxidation, although reversible by spectral criteria, is accompanied by drug inactivation unless DNA is present.


The effects of thioureylene compounds (goitrogens) on lactoperoxidase activity. The rates of oxidation of several goitrogens by lactoperoxidase and the rates of inactivation of lactoperoxidase by the same goitrogens have been measured. The influence of iodide on both reactions has also been evaluated. It has been shown by us that iodide acts catalytically in regulating lactoperoxidase activity at pH 8.8. The rate data have been analyzed by a computer program which solves the differential equations for the above mentioned reactions. From this computer analysis we have been able to obtain binding constants of the goitrogens and inactivation rate constants of lactoperoxidase. Iodide was shown to inhibit goitrogenic activity either by increasing the rate of drug oxidation or by reducing the rate of enzyme inactivation, or both, depending on the particular drug. Iodide had little or no effect on the goitrogen-binding constants. We have also shown that the relative rates of enzyme inactivation can be correlated with the potency of the goitrogen as an antithyroid drug.


Is Na+-dependent exchange diffusion a true exchange? Trans-stimulation of glycine uptake by cellular glycine in Ehrlich cells is a Na+-dependent phenomenon. In contrast trans-stimulated methionine or leucine uptake is Na+-independent. Trans-stimulated uptake of glycine does not show any characteristics of an ex change process but rather appears to be due to changes in membrane potential which occur as a result of a net Na+-dependent loss of cellular amino acids. Trans-stimulated influx of glycine occurs during the time of net loss of cellular glycine and is absent when the cellular amino acid level is at steady or when the cell is depolarized. Exchange of leucine or methionine occurs when the amino acid level is at steady state and it is not directly affected by depolarizing agents such as gramicidin.


Some properties of polyethylene glycol:phenylalanine ammonia-lyase adducts. Methoxypolyethylene glycol of 5000 daltons (PEG) was attached covalently to phenylalanine ammonia-lyase from Rhodotorula glutinis. Attachment of sufficient quantities of PEG to phenylalanine ammonia-lyase substantially reduces immunological recognition and clearance of the conjugated enzyme in mice. The modified enzyme demonstrates altered catalytic properties such as shifts in the pH and temperature optima, an increase in the Michaelis-Menten constant, and a lowered Vmax in comparison with the native enzyme. PEG-phenylalanine ammonia-lyase has increased resistance to proteolytic digestion, particularly when in the presence of cinnamate, a competitive inhibitor, while the native enzyme is rapidly inactivated. In the ultracentrifuge PEG-phenylalanine ammonia-lyase exhibits a lower sedimentation rate than the unmodified enzyme, despite the fact that it is much larger. The electrophoretic mobility of PEG-phenylalanine ammonia-lyase is greatly decreased in comparison to the unmodified enzyme. PEG-phenylalanine ammonia-lyase had a much longer blood-circulating life in mice, both initially and after a number of injections, than did the native enzyme. PEG-phenylalanine ammonia-lyase was a good immunogen but a poor antigen in mice and rabbits, that is, it readily induced antibody formation, but reacted poorly in vitro with the antibodies that were formed against it.


Human alpha-fetoprotein. Fluorescence studies on binding and proximity relationships for fatty acids and bilirubin. The binding of bilirubin and the polyene fatty acids cis-parinaric acid and cis-eleostearic acid to human alpha-fetoprotein was studied using fluorescence quenching and fluorescence enhancement techniques. alpha-Fetoprotein has three fatty acid binding sites of decreasing affinity (association constants 2.1 x 10(7) M-1 9.1 X 10(5) M-1, and 1.4 x 10(5) M-1) and one relatively strong and one relatively weak bilirubin binding site (association constants 1.1 x 10(7) M-1 and 1.8 x 10(5) M-1). These association constants are slightly weaker than the corresponding association constants for binding to human albumin. Competition experiments failed to show preferential binding of polyunsaturated fatty acids. Fluorescence quenching was used to determine 11 ligand-ligand and ligand-tryptophanyl residue distances. Each of these 11 calculated distances (ranging from 19 A to 32 A) was within 5 A of the corresponding distances measured previously for human albumin (Berde, C.B., Hudson, B.S., Simoni, R.D., and Sklar, L.A. 1979, J. Biol. Chem. 254, 391-400). Thus, in addition to previously described sequence homology, immunologic cross-reactivity, and other similarities, human albumin and human alpha-fetoprotein have functional and geometric homologies.


Topographic antigenic determinants on cytochrome c. Immunoadsorbent separation of the rabbit antibody populations directed against horse cytochrome. Seven populations of site-specific antibodies were isolated from each of three sera of rabbits immunized against glutaraldehyde-polymerized horse cytochrome c. The antibodies were separated using an immunoadsorption scheme which employed the following cytochromes c: horse, beef, guanaco, rabbit, mouse testicular, pigeon, and the cyanogen-bromide cleaved fragment of the rabbit protein containing residues 1 to 65. The monovalent, antigen-binding fragments of the antibodies (Fab') gave 1:1 stoichiometries with native horse cytochrome c in fluorescence quenching assays. Cross-reactivities with heterologous cytochromes c using fluorescence quenching and a modified Farr assay demonstrated that the antigenic determinants are situated around residues 44, 60, and 89/92, four of the six amino acid sequence positions where horse and rabbit cytochromes c differ. The remaining two differences occur at residues 47 and 62. The apparent lack of immunogenicity of these two substitutions may result from the presence of the more immunogenic residues 44 and 60 nearby. Of the seven antibody populations isolated, four were shown to bind in the region of residues 89 and 92. Since several cytochromes c have amino acid sequence differences from the horse protein at either of these two residue positions, it was possible to fractionate the antibodies directed against this complex site on the basis of subtle specificity differences between them. Two antibody populations bind in the region of residue 44. One of these is specific for proline at that position, while the other antibody population also binds to cytochrome c containing glutamic acid at position 44. The remaining antibody population binds in the region of the lysine residue at position 60. Each of the seven site-specific antibody populations binds effectively to any cytochrome c having a suitable amino acid sequence in the antigenic determinant regardless of any residue differences from the immunogen outside of that area. It was also demonstrated that these seven antibody populations represent the totality of the antibodies elicited in rabbits against horse cytochrome c, since the immunoadsorbants bound all the antibodies specific for the native protein. Furthermore, the rabbit antisera contained no other antibody population that could bind to the conformationally disturbed, cyanogen bromide-cleaved fragment of horse cytochrome c containing residues 1 to 65, making it appear that there were no antibodies elicited against a "processed" form of cytochrome c.


Oxygenated iron bleomycin. A short-lived intermediate in the reaction of ferrous bleomycin with O2. The reaction of Fe(II) . bleomycin with O2 to yield Fe(III) . bleomycin has been resolved into two kinetic events by stopped-flow spectrophotometry. The first event is first order with respect to both bleomycin and O2 and may be regarded as a second order reaction (k = 6.1 x 10(3) M-1s-1 at 2 degrees C). The first product has no EPR spectrum. The optical spectrum resembles those of Fe(II) . bleomycin complexes with CO, NO, and ethyl isocyanide. We propose that the first product is an Fe(II) . bleomycin . O2 complex. The second kinetic event is first order with respect to the first accumulated product (k = 0.11 s-1 at 2 degrees C) and independent of oxygen concentration. The product of this reaction is indistinguishable from Fe(III) . bleomycin by optical and EPR spectroscopy.


Chemotherapy studies in autochthonous rat tumors: carcinomas of the forestomach. Tumors of the forestomach were induced in male Sprague-Dawley rats by oral application of acetoxymethyl-methylnitrosamine (AMMN) in single weekly doses of 3.5 mg/kg body weight for 10 weeks. 100 +/- 5 days after the beginning of carcinogen treatment exploratory laparotomy was performed. One hundred animals in the same stage of tumor development were divided at random into one control group and four groups treated with cytostatics. Bleomycin (BLM), methotrexate (MTX), 5-fluorouracil (5FU), and 1,2-bis-(2-chloroethyl)-1-nitrosourea (BCNU) were tested in groups of 20 rats each. Only in animals receiving repeated doses of BLM a slight tumor response was observed but no increase in median survival times was seen. No therapeutic effects of the other drugs used were demonstrable.


Carboxyhaemoglobin levels in workers in Leicestershire garages. Blood carboxyhaemoglobin (HbCO%) levels were measured in 61 workers in 35 garages in Leicester and Leicestershire. Of 26 workers in large garages in winter, 44% of the non-smokers and 20% of the smokers had increased CO absorption, the highest HbCO% being 26.0 in a non-smoker. Of 35 workers in 24 smaller, randomly chosen garages visited throughout the year (two per month) the corresponding figures were 43% and 14%, the highest HbCO% being 21.0. The possible relevance of these findings to coronary heart disease in garage workers is discussed.


Antibodies that bind specifically to synaptic sites on muscle fiber basal lamina. Basal lamina (BL) ensheathes each skeletal muscle fiber and passes through the synaptic cleft at the neuromuscular junction. Synaptic portions of the BL are known to play important roles in the formation, function, and maintenance of the neuromuscular junction. Here we demonstrate molecular differences between synaptic and extrasynaptic BL. We obtained antisera to immunogens that might be derived from or share determinants with muscle fiber BL, and used immunohistochemical techniques to study the binding of antibodies to rat skeletal muscle. Four antisera contained antibodies that distinguished synaptic from extrasynaptic portions of the muscle fiber's surface. They were anti-anterior lens capsule, anti-acetylcholinesterase, anti-lens capsule collagen, and anti-muscle basement membrane collagen; the last two sera were selective only after antibodies binding to extrasynaptic areas had been removed by adsorption with connective tissue from endplate-free regions of muscle. Synaptic antigens revealed by each of the four sera were present on the external cell surface and persisted after removal of nerve terminal. Schwann cell, and postsynaptic plasma membrane. Thus, the antigens are contained in or connected to BL of the synaptic cleft. Details of staining patterns, differential susceptibility of antigens to proteolysis, and adsorption experiments showed that the antibodies define at least three different determinants that are present in synaptic but not extrasynaptic BL.


