Vibriolytic IgG immunocyte response of mice after primary and secondary immunization with cholera somatic antigens. Antibody plaque-forming cells (FC) to the somatic antigens of Vibrio cholerae were enumerated in the spleen of mice after primary and secondary immunization with a heat-killed vaccine prepared from the vibrios. Immunocytes releasing both high efficiency IgM and low efficiency IgG antibody were readily detected using a direct and facilitated plaque procedure in agar gel. Whereas the peak numbers of IgM-PFC after primary immunization occurred on days 12 to 14, the peak IgG-PFC response developed somewhat later (16-18 days). After a second injection of vaccine larger numbers of both IgM- and IgG-PFE appeared in the mouse spleens, with peak responses for both occurring between days 5 and 8. The largest number of IgG-PFC developed in spleens of mice given a second injection of vaccine 6-8 weeks after primary immunization. The dose of killed vibrios used for priming markedly affected both the magnitude and the class of antibody-forming cells appearing during the secondary response; 1--10 mug vaccine was more effective than higher or lower doses for priming the mice to a heightened secondary response. Furthermore, the antigenic specificity of both the IgM- and IgG-PFC appearing after secondary immunization was directly related to the strain of cholera bacilli used for priming. When mice were immunized with the Ogawa strains of cholera most of the secondary PFC after booster immunization with the serologically distinct Inaba strain was directed towards the common antigen shared by both strains and not to the type specific antigen of the Inaba vibrios. The specificity of the anti-vibrio PFC during both the primary and secondary responses was readily demonstrable by inhibition experiments using sonicated or soluble cholera antigens. Prior incubation of these antigens with test spleen cells in the agar gel effictively inhibited development of the vibriolytic plaques, regardless of antibody class. Similar antigen extracts from toher bacteria had no effect. The immunoglobulin nature of the plaques was also demonstrable by inhibition with low dilutions of rabbit anti-mouse globulin serum incorporated into the agar plates prior to testing; both IgM and IgG plaues were inhibited.


Studies on the control of antibody synthesis. VIII. Selection for high affinity antibody production in the secondary response. Priming with haptenic determinants can augment the antibody response to a new haptenic determinant presented on a different carrier to which the original haptens are also coupled. This augmentation may be accompanied by a slight increase in the affinity of the antibody formed. However, only if the test hapten is present on the original priming antigen is there a selection for high affinity antibody-forming cells and a marked increase in the affinity of the antihapten antibody produced upon boosting is seen. The data are consistent with the assumption that antibody affinity is primarily controlled by the selection of B lymphocytes by antigen, while additional factors may be involved in controlling the magnitude of the response.


Evidence for a membrane carrier molecule common to embryonal and tumour-specific antigenic determinants expressed by a mouse transplantable tumour. Rabbits were primed with membrane antigens solubilized from BALB/c embryo cells. After boosting with membrane antigens solubilized from a syngeneic transplantable adenocarcinoma, they developed a 'secondary' response against tumour-specific antigenic determinants. The antibodies against these determinants neither reacted with nor were absorbed by the antigens prepared from embryonal cells. However, the antigen displaying the tumour-specific determinants was bound by a reversed immunoadsorbent of insoluble anti-embryo antibodies. Indirect immunofluorescence experiments performed on adenocarcinoma cells in culture showed that, under conditions where redistribution of cell membrane components was induced, the anti-embryo antiserum aggregated the tumour-specific determinants. The purification of embryo and tumour-specific antigens achieved by affinity chromatography on insoluble antibody columns yielded three polypeptides of molecular weight close to 25,000, 20,000, and 10,000 Daltons respectively. It is suggested that the antigenic determinants responsible for tumour and embryo specificities in adenocarcinoma were located on the same molecule, or, more likely, on molecules which are closely associated in the plasma membrane and that do not dissociated in bile salts.


Inhibition of IgE and compound 48/80-induced histamine release by lectins. Lectins from Ricinus communis and Glycine max, as well as wheat germ agglutinin and concanavalin A, caused a dose-dependent release of histamine from mast cells present in the mixed peritoneal cells from the rat. In addition, histamine release in an IgE-mediated and a compound 48/80-mediated reaction was inhibited in cells which had been pretreated with these lectins. With concanavalin A and the R. communis lectin both effect were prevented by the addition of the appropriate monosaccharides to the incubations. However, the lectin-induced histamine release and the lectin-induced inhibition of subsequent IgE-mediated histamine release could be dissociated: thus L-rhamnose, a hexose not ordinarily found on mammalian cell membranes, a specifically inhibited histamine release which was caused by the lectin from R. communis without affecting the inhibition of IgE-mediated histamine release. Conversely, D-fucose, which also is not a constituent of cell membrane glycolipids or glycoproteins prevented the inhibition of IgE-mediated histamine release by this lectin without affecting the lectin-induced histamine release. Furthermore, the nominally galactose-specific lectins from Sophora japonica and Ulex europeus inhibited IgE-mediated histamine release while causing little if any histamine release themselves. High concentrations of the lectin from Lotus tetragonolobus failed to cause histamine release or to affect the IgE-mediated histamine release reaction. Based on the known structural specificity of these lectins and the amounts of the lectins which were required to demonstrate an effect, it was concluded that D-galactose, alpha-linked, intrachain D-glucose (or mannose), and N-acetylglucosamine residues but probably not N-acetyl-galactosamine or L-fucose residues in the glycolipids or glycoproteins of the mast cell membrane can play a role in the initiation of histamine release and in the desensitization of the cells to subsequent histamine release-inducing stimuli.