Rapid presumptive bacteriological diagnosis of Legionnaires disease. A simple, relatively rapid silver impregnation stain has been found to stain Legionella pneumophila effectively in paraffin-embedded tissue sections while permitting visualization of histological detail. It may also be used to stain the organism in body fluids. The stain is not specific and thus must be confirmed by direct fluorescent-antibody technique or culture, but, in the absence of other bacilli demonstrable by Gram or other stain, visualization of typical bacillary forms in a patient with illness compatible with Legionnaires disease provides strong presumptive evidence supporting this diagnosis.


Effect of various histological fixatives on fluorescent antibody detection of Legionnaires disease bacteria. Human lung tissue containing the Legionnaires disease bacterium was fixed in seven different histological fixatives, processed, and embedded in paraffin. Deparaffined sections from each were stained by fluorescent antibody and by Dieterle silver impregnation. With the fluorescent antibody stain, the Legionnaires disease bacterium could be detected in tissues prepared with any of the fixatures, but the Dieterle silver impregnation was not satisfactory on Zenker-fixed tissues.


Immunochemical evidence for multiple serotypes of Bacteroides fragilis. An immunochemical comparison of outer membrane antigens obtained from five select and biochemically defined strains indicated that there are several serotypes of Bacteroides fragilis. Each strain was serologically defined by individual or by combinations of determinant groups composed of carbohydrates in the form of polysaccharides or glycoproteins. The carbohydrate constituents were tentatively identified as glucose, galactose, fucose, rhamnose, glucosamine, galactosamine, and traces of mannose. Strains were observed to have minor qualitative and major quantitative variations in carbohydrate composition.


Detection of antilymphocyte antibody with two-color method in systemic lupus erythematosus and its heterogeneous specificities against human T-cell subsets. The two-color method originally described by Van Rood et al. (Van Rood, J. J., A. Van Leeuwen, and J. S. Ploen. 1976. Simultaneous detection of two cell populations by two-color fluorescence and application to the recognition of B-cell determinants. Nature (Lond.). 262: 795-797) for the typing of homologous leukocytic antibodies, D-region was used for the detection of antilymphocyte antibody (ALA) in systemic lupus erythematosus. In this method, surface immunoglobulin-bearing cells were identified with fluorescein isothiocyanate-labeled anti-immunoglobulin and nuclei of killed cells were stained with ethidium bromide. Therefore, cell type (T or B) of the target cells can be identified without fractionating them. ALA was detected in 87% of lupus sera and had a preferential reactivity with T cells. Its major immunoglobulin class was shown to be immunoglobulin (Ig)M. The subspecificity of ALA was further analyzed using fractionated T-cell subsets as target cells. When T lymphocytes were separated into Fc receptor-bearing (Tgamma) and lacking (Tgamma[-]) cells, 64% of ALA showed preferential reactivity with Tgamma cells and 14% with Tgamma(-) cells. The remainder had no selective reactivity against Tgamma or Tgamma(-) cells. Tgamma cells were shown to have suppressor activity, whereas Tgamma(-) cells were indicated to contain helper cells. The above finding was in agreement with the observation that treatment of T cells with ALA that preferentially react with Tgamma cells considerably enhanced immunoglobulin synthesis in vitro, whereas treatment of T cells with ALA reactive with Tgamma(-) cells clearly suppressed the formation of immunoglobulins. Treatment of ALA with no selective reactivity showed variable effects on in vitro immunoglobulin synthesis. These results indicate that ALA in lupus have heterogeneous specificities against human T-cell subsets.


Generation of a fibroblast chemotactic factor in serum by activation of complement. When serum complement is activated by either the classical or alternative pathways, a factor with an apparent 80,000 mol wt is generated that is chemotactic for human dermal fibroblasts. The origin of this serum-derived chemotactic factor (SDCF) is not known; however, it may be a cleavage product from C5 because it is inactivated by monospecific antiserum to human C5, and it is not generated when the complement system is activated in human serum deficient in C5. SDCF is not chemotactic for human neutrophils or monocytes. Because SDCF is generated when serum complement is activated, it may function in vivo to attract connective tissue fibroblasts to sites of inflammatory reactions in which the complement system participates.


Hypothyroidism as a late sequela in patient with Graves' disease treated with antithyroid agents. In 1971, thyroid function was evaluated in 15 unselected patients whose only therapy for diffuse toxic goiter was a course of thionamide drug treatment completed 20-27 yr earlier. One patient was frankly hypothyroid by clinical and laboratory criteria. The remaining 14 patients appeared clinically euthyroid and had a normal serum thyroxine (T(4)) concentration and thyroid radioiodine uptake (RAIU). Nevertheless, only 6 of 14 appeared to be entirely normal according to more refined criteria. The serum thyrotropin (TSH) concentration was markedly elevated in one patient and above the normal range (1.6+/-2.0; mean+/-2 SD) in five others. Thyroid stimulation with exogenous TSH revealed subnormal responses of the serum T(4)I, RAIU, or both, in 7 of 11 patients tested. An abnormal iodideperchlorate discharge test was found in 5 of 10 patients and appeared most abnormal in patients with abnormal RAIU responses to TSH. Fluorescent antimicrosomal antibody was found in the serum of 12 of the 15 patients, in contrast to an expected frequency of 7% in normal individuals of the same age. By the time a second major follow-up study was completed in 1978, two additional patients had become frankly hypothyroid. A third subject met accepted criteria for subclinical hypothyroidism. One of these subjects had had a clearly elevated serum TSH concentration in 1972, and the remaining two had exhibited the highest responses of serum TSH (36, 26 muU/ml) to thyrotropin-releasing hormone among 10 patients tested in 1972. One patient developed recurrent thyrotoxicosis in 1978, 25 yr after the onset of his first and only other apparent episode of hyperthyroidism. This patient was the only one who demonstrated a subnormal response to thyrotropin-releasing hormone in 1972. The remaining nine subjects that could be studied in 1978 exhibited varying combinations of abnormalities of thyroid function. It is concluded that progressive failure of thyroid function is a common occurence in long-standing Graves' disease, and it is suggested that this results from concomitant chronic thyroiditis. We postulate that this inherent tendency toward thyroid failure is exaggerated by surgery or radioactive iodine, explaining the progressive increase in, and inordinate frequency of, hypothyroidism after ablative modes of therapy in diffuse toxic goiter.


Cross-idiotypic antigens among monoclonal immunoglobulin M from patients with Waldenström's macroglobulinemia and polyneuropathy. The monoclonal immunoglobulin (Ig)M from 5 to 16 patients with Waldenström's macroglobulinemia and a polyneuropathy shared cross-idiotypic antigenic determinants as demonstrated by hemagglutination and hemagglutination inhibition experiments as well as by precipitin reactions. This reactivity was located to the Fab (and not Fc) fragment of the protein. The IgM from 73 patients with macroglobulinemia but without neuropathy all gave negative reactions. In contrast, the monoclonal IgG from a patient with polyneuropathy also possessed similar idiotypic determinants. Since cross-idiotypic determinants are usually related to the combining site of a monoclonal Ig, this finding suggests that the monoclonal Ig of these patients may mediate the nerve injury via their antibody activity, which could be directed either to a nerve antigen or to some component involved in the pathogenesis of the neuropathy.


Human endodermal sinus tumour in nude mice and its markers for diagnosis and management. Two human endodermal sinus tumours (yolk sac tumours) were transplanted successfully into nude mice. The transplanted tumours maintained not only morphological characters, such as Schiller-Duval bodies, but also the ability to synthesise alpha-fetoprotein, lactic dehydrogenase 1, liver and bone type alkaline phosphatase, and some human serum proteins. Since these tumours produced lactic dehydrogenase 1 but not the other four isozymes of lactic dehydrogenase, this isozyme, like alpha-fetoprotein, seems to be a good marker for the diagnosis and management of cases of endodermal sinus tumour. One of the two tumours produced another fetal antigen or carcinoembryonic antigen in addition to alpha-fetoprotein. These two endodermal sinus tumours, with their various markers in nude mice, will be useful in studies on diagnostic markers.


A comparative study of the immunohistochemical localization of basic protein to myelin and oligodendrocytes in rat and chicken brain. Antisera to highly purified basic protein (BP) from rat and chicken brain were prepared and their purity and specificity demonstrated by double immunodiffusion and cross-immunoadsorption. These antisera were used for immunohistochemical localization of BP in the brains of adult and developing rat and chick. Myelin basic protein was exclusively localized to myelin or the myelin forming elements of the CNS. It was present in high concentrations in white matter and absent in areas free of myelin. Neuronal parikarya and dendrites were negative as were axons cut in cross section and at Nodes of Ranvier. The latter was best observed in cross sections of human spinal cord demonstrating also the immunoreactivity of the antibodies with human BP. The internodal distance in a fine (1.5 micrometer) rat cortical fiber was determined to be approximately 45 micrometers. Myelin basic protein was shown to extend into cranial roots, in contrast to myelin proteolipid protein which abruptly lose fluorescence as the nerves emerged from the brain. During development, BP was first observed on the fourteenth day of incubation in chick and at birth in the rat. The protein appeared in oligodendrocytes and in association with fibers near these cells. Fluorescent processes were frequently observed connecting the oligodendrocytes with the fibers. As myelination progressed, the intensity of the immunohistochemical reaction decreased in the oligodendrocytes while the brightness in fibers increase. Eventually, the oligodendrocytes became undetectable. Fibers with immature myelin exhibited a beaded or varicosed appearance with the highest concentration of immunofluorescence in the outer portion of the varicosities. The varicosities were postulated to represent dilations in the newly forming sheath between intervals of compaction along the axon undergoin myelination. These dilations might represent areas of increased cytoplasmic volume which could serve as channels for transport and/or storage sites for myelin proteins prior to incorporation into the membrane. The varicosities became less prominent with the thickening of the myelin sheath and mature myelinated fibers became smooth. The process of synthesis of BP, transport of the protein to the varicosed fibers, and maturation of the myelin sheath was seen to progress in a more or less caudal to rostral direction as myelination of the CNS takes place. In the rat, this was accomplished over approximately a 30-day period starting near the time of birth. In the chick, most of the myelination was accomplished in the three or four days immediately before hatching. At this time, innumerable oligodendrocytes were observed producing BP simultaneously in the major white fiber tracts. It is postulated that in chick some degree of oligodendrocytic cell death occurs normally during myelination.


Effects of delta 1-tetrahydrocannabinol on cyclic AMP in cultured human diploid fibroblasts. (-)-trans-delta 1-Tetrahydrocannabinol (delta 1-THC) antagonized the cyclic AMP responses of WI-38 fibroblasts to both prostaglandin E1 (PGE1) and catecholamines. Both cellular cyclic AMP accumulation and cyclic AMP escape to the incubation medium were reduced, but the reduction of escape was much more dramatic at all concentrations of the drug. Conversely, long term incubations of cells with delta 1-THC alone resulted in substantial accumulations of cyclic AMP in the incubation medium. This effect was potentiated by the phosphodiesterase inhibitor 1-methyl, 3-isobutylxanthine and appeared to result from weak agonist activity of the cannabinoid as determined by a) stimulation of radioactivity incorporated into cyclic AMP using 3H-adenine prelabelled cells, and b) a rapid and pronounced increase in the activity ratio of cellular protein kinase. The antagonistic effect of delta 1-THC on the cellular response to PGE1 was greater in preconfluent cells than in confluent monolayers. Further, the increased sensitivity of preconfluent cultures to delta 1-THC was associated with the appearance of cytoplasmic vacuoles in the perinuclear region of the cells. Cannabidiol acted similar to delta 1-Thc in affecting cyclic AMP metabolis whereas cannabinol and cannabicyclol showed mixed effects on the various parameters studied.


Immunologic tolerance to allergenic protein determinants: properties of tolerance induced in mice treated with conjugates of protein and a synthetic copolymer of D-glutamic acid and D-lysine (D-GL). Conjugates of proteins and the synthetic copolymer of D-glutamic acid and D-lysine (protein-D-GL) reproducibly induce significant unresponsiveness to the protein antigens in experimental mice. Proteins studied include ovalbumin and antigen E of ragweed extract, the major allergen of ragweed pollen. The unresponsive state 1) can be induced in both unsensitized and previously sensitized experimental animals, 2) is selectively confined to responses of the IgE antibody class, 3) is long lasting, and 4) is highly antigen specific. IgE antibody responses can also be suppressed by administering comparable doses of unconjugated protein alone; however, the unresponsive state induced in this manner is only transient and rebound production of IgE antibody is often observed. Results from the studies of the cellular basis of the protein-D-GL induced unresponsiveness demonstrate that 1) protein-D-GL conjugates do not induce unresponsiveness at the level of protein-specific B cells, 2) tolerance is not induced by virtue of a detectable antigen-specific suppressor T cell mechanism, 3) tolerance is most probably induced in the antigen-specific helper T cell populations. The significant IgE-selective and antigen-specific tolerogenic activity of protein-D-GL conjugates make these compounds potential candidates for use as therapeutic agents in the treatment of IgE-mediated human allergic disorders induced by protein allergens.


Propagation of antigen-specific T cell helper function in vitro. Antigen-induced proliferation of primed lymph node cells was abrogated by treatment with anti-Ly 1 serum and complement (C) but not with anti-Ly 2 serum and C. Lymph node cells from animals primed to ovalbumin were activated with antigen in vitro, followed by propagation in an antigen-free supernatant fluid obtained from lectin-induced normal spleen cells. T cells processed in this manner displayed a stepwise enrichment of helper activity for antibody production as measured in a secondary hapten-carrier response. The sequential increase in antigen-specific help was paralleled by a rise in the antigen-induced proliferative response, a phenomenon whose expression was dependent on the presence of syngeneic or semi-syngeneic irradiated filler cells.


The murine Kupffer cell. I. Characterization of the cell serving accessory function in antigen-specific T cell proliferation. Murine Kupffer cells, the tissue macrophages of the liver, were isolated by collagenase digestion, differential sedimentation over Metrizamide, and glass adherence. The resultant cell population was more than 86% phagocytic, and 95% of cells stained positively for alpha-naphthyl butyrate esterase activity. The cells also had cell surface receptors for complement (C) and the Fc portion of IgG. In addition, a large proportion of Kupffer cells was shown to bear Ia antigens: about half of the cells bore I-A subregion-encoded antigens and about half bore I-BJE or I-EC subregion-encoded antigens. Kupffer cell populations were capable of reconstituting antigen-stimulated proliferative responses of antigen-primed, macrophage-depleted, lymph node T cells. The ability to reconstitute proliferation was enriched in the adherent population and was resistant to radiation and treatment with an anti-Thy antiserum and C. We conclude that isolated murine Kupffer cells bear the Ia phenotype of accessory cells that function in antigen presentation and that Kupffer cells can participate in the induction of antigen-specific immune responses. These data suggest that Kupffer cells may play a role in modulating responses to enterically derived antigens.


The specificity of human autoantibodies that react with both cell nuclei and plasma membranes: the nuclear antigen is present on core mononucleosomes. We have examined the nature of the nuclear antigen recognized by certain natural human antibodies that react specifically with both cell nuclei and plasma membranes from many species. Partial purification of these antibodies, called X-ANA, is achieved by binding to and rapid elution from the surface of viable human leukocytes. Chicken erythrocyte chromatin was solubilized by digestion with staphylococcal nuclease and fractionated into a 0.15 M NaCl soluble fraction that consisted of core mononucleosomes lacking H1/H5, and a 0.15 M NaCl insoluble fraction composed of polynucleosomes with H1/H5 present. No proteins other than histones were detected. Native and reconstituted mononucleosomes displaced IgG of the leukocyte eluates from nuclei of frozen mouse kidney sections and from the walls of plastic tubes coated with polynucleosomes. The reconstituted core mononucleosomes were 4- 10-fold less efficient inhibitors than native mononucleosomes. Trypsin digested mononucleosomes, free high m.w. DNA, and free histones displayed no or very weak inhibitory activity. The data indicate that X-ANA recognize a complex consisting of the core histones H2A, H2B, H3, H4, and DNA of 140 to 200 base pairs in length.


Role of nominal antigen and Ia antigen in the binding of antigen-specific T lymphocytes to macrophages. We have previously demonstrated that when primed T lymphocytes were repeatedly incubated on monolayers of antigen-pulsed macrophages (M phi), the cells that failed to adhere to the monolayer demonstrated a marked depletion of their proliferative response that was specific both for the antigen used for pulsing the M phi and for Ia determinants on the M phi. In order to further analyze the contribution of the nominal antigen and Ia antigens to the physical binding of T lymphocytes to M phi, we have attempted to block the absorption of T lymphocytes to M phi with a large excess of soluble antigen and with anti-Ia sera. Our results demonstrate that anti-Ia sera inhibit but that soluble antigen augments the binding of specific T lymphocytes to M phi. The implications of these findings for "dual recognition" and "linked recognition" models of T lymphocyte receptors are discussed.


Use of immunoadsorbents for the study of antibody binding to sperm whale myoglobin and its synthetic antigenic sites. Conditions for preparing immunoadsorbents of sperm-whale myoglobin and its five synthetic antigenic sites and for desorption of radiolabeled antibodies from the immunoadsorbents were studied. In immunoadsorbent titration studies, the sum of the amounts of antibodies bound in the plateau (maximum binding) by the adsorbents of the five sites accounted quantitatively for the entire (100%) antibody response to sperm-whale myoglobin.


The isolation of an optimal small size immunoferritin conjugate fraction. A procedure is described for preparing immunoferritin conjugates consisting of one antibody molecule attached to one ferritin molecule. Results obtained with such a reagent faithfully reflect the true distribution of antigenic determinants, and allow more accurate quantification.


The immunoglobulin-like T-cell receptor. I. In situ demonstration of immunoglobulin Fab-region determinants of rodent T- and B-lymphocytes using chicken antibodies. Antibodies were raised in chickens to the (Fab')2 fragment of normal murine IgG and to the k-myeloma protein MOPC 41. Following appropriate absorptions or purification by immune affinity chromatography the chicken antibodies bound specifically to both T- and B-cells of mice, rats and guinea-pigs as detected by quantitative cytofluorescence, radioactive binding assays and transmission and scanning immunoelectronmicroscopy. These antibodies are directed against polypeptide determinants of the Fab fragment, block antigen binding by purified idiotype-bearing murine antibodies, and provide useful probes for visualization and analysis of immunoglobulin-like surface receptors of rodent B- and T-cells.


Immune responses to some proteins and synthetic polypeptides in inbred strains of rats. The antibody responses to DNP-bovine gamma globulin, bovine gamma globulin, human serum albumin, ovalbumin, pepsin and five synthetic polypeptides were examined in strains of inbred rats representative of eight common major histocompatibility complex (RTI) haplotypes. With each antigen the antibody response varied considerably among strains, and the data provide many potential strain and antigen combinations with which to study the genetic control of the immune response.


Properties of an Fc gamma-binding protein isolated from human leukemic B cells. A selectively Fc gamma-binding protein was isolated from purified and radioiodinated cell membranes from two cases of B-type chronic lymphocytic leukemia and one case of B-type prolymphocytic leukemia by binding to IgG aggregates, horseradish peroxidase-anti-peroxidase IgG complexes, and sheep erythrocyte membrane sheets densely coated with IgG. This protein could not be isolated from the cell membranes of an Fc gamma-receptor-negative chronic lymphocytic leukemia of the T type or from membranes of human erythrocytes. The Fc gamma-binding protein was efficiently solubilized by a mixture of Na-EDTA and 2-mercaptoethanol, but not with one of these agents alone, indicating that both divalent cations and disulfide bridges are involved in the linkage of the Fc gamma-binding protein to the cell membrane. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the Fc gamma-binding protein revealed an apparent mol wt of 28,000 and in isoelectric focusing it showed an isoelectric point of 5.5. The electrophoretic mobility of the 28,000-dalton protein did not change after reduction and alkylation. It was determined that the NH2-terminal amino acid of the protein was glycine. The isolated protein was unable to agglutinate antibody-coated erythrocytes. These findings suggest that the 28,000-dalton IgG-affined protein was composed to O2-enriched buffer lacking reducing agents, the 28,000-dalton protein aggregated to a 115,000-dalton molecule. The isolated Fc gamma-binding protein proved to be different from C1q or its subunits.