Trichinella infestation in mice genetically selected for high and low antibody production. Mice genetically selected according to their capacity to produce antibodies (high and low responder lines) were infested with Trichinella spiralis. High responder mice produced much higher levels of IgE and gamma1 serum antibodies than low responder mice. IgG and IgM antibodies were detected by immunodiffusion analysis or passive haemagglutination only in animals of the high responder line and in very low concentrations. The number of living larvae present in the muscles was very similar in the animals of both lines 30, 60 or 75 days after infestation.


Antigenic heterogeneity among pyocins of Pseudomonas aeruginosa. We investigated the effect of FeSO4 on phagocytosis-associated, increased oxidative metabolism via the hexose monophosphate shunt, with special attention to its effect on H2O2 levels. The availability of glutathione peroxidase and glutathione reductase for H2O2 disposal and hexose monophosphate shunt stimulation also are evaluated. The results show an impairment of phagocytosis-associated hexose monophosphate shunt activity together with an increase both of resting and phagocytosing formate oxidation. These apparently paradoxical findings are resolved by demonstrating a direct enhancement of formate oxidation by FeSO4 in a cell-free system. In addition, measurement of H2O2 concentrations via scopoletin fluorescence shows reduction of H2O2 by FeSO4. There is no effect on either glutathione peroxidase or glutathione reductase activities. These data suggest that one mechanism of FeSO4 impairment of microbicidal activity is by its removal of H2O2.


Evidence for O antigens as the antigenic determinants in "ribosomal" vaccines prepared from Salmonella. The nature of the protective substance in ribosomal ribonucleic acid and protein extracts of Salmonella has been investigated. The results of experiments in which vaccines were prepared from isogenic strains and strains with defects in lipopolysaccharide synthesis show that O antigens contaminate both ribonucleic acid and protein ribosomal extracts, and are responsible for at least part of their strain-specific protective activity. In addition, it was observed that a ribosomal ribonucleic acid preparation from S. adelaide contains a heat-stable immunogen which is not an O antigen or that gives cross-protection across species lines. The contribution of ribosomes to the immunity induced by "ribosomal vaccines" is discussed.


Inhibition of immediate hypersensitivity reactions in the rat by ICI 74,917 and disodium cromoglycate. ICI 74,917, a potent inhibitor of IgE-mediated passive cutaneous anaphylaxis (PCA) in the rat, exhibited tachyphylaxis in that pre-dosing sensitised rats with a high dose of compound reduced the inhibitory effect on rat PCA of a second dose given at challenge. This phenomenon was most apparent when the pre-dose-challenge dose interval was 15--60 min. Similar findings were obtained using antigen-induced histamine release in vitro from rat peritoneal cells. In these respects, ICI 74,917 was similar to disodium cromoglycate (DSCG) although DSCG appeared less effective in inducing tachyphylaxis than ICI 74,917. There was no evidence in vivo or in vitro that a high dose of either DSCG or ICI 74,917 enhanced the activity of a second low dose of either drug given at challenge.


Penicillin hypersensitivity in systemic lupus erythematosus. The prevalence of positive immediate-type skin tests to penicillin antigens among patients with systemic lupus erythematosus (SLE) was compared with hospitalized controls without SLE and found not to differ significantly. On the other hand, we confirmed previous reports that the prevalence of histories of prior penicillin reaction was significantly higher in SLE patients than among controls. This is felt to be related to the primary disease process, and we conclude that patients with SLE do not have an increased risk of IgE-mediated allergic reactions to penicillin.


Binding of human IgE immunoglobulin to rat mast cells. A study by means of three independent techniques. The binding of human myeloma IgE immunoglobulin on rat mast cells was studied by three independent techniques. A mixed agglutination reaction with anti-IgE-coated Sephadex granules demonstrated that only human IgE-coated rat mast cells were clearly agglutinated. This binding is strong (50% agglutination) in 3 min and progresses for 30 min (95% agglutination). Autoradiographic studies with 125I-labelled human serum proteins demonstrated the selective formation of grains on mast cells incubated with labelled IgE. Upon action of anti-IgE antiserum on IgE-coated rat mast cells, the mast cells released up to 47.5% of their total histamine content in a fluorometric histamine assay. A relationship was established between sensitizing doses of human IgE and histamine release. These results bring evidence for a binding of human IgE on rat mast cells and imply the existence of receptors for this immunoglobulin on mast cell membrane.