Binding of purified, soluble major histocompatibility complex polypeptide chains onto isolated T-cell receptors. I. Reactivity against allo- and self-determinants. In this study, we tried to get information about the fine antigen-binding ability of purified, soluble, idiotype-positive T-cell receptor molecules. Lewis anti-DA T-cell receptors were purified from normal Lewis serum by the use of anti-idiotypic immunosorbent and sodium dodecyl sulfate-polyacrylamide gel, and were coupled to cyanogen bromide-activated Sepharose 4B. In parallel, Lewis anti-DA, Lewis anti-BN, and DA anti-Lewis alloantibody immunosorbents were prepared. The major Ag-B chain (44,000 daltons) and the two polypeptide chains (34,000 and 27,000 daltons) of Ia were purified from Lewis, DA, and BN lymphocytes and absorbent on the above-mentioned immunosorbents. We found that the major Ag-B chain as well as the two Ia chains were bound to the alloantibody columns if they were derived from the corresponding allogeneic strain. No retaining ability for self-major histocompatibility complex (MHC) or third-party MHC chains was noted with the alloantibody immunosorbents. When using immunosorbents made up of idiotypic T-cell receptors, only two MHC polypeptides of the relevant allo-MHC type were retained, namely, the Ag-B and the heavy Ia chains. No detectable activity was observed when testing the same column for reactivity against third-party MHC polypeptide chains. However, the Lewis anti-DA T-cell receptors could be shown to display weak, but significant, reactivity toward one Lewis MHC polypeptide chain, that is, the heavy chain of Ia type.


Regulatory mechanisms in cell-mediated immune responses. VIII. Differential expression of I-region determinants by suppressor cells and their targets in suppression of mixed leukocyte reactions. The phenotypic expression of I-region determinants on cells producing and responding to MLR suppressor factor (MLR-TsF) was established in these studies. Alloantigen-activated MLR suppressor T cells (MLR-Ts), which produce MLR-TsF bearing gene products of the I-C subregion, were exposed to anti-I subregion sera and complement (C) before in vitro culture for MLR-TsF production. Suppressor activity was prevented by removal of cells bearing I-C determinants, whereas elimination of cells expressing I-A/B determinants had no effect. Interestingly, cytotoxic elimination of cells displaying I-J determinants also prevented MLR-TsF production. Admixture of anti-I-J and I-C antiserum-treated cells for MLR-TsF production failed to reconstitute suppressor activity, indicating that I-C and I-J gene products are expressed on a single population of cells critical to MLR suppression, rather than on distinct interacting subpopulations. Anti-I-C serum activity specific for I-C+ MLR-Ts was removed by adsorption with nylon wool-nonadherent splenic T cells and concanavalin A-activated thymocytes; adsorption with splenic B cells from anti-Thy-1,2 serum and C-treated spleen failed to remove relevant anti-I-C activity. These data suggest that regulatory I-C molecules, like I-J molecules, are preferentially expressed on T lymphocytes. Expression of I-C, or other I-region molecules on responder cell targets of MLR-TsF activity was also investigated. Responder cells were pretreated with anti-I subregion-specific sera in blocking or complement-dependent cytotoxic protocols before addition to MLR with MLR-TsF. Neither blocking nor the cytotoxic removal of cells bearing I-C or other I-region determinants from MLR responder populations interfered with MLR-TsF suppression. Because it has previously been demonstrated that MLR-TsF interacts optimally with activated, I-C syngeneic target cells, blocking and cytotoxic studies with anti-I subregion sera were also performed with responder cells activated by 24 h culture in MLR in the absence of MLR-TsF. Brief MLR-TsF pulse after antiserum treatment generated marked suppression regardless of blocking or absence of cells bearing serologically detected I-region determinants. I-C restricted suppression may thus be mediated not by interaction with I-C-bearing cells, but by target cells which exist in requisite association with populations of I-C+ cells.


Viral specificity of H-2-restricted T killer cells directed against syngeneic tumors induced by Gross, Friend, or Rauscher leukemia virus. Cytolytic T lymphocytes (CTL) were generated against murine tumors induced by Gross, Friend, or Rauscher leukemia virus (LV) in syngeneic mixed leukocyte-tumor cell cultures. Analogous to the patterns of specificity observed with antibodies to LV-induced cell surface antigens, CTL could be classified into two major groups of specificity. Tumor cells induced by Friend, Moloney, or Rauscher virus and positive for the FMR antigen were killed by syngeneic CTL immune to any one of these three LV; the same CTL, however, were incapable of killing syngeneic tumor cells induced by Gross LV. The converse was true for Gross LV-specific CTL: these CTL were specific for syngeneic tumor cells expressing the Gross virus-associated cell-surface antigen (GCSA), and not the FMR antigen. The H-2 specificities of the two groups of LV-immune CTL were also compared, because in both cases, CTL were restricted in their killing activity to H-2-identical tumor target cells. When CTL from single strains of mice were generated against syngeneic FMR- or GCSA-positive tumor cells, differences were observed with respect both to the requirement for the expression of compatible H-2K or H-2D specificities, and to the intensity of the CTL response in congenic mice of the H-2b, H-2d, and H-2k haplotypes.


Antigen- and receptor-driven regulatory mechanisms. I. Induction of suppressor T cells with anti-idiotypic antibodies. Delayed-type hypersensitivity (DTH) to the azobenzenearsonate (ABA) hapten can be readily induced in A/J mice injecting ABA-coupled syngeneic spleen cells subcutaneously. To further characterize this T-cell-dependent immunological phenomenon, the effect of passively administered anti-cross-reactive idiotype common to anti-ABA antibodies of A/J mice (CRI) antibodies on the development of ABA-specific DTH was investigated. Animals given daily injections (of minute amounts) of anti-CRI antibodies subsequent to immunization with ABA-coupled cells show significant reduction of ABA specific responses. This inhibition is antigen specific and requires the intact immunoglobulin molecule, as F(ab')2 treatments were ineffective in suppressing the reaction. Investigations of the mechanism of the anti-CRI-induced suppression of ABA DTH revealed that the observed suppression is a result of the activation of suppressor cells. Spleen cells taken from animals which received anti-CRI antibodies were able to adoptively transfer suppression to naive recipients. This suppression was shown to be mediated by T cells, as anti-Thy1.2 plus complement completely abrogated the transfer of suppression. In addition, animals pretreated with low doses of cyclophosphamide were not suppressed by the administration of anti-CRI antibodies. The genetic restriction of anti-CRI-induced suppression was demonstrated. Antibodies to the major cross-reactive idiotype, (CRI) associated with anti-ABA antibodies in A/J mice were unable to suppress the development of DTH to ABA in BALB/c mice (H-2d, Igh-1a). Such antibodies were, however, fully active in suppressing ABA DTH in the allotype-congenic C.AL-20 strain which has an allotype (Igh-1d) similar to that of A/J (Igh-1e) on a BALB/c background, and which produces humoral antibodies with the CRI.


Antigen- and receptor-driven regulatory mechanisms. II. Induction of suppressor T cells with idiotype-coupled syngeneic spleen cells. Anti-p-azobenzenearsonate (ABA) antibodies, coupled covalently to normal syngeneic spleen cells and then given intravenously to normal animals, were found to be potent tolerogens for delayed-type hypersensitivity (DTH) to ABA. The ability of the antibody-coupled cells to induce tolerance was determined to be a result of the cross-reactive idiotype (CRI+) fraction of the antibodies, because anti-ABA antibodies lacking the CRI+ components when coupled to spleen cells were unable to cause any significant inhibition. Furthermore, genetic analysis revealed that the ability of CRI-coupled cells to inhibit ABA-specific DTH is linked to Igh-1 heavy chain allotype, in as much animals which possess heavy chain allotypes similar to that of A/J were sensitive to this inhibition. Adoptive transfer experiments provided evidence that CRI-coupled cells induce suppressor cells, and spleen cells or thymocytes from animals received CRI-coupled cells were able to transfer suppression to naive recipients. In addition, treatment with anti-Thy1.2 serum plus complement completely abrogated their ability to transfer suppression. Thus, this active suppression is a T-cell-dependent phenomenon. In investigating the specificity of these suppressor T cells, it was found that they functioned in an antigen-specific manner and were unable to suppress the development of DTH to an unrelated hapten 2,4-dinitro-1-fluorobenzene.


Antigenicity of the infected-erythrocyte and merozoite surfaces in Falciparum malaria. The antigenicity of altered structures induced by Plasmodium falciparum in the membranes of infected Aotus monkey and human erythrocytes was examined. Antisera were obtained from monkeys made immune to malaria. Bound antibodies were shown to be localized on the knob protrusions of infected erythrocytes of both human and monkey origin and from both in vitro and in vivo infections. Therefore, P. falciparum infection has produced similar antigenic changes in the erythrocyte surfaces of both man and monkey. Uninfected erythrocytes and all knobless-infected erythrocytes bound no antibody from immune sera. Strains of P. falciparum from widely different geographic areas that were cultured in vitro in human erythrocytes induced structures (knobs) which have common antigenicity. Merozoites were agglutinated by cross-linking of their cell coats when incubated with immune sera. The binding of ferritin-labeled antibody was heavy on the coats of both homologous and heterologous strains of the parasite, indicating that the merozoite surfaces of these strains share common antigens.


Presence of plasma cells binding autologous antibody during an immune response. Spleen and other lymphoid tissues of rabbits immunized with human serum albumin (HSA) and human lactoferrin (LF) were examined for the presence of cells forming anti-idiotype antibodies. To detect these cells, IgG, F(ab')2, or Fab' of specific antibodies were isolated, fluorochrome-tagged with tetramethylrhodamine isothiocyanate, and used as an idiotypic marker to detect splenic plasma cells that are producing anti-idiotypic antibody. By this procedure, we were able to demonstrate anti-idiotypic cells in surprisingly high numbers. For example, in six rabbits immunized with HSA for periods ranging from 36 to 542 d, the percentage of Ig-positive cells that stained with autologous idiotype ranged from 0.7 to 44; furthermore, cross-reactivity was observed among seven different anti-HSA preparations and two anti-LF antisera. The isotype of anti-idiotypic cells, determined by costaining with fluorescein isothiocyanate-labeled goat Fc-specific anti-rabbit Ig, was shown to be predominantly IgG. These findings provide evidence of the presence of plasma cells producing antibody to autologous idiotype during a vigorous immune response.


More than one antigen contributes to the immunogenicity of Neisseria gonorrhoeae in the guinea pig chamber model. Gonococci adapted to growth in guinea pig chambers [strain BS4 (agar)] were predominantly smooth organisms and produced a type-specific antigen. A vaccine prepared by treating these gonococci with formalin, protected guinea pig chambers against homologous challenge in contrast to a similarly treated laboratory strain (BSDH) which had been selected in vitro from the same parent strain and which did not produce the type-specific antigen. Surface washes of BS4 (agar) contained the type-specific antigen but attempts to immunize guinea pigs with complexes of rabbit antibody with this antigen excised from gels failed. However, good immunity could be produced by combining such complexes with formalin-killed rough gonococci (strain BS4R), lacking the type-specific antigen, which were found in some chambers of challenged guinea pigs that had been immunized with the complexes. Hence, at least two antigens -- one the type-specific antigen and the other(s) possessed by both BS4 (agar) and BS4R -- are needed for immunogenicity. Surface washes of BS4 (agar) and BS4R contained three antigens, distinct from the type-specific antigen, which might complement it in producing immunity. Similar antigens were present in surface washes of five fresh isolates from human urethral pus, but only a few organisms from these isolates possessed antigens similar to the type-specific antigen. The variability of gonococci in antigenicity, immunogenicity and probably virulence is discussed.


Further studies of the antigenic properties of H3N2 strains of influenza A isolated from swine in South East Asia. H3N2 strains of influenza A isolated from swine in Hong Kong were compared with human strains of H3N2 influenza A variants in reciprocal HI tests using ferret sera. One isolate from swine was indistinguishable from A/Hong Kong/68, one set of viruses isolated in 1976 and 1977 was most related to A/Hong Kong/68 but was not identical to it, two isolates from 1976 were 'bridging strains' that cross-reacted equally with the contemporary variants A/Victoria/3/75 and A/Texas/1/77, similarly to a small number of recent human isolates, and two isolates from 1977 were similar to A/Victoria/3/75. These general relationships were supported by neuraminidase inhibition tests. The findings confirm and extend previous results indicating that swine may be a reservoir of old and novel variants of influenza A H3N2 strains related to those that infect man.


Malignant fibrous histocytoma producing spinal cord compression. Three cases of malignant fibrous histiocytoma presented as primary paraspinal tumours causing extradural spinal cord compression. Study of these cases suggests that pain relief is considerable after laminectomy but pain may recur with further tumour growth. Total removal is unrealistic and diagnosis, difficult. The prognosis in terms of continuing neurological deficit after surgery followed by local radiotherapy appears poor.


Adult type mucolipidosis with beta-galactosidase and sialidase deficiency. Histological and biochemical studies. A case of adult type mucolipidosis with beta-galactosidase and sialidase deficiency is described. This patient, a woman aged 20, had mental retardation, macular cherry-red spots, corneal clouding, gargoyle-like face, cerebellar ataxia, myoclonus and convulsions beginning at the age of 14. Bony deformities, vacuoles in the peripheral lymphocyte and foamy cells in the bone marrow were also noted. Biopsy study of the sural nerve and vermiform appendix disclosed many vacuoles in almost every kind of cells, although the accumulated substance in these vacuoles could not be characterized histochemically or ultrastructurally. Deficient leukocyte beta-galactosidase and sialidase were confirmed. There was increased urinary sialoglycopeptide and increased siliac acid and hexosamine in the glycoprotein of lymphocytes. Leukocytes sialidase activites of the parents were 30 to 50% of the control values. These results suggest a genetic defect of sialidase.


Influence of temperature on isometric contraction and passive muscular tension in paramyotonia congenita (Eulenburg). Four patients without symptoms of episodic hyperkalemic weakness from two families with paramyotonia congenita (Eulenburg) are described. 1. Maximum voluntary muscle contraction of the upper and lower arm was studied under isometric conditions at different temperatures. If the temperature was lowered stepwise, distinct paresis occured at 32--31 degrees C which increased with the amount of muscular effort. The upper arm muscles, however, developed weakness gradually after cooling. 2. During cooling of the resting muscle, the EMG showed dense spontaneous activity of the fibrillary type, which decreased again at about 30 degrees C. It can be assumed that in paramyotonia congenita cooling produces muscle cell membrane depolarization which at a critical level causes the firing of action potentials and finally muscular paresis. 3. Increasing muscular stiffness can be interpreted as abnormally slow muscular relaxation after isometric contraction. In the forearm muscles the time to 3/4 relaxation after cooling was about six times normal, in the upper arm muscles only two times normal. As an additional parameter the mechanical resistance to passive stretching of a muscle has been studied. This passive muscular tension increased simultaneously with the onset of weakness. 4. The close relation between weakness and stiffness suggest that both symptoms are caused by the same basic defect which is probably located in the sarcolemma. It is suggested that a defect of the sodium channel causes a cooling-dependent increase in sodium conductance. Raised intracellular sodium causes in the first place membrane depolarization, and in the second place depression of calcium reuptake through competition by sodium for calcium binding sites. This would explain muscle stiffness and delayed relaxation as well.


Partial Klüver-Bucy Syndrome following probably herpes simplex encephalitis. An autopsy case of probable herpes simplex encephalitis (HSE) showed the partial Klüver-Bucy syndrome from 1 to 7 months after the onset, which consists of strong oral tendencies, emotional changes and possible hypersexuality. Several cases of the syndrome in definite or probable HSE were reviewed in this paper. In these cases including our case, the partial or complete Klüver-Bucy syndrome has appeared from some interval (2 weeks--1 month) after the acute phase, so that the syndrome should be noticed in survived or prolonged cases of HSE, as significant clinical symptoms suggesting bilateral involvement of the temporal lobes and limbic areas.


Inhibition of reverse transcriptase activity by benzophenanthridine alkaloids. Benzophenanthridine alkaloids, fagaronine 4, O-methylfagaronine 5, nitidine 1, allonitidine 3 and methoxydihydronitidine 2 have been shown to posses inhibitory activity against reverse transcriptase of RNA tumor viruses. The enzyme inhibition (50%) by these alkaloids was found in the range of 6-60 microgram per milliliter of the reaction mixture when polynucleotide-oligodeoxynucleotide complexes were used as template primers. The results suggested that the benzophenanthridine alkaloids interacted with the template primers (particularly of the A:T base pairs) and not with the enzyme proteins. Kinetics reaction of the reverse transciptase inhibition showed that the alkaloids stopped the DNA polymerase synthesis instantly, probably by interacting with the template primer.


An unusual form of measles meningoencephalitis. A report of two cases. Two patients are reported with a chronic progressive illness characterized by dementia, ataxia and spasticity. There were no myoclonic jerks and both had normal electroencephalograms (EEG). Pathological findings in three brain biopsies were those of viral meningoencephalitis with perivenous demyelination. Serological data in both patients indicated the presence of measles virus infection. Intracytoplasmic structures resembling measles virus nucleocapsids were found in the brain biopsy of one patient. Immunofluorescent staining showed antibody in the temporal lobe biopsy of both patients. It is suggested that these patients are examples of a chronic form of measles meningoencephalitis hitherto undescribed.


Chronic cluster headache: response to lithium treatment. A clinical trial of lithium carbonate was undertaken in a group of 20 patients suffering from chronic cluster headache. Blood levels of lithium were determined at regular intervals to monitor the therapeutic dosage. Lithium was found to be an effective prophylactic agent for chronic cluster headache patients. The effectiveness of lithium was evident in less than a week after the beginning of treatment. It is not clear in what way lithium may act on cluster headache.


A case of glioblastoma with multiple centers above and below the tentorium. A case of multicentric glioblastoma with the clinical and histopathological findings is presented in which three lesions are located above and below the tnetorium. Multicentric glioblastomas are those which have no macroscopic or microscopic connection.


Mitochondrial abnormalities of late motor neuron degeneration following poliomyelitis and other neurogenic muscular atrophies. A case of late motor neuron degeneration following poliomyelitis with abnormal mitochondria in muscle fibers is presented with two additional cases of systemic neurogenic muscular atrophy (Charcot-Marie-Tooth disease). Muscle biopsy revealed a neurogenic pattern of variable severity in all cases. Subsarcolemmal zones of hyperactivity and hyperpositive intermyofibrillar collections of granular material present in a variable proportion of type I fibers were demonstrated by oxidative enzymes. Ultrastructurally they corresponded to abnormal mitochondria, with paracrystalline inclusions in one case. The finding is discussed in the light of the previous literature on mitochondrial myopathies. Mitochondrial alterations are not specific and their significance in neurogenic conditions is debated.


Subarachnoide hemorrhage due to S. mansoni. A rare etiology. A case of schistosomiasis is described in a native of Rio de Janeiro, where, apart from infection of the intestines, a meningitic granulomatous reaction was provoked around eggs. Intracerebral and subsequently subarachnoid hemorrhage occurred. The mode of infection is disucssed.


Herniation of an intervertebral disc with cauda compression syndrome in childhood. A case of a lumbosacral disc herniation of traumatic origin with compression of the cauda equina in a child is reported. The problems of early diagnosis, timely operative procedure, and prognosis of neurological deficit of operated disc herniation in childhood are discussed.


Discovery of an epidermoid of the skull concomitant with a homolateral subacute traumatic subdural hematoma. The case of man of 32 years with an epidermoid of the left side of the skull is reported. The tumor itself was asymptomatic and was discovered accidentally because of a subdural hematoma on the same side. After a closed head injury, this patient had symptoms of an intracranial space occupying lesion (hematoma) on the left after a symptom-free interval. The left carotid angiogram demonstrated the characteristic picture of a subdural hematoma in the left parieto-temporo-occipital region. At the same time, characteristic changes in the skull on the same side, which were more apparent after removal of the hematoma, suggested an epidermal tumour of the skull. This was verified at operation.


Sensitivity of the short-range spinal interneurons of the cat to experimental spinal cord trauma. The technique of retrograde axoplasmic transport was used to demonstrate the effect of experimental spinal cord injury on the spinal interneurons in the upper lumbar and lower thoracic segments of cats. Force of varied intensity was applied to the dorsal surface of the spinal cord and horseradish peroxidase (HRP) was injected into the next caudal segment. A large impact (250 to 350 gm-cm) inducing permanent paraplegia of the hind legs blocked the axoplasmic transport instantaneously in both cranial and caudal directions. If 1 week elapsed between the trauma and injection, neurons cranial to the trauma did not show any evidence for retrograde axoplasmic transport, while few neurons in the caudal direction were labeled with HRP. A moderate impact (150 gm-cm) which rendered the animals only transiently paraplegic spared the axoplasmic transport in some neurons both cranially and caudally to the injection. No obvious recovery or additional loss in the number of HRP-labeled neurons could be found in the cats if the injections followed the trauma by 1 week. The loss of spinal cord neurons following the injury seems to be the immediate mechanical consequence of the trauma.


Development of annular gap junctions in guinea pig epithelia. Epithelia from skin, palate, buccal mucosa and dorsal tongue were studied in normal and zinc deficient guinea pigs by transmission electron microscopy. Gap junctions (nexuses) were observed primarily in the basal and prickle cell layers. They were also seen to form bulging and looping configurations which appeared to pinch off into the cytoplasm forming what are recognized as annular gap junctions. These may be attached to the surface or represent separate, intracytoplasmic, nexus-bound inclusions. Their contents varied, but most often they contained glycogen and ribosomal-like particles. Various cell organelles were also seen entrapped, most common of which were membrane coating granules. The development of these annular gap junctions is a prominent feature of many epithelia. It has been suggested that they probably serve to maintain a synchronized differentiation throughout the various epithelial layers.


Effect of ricinoleic acid and other laxatives on net water flux and prostaglandin E release by the rat colon. Ricinoleic acid, oleic acid, dioctyl sodium sulphosuccinate, deoxycholic acid, sennoside A + B and mannitol reduced or reversed water flux from lumen to blood in rat colon in situ. Stearinic acid was without any effect. Ricinoleic acid, oleic acid, dioctyl sodium sulphosuccinate, deoxycholic acid and sennoside A + B stimulated release of PGE-like material into the colonic lumen whereas the osmotic laxative mannitol and stearinic acid did not. Inhibition of PGE biosynthesis by pretreatment of the rats with indomethacin significantly reduced (but did not abolish) the effect of ricinoleic, oleic and deoxycholic acids on net water flux and PGE release. Indomethacin reduced the effect of dioctyl sodium sulphosuccinate and of sennoside A + B on PGE release but not their effect on the net water flux. The effect of mannitol was not influenced by indomethacin. The amount of PGE release in experiments with ricinoleic acid, oleic acid, stearinic acid and dioctyl sodium sulphosuccinate (with and without indomethacin) showed a good correlation (r = 0.99) with the change in net water flux. Deoxycholic acid, sennoside A + B and mannitol did not show this correlation. It is assumed that the action of non-osmotic laxatives is partially mediated by PGE, although other mechanisms also seem to be involved in their mode of action.


Ion-pair liquid chromatographic assay of decongestants and antihistamines. The quantitative determinations of combinations of antihistamine and decongestant drugs including phenylephrine, dl-ephedrine, psi-ephedrine, phenylpropanolamine, pyrilamine, pheniramine, l-ephedrine, chlorpheniramine, brompheniramine, oxymetazoline, naphazoline, and antazoline contained in solid and liquid dosage forms are described. All active ingredients except the ephedrine optical isomers were separated from other ingredients with ion-paired high-pressure liquid chromatography. Manipulation of the mobile phase either by changing the hydroalcoholic ratio or by changing the alkyl chain length of the counterion (sulfonic acid) for achieving optimum separations is discussed. The method is simple, short, accurate, and precise.


Two avian sarcoma virus mutants with defects in the DNA polymerase-RNase H complex. Two mutants of avian sarcoma virus which exhibit different phenotypes have been analyzed for the properties of their RNA-dependent DNA polymerase and RNase H activities. LA 338 is a complex multiple mutant with at least one lesioneach in transformation and replication functions. The purified RNA-dependent DNA polymerase-RNase H complex from the mutant is twofold more thermolabile than that from the wild-type parent. A peculiarity of this mutant is that the ability of the enzyme to respond to synthetic template-primers is lost more rapidly than is the response to native RNA as template. The mutant enzyme cannot be protected from inactivation by the addition of synthetic template-primers. LA 672 represents a different phenotype among reverse transciptase mutant, showing a "late"-acting block in replication which affects only production of progeny by infected cells grown at the nonpermissive temperature. The purified DNA polymerase-RNase H complex of LA 672 is not thermolabile; rather, progeny grown at the nonpermissive temperature yield purified enzyme with a 20-fold-reduced specific activity in both DNA polymerase and RNase H. The content of reverse transcriptase protein in such noninfectious progeny, furthermore, did not appear to be significantly diminished since immunologically active enzyme could be demonstrated in a competition test for anti-reverse transcriptase antibody and since beta and alpha subunits of reverse transcriptase could be identified after polyacrylamide gel electrophoresis of partially purified enzyme preparations. The amounts of beta and alpha from the mutant were about twofold lower.


Deletion mutant of the Bratislava-77 strain of Rous sarcoma virus containing a fusion of the group-specific antigen and envelope genes. The genetic compositions of two independently derived preparations of the Bratislava-77 strain (B77) of Rous sarcoma virus were analyzed after each was passaged seven or more times in duck embryo fibroblasts. RNase, T1-resistant oligonucleotide fingerprint analysis of virion RNA from both preparations of duck-passaged B77 revealed the presence of two large noncontiguous deletions. Approximately 75% of the RNAs contained a deletion which spans oligonucleotides 304 to 4 on the viral genome (about 3,500 nucleotides) and encompasses all of the B77 polymerase gene. More than 90% of the RNAs also contained a deletion which spans src-specific oligonucleotides 6 and 5(about 2,200 nucleotides) and is identical to the deletion observed in transformation-defective B77. Virion RNA from duck-passaged B77 also contained two oligonucleotides (D1 and D2) not observed in the RNA of B77 virus grown on chicken embryo fibroblasts. Analysis of the virion RNA of duck-passaged B77 by denaturing agarose gel electrophoresis revealed four major subunits with molecular weights of 3.40 x 10(6), 2.65 x 10(6), 2.25 x 10(6), and 1.55 x 10(6). Whereas the 3.40- and 2.65-megadalton (Mdal) RNA species comigrated with the nondefective and transformation-defective RNAs of B77 propagated on chicken embryo fibroblasts, no counterparts to the 2.25- and 1.55-Mdal RNAs were observed in the RNA of B77 grown on chicken embryo fibroblasts. Oligonucleotide fingerprint analysis of these RNA species revealed that the 2.65-Mdal RNA contains the src-specific deletion and that 2.25-Mdal RNA contains the polymerase region deletion; both of these deletions were observed in the 1.55-Mdal RNA, which was the major RNA subunit species detected in duck-passaged B77. The new oligonucleotides (D1 and D2) observed in the duck-passaged virus were present in the 2.25- and 1.55-Mdal RNA species in vitro and in vivo and directs the synthesis of a 130,000-dalton protein (p130). p130 contains antigenic determinants specific for p27 (gag gene) and gp85 (env gene) but does not contain sequences which cross-react with antisera directed against the alpha beta form of RNA-dependent DNA polymerase (pol gene). This RNA, therefore, is generated by a fusion of the gag and env genes of Rous sarcoma virus B77.


Production of monoclonal antibodies against nucleocapsid proteins of herpes simplex virus types 1 and 2. We prepared mouse hybrid cell lines which produced antibodies against herpes simplex virus type 1 and 2 nucleocapsids. Cell lines 1D4 and 3E1, respectively, secreted immunoglobulin G1 herpes simplex virus type 1 and immunoglobulin G1 herpes simplex virus type 2 antibodies which immunoprecipitated proteins designated p40 and p45 from homologous nucleocapsid preparations but precipitated no proteins from heterologous preparations. In contrast, guinea pig antisera prepared against either herpes simplex virus type 1 or 2 p40 precipitated p40 and p45 from both homologous and heterologous preparations. These findings suggest that p40 and p45 possess similar antigenic determinants and that the monoclonal antibodies that were tested reacted preferentially with the homologous determinants.


The ultrastructural changes of prostate adenocarcinoma following external beam radiation therapy. Ultrastructural studies were done to determine the effects of radiation therapy on prostatic adenocarcinoma. Twenty patients were included in the study: 6 with benign hyperplasia, 4 with untreated adenocarcinoma and 10 with adenocarcinoma who had received radiation therapy. Benign and malignant ultrastructural characteristics were established. On the basis of these characteristics 6 of the 10 radiated prostates were believed to have tumor cells present after therapy. Of the remaining 4 patients 3 had ultrastructural changes suggesting radiation effects and a possibly altered malignant potential.


Human lung tumor-associated antigens of 32,000 daltons molecular weight. Lung tumor-associated antigens of approximately 32,000 daltons were recognized by the use of sensitive radioimmunoassays and rabbit antisera, one raised against an extract of pooled human malignant lung tissues and another raised against a cell line derived from a human squamous cell carcinoma of the lung. These antigens differ from antigens described previously, including carcinoembryonic antigen and alpha-fetoprotein. The antigens were detected on 13 of 13 lung tumors (of all histologic types), fetal tissue, normal brain, 2 of 8 colon tumors, 2 of 9 prostate tumors, and 2 of 3 breast tumors, as well as on cell lines derived from lung tumors, neuroblastoma, human amnion, colon adenocarcinoma, and bladder tumors. They were not detectable on normal lung, liver, kidney, colon, or prostate tissues or on cell lines derived from osteosarcoma, fetal lung fibroblasts, transitional cell carcinoma, and squamous cell carcinoma of the skin. Lung tumors of different histologic types were concluded to express common, tumor-associated oncofetal antigens that are found less often on tumors of other organs.


Evaluation of a "nude" mouse-human tumor panel as a predictive secondary screen for cancer chemotherapeutic agents. Nine established human melanoma tissue-cultured cell lines heterotransplanted in C57BL/6 "nude" mice were exposed to each of 4 chemotherapeutic agents of known clinical activity against human melanoma. Two of the therapeutic agents, 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and 5-(3,3-dimethyl-1-triazino) imidazole-4-carboxamide (DTIC), are known to be active against human melanoma; the other two, adriamycin and 5-azacytidine, are known to be inactive. Sterile saline served as a control agent. In 2 cell line heterotransplants, the control tumor spontaneously regressed. Of the 7 cell lines that remained for evaluation, 4 were sensitive to DTIC, 1 was sensitive to BCNU, and none was sensitive to adriamycin or 5-azacytidine. These data indicate that the nude mouse-human tumor model may be a predictive secondary screen for cancer chemotherapeutic agents.


[Clinical study of the antiarrhythmic action of Mexityl in ventricular disorders of the cardiac rhythm]. The anti-arrhythmic activity of mexitil was studied in 36 patients with frequent, prognostically unfavourable ventricular extrasystoles of various etiology. Quantitative and qualitative assessment of rhythm disorders was accomplished by bicycle ergometry and ECG recording for 24 hours by means of portable monitors. The disorders of rhythm were appraised prior to treatment, during mexitil medication in daily doses of 750--1200 mg/24 hrs administered for 3--5 days, and against the background of placebo. The effect of a single 250 to 600 mg dose of mexitil was appraised by monitoring the ECG for three and a half hours. The concentration of mexitil in blood plasma and saliva was tested by gas-fluid chromatography. A positive effect of mexitil treatment manifested in complete correction of ventricular extrasystoles or the reduction of their number by more than half and complete elimination of group extrasystoles and paroxysms of ventricular tachycardia was produced in 66 per cent of patients according to the daily ECG monitoring finds and in 76 per cent of patients according to the results of bicycle ergometry. The antiarrhythmic effect developed 62 minutes, on the average, after oral administration of a single 250--600 mg dose of mexitil and lasted 5--8 hours and more. The therapeutic concentration of mexitil ranged from 0.6 to 0.9 micrograms/ml in blood plasma and from 10.3 to 15.8 micrograms/ml in saliva. In 36 per cent of patients side effects (mainly muscular tremor, lassitude and headache) were noted. It is concluded that mexitil possesses high anti-arrhythmic activity in ventricular extrasystole. To avoid side effects it is recommended to begin treatment in a dose of 250 mg given three times daily, and if necessary gradually increase the single dose to 400 mg and the daily dose to 1200 mg.


[Current status in the treatment of breast cancer. II. Adjuvant chemotherapy, palliative polychemotherapy, chemoimmunotherapy--rating and results (author's transl)]. There is no well defined group of patients with primary breast cancer which benefits from combination chemotherapy as an adjuvant treatment, since, at present, the effect of this therapy in respect to the duration of disease-free interval, survival, and possible long-term side effects remain unknown. Therefore, controlled studies need to be initiated. Similarly, there seems to be no beneficial effect from unspecific immunotherapy. As far as combination chemotherapy in advanced breast cancer is concerned, we review on four different protocols which proved to be quite successful in our hands: adriamycine/cyclophosphamide (AC), cyclophosphamide/methotrexate/5-fluorouracil (CMF), CMF/vincristine/prednisone (CMFVP), and adriamycine/vincristine plus CMF plus Tamoxifen.


Controlled trials of a polyvalent pseudomonas vaccine in burns. A polyvalent pseudomonas vaccine has been tested in controlled clinical trials at two burns units, in Birmingham and New Delhi, in children and adults with burns more than 15% full skin thickness. None of the vaccinated patients in either trial showed blood cultures containing Pseudomonas aeruginosa, and vaccinees showed raised titres of protective antibody and increased phagocytic activity against Ps. aeruginosa. In the New Delhi unit, where death from Ps. aeruginosa infection is common, the mortality in adults was reduced from 40.6% (13/32) in the unvaccinated group to 6.6% (2/30) in the vaccinated group, and in children from 20.8% (5/24) in the unvaccinated group to 4.8% (1/21) in the vaccinated group.


Early determination of androgen-responsiveness is important in the management of microphallus. Of fourteen boys with microphallus who were studied consecutively, nine had hypopituitarism that might otherwise have not been recognised. All fourteen were treated with low-dose, intramuscular testosterone enanthate administered in one or two 3-month courses. In each case, penile size was increased into the normal range without evoking either a significant increase in height velocity or an advancement of skeletal maturation. Before considering surgical sex reversal in infants and young children with microphallus, it is essential to assess whether treatment with intramuscular testosterone will induce clinically significant growth of the microphallus.


Incidence and significance of antibodies to delta antigen in hepatitis B virus infection. A microtitre solid-phase blocking radioimmunoassay (RIA) for antibody to the hepatitis B virus (HBV)-associated delta antigen was specific and detected anti-delta antibody at dilutions of serum of up to 10(6). Analysis of sera from HBsAg-negative subjects and different categories of HBsAg carriers from different regions confirmed the association of anti-delta antibody with HBV infection. Anti-delta antibody was detected in persistently high titres in 19.1% and 2.6% of sera from patients with chronic hepatitis and symptomatic chronic carriers, respectively, and was not detected in the sera of HBsAg-negative controls. Anti-delta antibody appeared transiently and in low titres (less than 1:500) in 4.8% of sera from patients with acute type B hepatitis. The presence and persistence of anti-delta antibody seem to be associated with chronic HBV infection and the development of progressive liver damage.


Prenatal diagnosis of Farber's disease.

Prenatal diagnosis of Farber's disease. Two pregnancies at risk for Farber's disease were monitored with amiocentesis at 15 and 16 weeks' gestation. In the first pregnancy tested, cultured amniotic-cell ceramidase activity was 7.8% of the control mean and an affected fetus was predicted. The pregnancy was terminated at 22 weeks' gestation and the diagnosis was confirmed by the demonstration of considerably elevated renal and hepatic ceramide concentrations and severe reduction of ceramidase activity in fetal brain and cultured fibroblasts. In the second pregnancy tested, cultured amniotic-cell ceramidase activity was within the control range, and the prediction of an unaffected fetus was confirmed in the newborn.


Effects of 1-desamo-8-D-arginine vasopressin on behaviour and cognition in primary affective disorder. DDAVP (1-desamino-8-d-arginine vasopressin), a synthetic analogue of vasopressin with prolonged half-life and high antidiuretic and low pressor activity, was given in a double-blind placebo-controlled trial to four patients with major affective illness. Three of four patients showed highly significant and consistent improvements in tests designed to measure the formation, encoding, and organisation of long-term trace events in memory. Two patients also showed a significant but less consistent amelioration of other depressive symptoms during DDAVP treatment. These findings implicate central vasopressin function in the processing of information and possibly other aspects of affective illness.


Importance of the neural predilection of Mycobacterium leprae in leprosy. It is suggested that continuous leakage of bacilli into the circulation from a primary focus of intraneural infection may simultaneously initiate bacillary dissemination and the suppression of cell-mediated immunity. Both these features are essential for the development of lepromatous leprosy. Nerve involvement in leprosy, previously thought of as a diagnostic feature of the disease and as a complication of therapy, may represent an essential phase in the cycle of infection and reinfection by Mycobacterium leprae.


Cyclosporin A initially as the only immunosuppressant in 34 recipients of cadaveric organs: 32 kidneys, 2 pancreases, and 2 livers. 34 patients treated with cyclosporin A received 36 cadaveric organ allografts (32 kidneys, 2 pancreases, and 2 livers), 26 kidneys are still supporting life, 3 after more than a year; the pancreases and livers are also functioning. 20 patients are not receiving steroids, and 15 of these have not had any additional immunosuppressive agents. In these patients infectious complications have not been severe, but a gastroduodenal lymphoma has developed in 1 patient. 6 patients were given 'Cytimum' (a cyclophosphamide derivative) and steroids in addition to cyclosporin A: 5 of these died of infections and 1 also had a lymphoma. 11 patients received additional steroids: 1 of these died from septicaemia and lymphoma. Nephrotoxicity can be avoided by perioperative hydration and forced diuresis. Cyclosporin A is effective on its own and is a very potent immunosuppressive drug. Additional immunosuppressive agents may lead to severe complications.


Timing of cyclosporin-A therapy for abrogation of HVG and GVH responses in rats. Treatment with cyclosporin A was most effective in abrogating popliteal-lymph-node enlargement induced by host-versus-graft and graft-versus-host reactivity in rats when started before injection of donor-strain lymphocytes. Popliteal lymph-node enlargement was never completely abolished, and splenic lymphocytes from recipients treated with cyclosporin A showed no significant reduction in their response to donor-strain lymphocytes in mixed lymphocyte cultures, suggesting that clonal deletion had not taken place. Mixed lymphocyte cultures also indicated that cyclosporin treatment had not reduced the antigenicity of recipient lymphocytes towards donor strain.


Non-surgical treatment of biliary obstruction. Bileduct catheterisation percutaneously through the liver can be used in patients with obstructive jaundice as an adjunct or as an alternative to surgery. Preoperative drainage allows adequate treatment of severe cholangitis and reduces jaundice. Palliative drainage, whether internal or external, can be used instead of surgery. Drainage through the liver succeeded in 40 of 41 patients. Two complications followed the procedure and were treated conservatively. Bile drainage was established through an endoprosthesis into the duodenum in 7 patients and externally through a catheter in the remaining 33. The technique is described, and its use in patients with suppurative cholangitis and benign and malignant biliary strictures is illustrated.


Retrograde cerebral embolism.

Retrograde cerebral embolism. Cerebral embolism may complicate lesions of the subclavian arteries or aortic isthmus distal to the origins of the vertebral or carotid arteries. This may be due to retrograde propagation of occlusive thrombus. In other cases the vessels are patent but there are periods of reverse blood flow, creating a potential for reflux of embolic material to the ostia of the neck vessels.


An erythrocyte membrane-protein anomaly in march haemoglobinuria. In three patients with march haemoglobinuria a well-defined protein abnormality has been revealed by high-resolution sodium-dodecyl-sulphate/polyacrylamide-gel electrophoresis.


Factor VIII-related antigen (VIII R:AG) in haemophilic patients and in carriers. All of fourteen patients with severe classic haemophilia and twelve of fifteen obligate carriers had a pre-peak ("rocket") above the sample well when factor VIII-related antigen was examined by crossed immunoelectrophoresis. The pre-peak may be found in other conditions--including von Willebrand's disease--but if these conditions can be excluded, the pre-peak may be helpful in identification of carriers or haemophilia.


Do human B and null lymphocytes form a single immunoglobulin-bearing population? Human "null" lymphocytes have been recognised because they lack T-lymphocyte membrane markers and the surface membrane immunoglobulin (smIg), detectable by direct immunofluorescence, which conventionally characterises B lymphocytes. But almost all null lymphocytes express smIg determinants which are demonstrable by the direct antiglobulin and mixed antiglobulin rosette tests. It is argued that the definition of a B lymphocyte should be extended to include the null cells which react in these tests. If the heterogeneity of the B-lymphocyte population is more widely recognized, the precise nature of human natural and antibody-dependent killer cells may become clearer.


Effects of burglary and vandalism on the health of old people. The stress of burglary and vandalism can precipitate a major health crisis in old age that demands urgent admission to hospital. Despite reassurance and appropriate treatment, many patients never regain enough confidence to return home.


Reduction in sudden deaths by a multifactorial intervention programme after acute myocardial infarction. 375 consecutive patients below 65 years who had an acute myocardial infarction (AMI) took part in a randomised rehabilitation and secondary prevention trial (part of a W.H.O.-coordinated project) designed to study the effects of a multifactorial intervention programme on morbidity, mortality, return to work, &c. After three years' follow-up the cumulative coronary mortality was significantly smaller in the intervention group than in the controls (18.6% versus 29.4%, p = 0.02). This difference was mainly due to a reduction of sudden deaths in the intervention group (5.8% versus 14.4%, p less than 0.01). The reduction was greatest during the first six months after AMI. 18.1% in the intervention group and 11.2% in the controls (p less than 0.10) presented with non-fatal reinfarctions. The number of patients with new Q-QS findings at the end of the three years was, however, almost the same in both groups. The results suggest that organised aftercare during the first six months after AMI with special emphasis on optimum medical control and health education contributes significantly to a reduction in the number of sudden deaths.


Comparison of accuracy of 99mTc-pyridoxylidene glutamate scanning with oral cholecystography and ultrasonography in diagnosis of acute cholecystitis. A prospective study of 116 patients admitted as emergencies with a clinical diagnosis of acute cholecystitis or biliary colic has shown that the best investigation for confirming a diagnosis of acute cholecystitis is 99mTc-pyridoxylidene glutamate (PG) scanning. Its sensitivity is 99% and its specificity 86%, whereas those of oral cholecystography are 75% and 82%, respectively, and those of ultrasonography are 54% and 62%, respectively. However, estimation of plasma liver enzymes is essential to exclude acute hepatitis before proceeding to early cholecystectomy.


IgA immune complexes in Henoch-Schönlein purpura. Raised levels of IgA immune complexes were found in 13 of 18 children with Henoch-Schönlein purpura irrespective of whether they developed nephritis or not. In contrast, only those children who developed nephritis had raised levels of IgG immune complexes as well. In 2 children with nephritis, two discrete peaks of IgA complexes were found, corresponding to 4 x 10(5) to 8 x 10(5) and 2.5 x 10(6) to 4 x 10(6) daltons. IgG complexes were found only in the larger-molecular-weight peak. Such differences in immune complexes may underlie the different manifestations of this disease.


Association between pre-transplant natural kill and graft-versus-host disease after stem-cell transplantation. Natural killer activity against herpes simplex virus type 1 infected fibroblasts NK(HSV-1) was studied prospectively in patients undergoing allogenic bone-marrow or fetal-tissue stem-cell transplantation. Thirteen patients showed evidence of engraftment and survived long enough to develop graft-versus-host disease (GvHD). Of this group, all of the seven having normal NK(HSV-1) activity before transplantation acquired GvHD and the six having low NK(HSV-1) had no evidence of GvHD. These results were independent of mode of preparation of patients for transplantation, source of stem cells used, or cytomegalovirus infections, and they suggest that this assay reflects a host-determined function capable of stimulating GvHD.


Abnormal intestinal permeability to sugars in villous atrophy. Intestinal permeability to a monosaccharide and a disaccharide was compared by simultaneous measurement of the urinary excretion of L-rhamnose and lactulose after oral ingestion of an hypertonic solution containing both sugars. Urine samples were analysed for sugar content by quantitative thin-layer or paper chromatography. Results in thirteen patients with untreated villous atrophy were compared with those in twelve healthy volunteers. Urinary L-rhamnose excretion was significantly decreased (-40%, p less than 0.02) in patients with villous atrophy, whereas lactulose excretion was paradoxically and significantly increased (+345%, p less than 0.01). The median value of the lactulose/L-rhamnose urinary excretion ratio was sevenfold higher in the patients with villous atrophy; there was no overlap of values for patients and volunteers (p less than 0.01). It is postulated that reduced L-rhamnose urinary excretion in untreated villous atrophy is due to a decreased absorptive area in the small bowel, whereas increased lactulose excretion indicates leakiness of the abnormal mucosa to larger polar molecules.


Hepatic angiosarcoma associated with androgenic-anabolic steroids. A retrospective epidemiological study of deaths from hepatic angiosarcoma (HAS) in the U.S. showed that during 1964--74 there were 168 such cases, of which 37 (22%) were associated with previously known causes (vinyl chloride, 'Thorotrast', and inorganic arsenic) and 4 (3.1%) of the remaining 131 cases with the use of androgenic-anabolic steroids. It is suggested that the long-term use of androgenic-anabolic steroids is the fourth cause of HAS, the majority of cases still being of unknown aetiology. Moreover, the presented cases serve as a link in a spectrum of hepatic disorders recently recognised to be caused by environmental agents such as vinyl chloride, arsenic, and thorotrast, and by contraceptive and anabolic steroids. Similar precursor stages, usually not recognised by clinical laboratory tests and consisting of areas of hyperplasia of hepatocytes and sinusoidal cells and sinusoidal dilatation, lead potentially to hepatic adenoma, carcinoma, peliosis, and angiosarcoma.


Fetal iodine deficiency and motor performance during childhood. Motor performance of children born to mothers living in an iodine-deficient region was assessed. The mothers were participants in a controlled trial of intramuscular iodised oil in the prevention of endemic cretinism carried out in the Western Highlands of Papua New Guinea. Mothers received either iodised oil or placebo saline. Children born to mothers given iodine were significantly faster and more accurate in tests of manual function than children from control mothers. The findings indicate that iodine deficiency may lead to a spectrum of subclinical deficits which place the children at a developmental disadvantage.


Chloroquine-resistant Plasmodium falciparum from East Africa: cultivation and drug sensitivity of the Tanzanian I/CDC strain from an American tourist. A strain of Plasmodium falciparum, designated Tanzanian I/CDC, from an American tourist returning from Tanzania, was isolated in vitro and in the Aotus monkey. Clinically, the infection showed a late recrudescent pattern of chloroquine resistance. In 2 inoculated Aotus monkeys, the infection recrudesced after a dose of chloroquine (40 mg/kg) curative for sensitive P. falciparum strains in the Aotus monkey. In 4 ,dditional monkeys two primary infections and one of the recrudescent parasitaemias were cured with a 100 mg/kg dose of chloroquine; the second recrudescent parasitaemia was cured with an additional 40 mg/kg dose of chloroquine. The 48 h in-vitro chloroquine-sensitivity test demonstrated that the Tanzanian I/CDC strain had a pattern of chloroquine resistance similar to a reference resistant strain, the Vietnam-Oak Knoll (FVO). These studies reinforce reports which suggest that chloroquine-resistant malaria is being transmitted in East Africa.


Site of Epstein-Barr virus replication in the oropharynx. Cells and cell-free material in saliva, parotid secretions, and throat washings from patients with acute infectious mononucleosis and patients undergoing tonsillectomy were assayed for the presence of infectious Epstein-Barr (EBV) virus. The agent was invariably present in cell-free form in saliva; neither infectious virus nor viral antigens were found in the cells. Tonsillar lymphocytes from eight patients were also free of EBV. In 10 of 40 patients virus was recovered in secretions from parotid-gland orifices or ducts. These findings suggest that the salivary glands are the site of EBV production in the oropharynx.


Prevention of effects of alcohol intoxication by naloxone. In a double-blind, cross-over study of twenty male volunteers intravenous injection of 0.4 mg naloxone prevented the impairment of psychomotor performance induced by low levels of blood alcohol. The possibility that alcohol produces intoxication by stimulating the release of endogenous opioid peptides should be investigated.