Specificity of antigens in aqueous phenol extracts of skin examined by means of guinea pig and rabbit immune sera. Human, guinea pig and rabbit skin homogenates were digested with trypsin and extracted with phenol water. Antisera were raised in guinea pigs and rabbits by immunization with extract recovered from the water phase (TPW extract). All sera showed increased titres in indirect haemagglutination tests. The results of absorption and inhibition experiments indicated antibodies against a common antigenic determinant. These antibodies also agglutinated erythrocytes sensitized with autologous antigen. In addition, serum from rabbits immunized with human or guinea pig skin extract contained antibodies against species-specific determinants. Rabbit antiserum precipitated guinea pig skin extract. The antigen involved had specificity identical with that of an antigen in the human, but not in the rabbit skin, extract. Oxidation of the human TPW extract with periodate destroyed the precipitinogen and the species-specific haemagglutinogen while the common determinant was not affected.


Restoration of specific immunity against SV40 tumor-specific transplantation antigen to lymphoid cells from tumor-bearing mice. Specific cell-mediated immunity to SV40 tumor-specific transplantation antigen (TSTA) in BALB/c mice undergoing progressive tumorigenesis by syngeneic SV40-transformed cells (VLM) was investigated in vivo using a tumor-cell neutralization test. Specific cellular reactivity to SV40 TSTA was not detected in BALB/c mice bearing large tumors (10-15 mm mean diameter) but was demonstrable after tumor excision. Specific cytotoxic reactivity against syngeneic SV40-transformed cells in vivo could be restored to lymphoid cells from VLM tumor-bearing mice either by culturing the lymphoid cells in vitro or by treating them with papain or trypsin. Enzyme-treated lymphoid cells from MCA tumor-bearing BALB/c mice had no cytotoxic reactivity against VLM cells. These studies suggest that tumor-bearing hosts possess lymphocytes which are sensitized to the TSTA of the tumor but that the reactivity of these lymphocytes is blocked.


Innervation of muscle in leprosy with special reference to the muscle spindle. The pattern of extrafusal and intrafusal innervation was studied in muscle biopsies from the flexor carpi ulnaris and biceps brachii muscles, which were clinically unimpaired. Smudginess and enlargement of the motor end plates were the most definitely abnormal feature of the extrafusal innervation pattern; the intrafusal fibers, on the other hand, were either unremarkable or showed increased tortuosity, beading and in more extreme cases of grossly complicated intertwining pattern. Bacilli were frequently found in the spindles, no component being spared. The study emphasizes the significant involvement of striated muscle in leprosy, a fact not revealed by clinical examination alone.


An objective measurement of nonspeaking children's performance with a computer-controlled program for the stimulation of language behavior. A method is described for recording a child's performance with a computer-controlled audio-visual device designed to stimulate language behavior in nonspeaking children. From the recorded data a child's position can be estimated on a curve derived from normal children. This position could be of value in planning treatment for a nonspeaking child and assessing his progress.


Intercellular matrix in colonies of Candida. The histochemistry and fine structure of typical colonies of six species of Candida were studied, using a total of 31 clinical isolates. The colonies consisted of viable and degenerate cells which lay in an intercellular matrix. This matrix was made up of amorphous, granular, and fibrillar components, the relative proportions and total amount of which varied from species to species. The cells of all species were surrounded by a zone of homogeneous amorphus material, which may be a highly cross-linked carbohydrate. This separated intact cells from irregularly distributed granular debris derived from the cytoplasm of degenerate cells. Focal cellular degeneration and associated granular debris were present within the colonies of all species and were most common in the surface layers of cells of colonies of C. albicans and C. tropicalis. The large amounts of intercellular matrix in this region formed a surface coat on colonies of these two species. Intercellular strands of cell wall material, and to a lesser extent other membranous elements from degenerate cells, formed a prominent fibrillar meshwork in the colonies of C. albicans and C. tropicalis, but were less common in those of C. pseudotropicalis and C. guilliermondii and seldom seen in those of C. parapsilosis and C. krusei.


Proteinase C (carboxypeptidase Y) mutant of yeast. A mutant of yeast lacking proteinase C (carboxypeptidase Y) activity has been found by using a histochemical stain to screen mutagenized colonies. This defect segregates 2:2 in meiotic tetrads. Cell extracts lacked the esterolytic, amidase, and proteolytic activities associated with proteinase C. The absence of proteinase C does not affect mitotic growth and has no obvious effect on the formation of viable ascospores or meiotic segregation. The mutant grows on peptides known to be cleaved by proteinase C in vitro. This finding is consistent with the idea that other enzymes exist in vivo with overlapping substrate specificities.


Experimental exophthalmos. Binding of thyrotropin and an exophthalmogenic factor derived from thyrotropin to retro-orbital tissue plasma membranes. Biologically active preparations of 125I-thyrotropin, [3H]thyrotropin, and the [3H]exophthalmogenic factor derived from thyrotropin by partial pepsin digestion have been used to study the binding properties of the thyrotropin receptor on guinea pig retro-orbital tissue plasma membranes. In regard to the optimal conditions of binding, pH, buffer, salt concentrations, and temperature, these properties are the same as those described in any accompanying report concerning thyrotropin binding to bovine thyroid plasma membranes (Tate, R.L., Schwartz, H.I., Holmes, J.M., Kohn, L.D., and Winand, R.J. (1975) J. Biol. Chem. 250, 6509-6515). In addition, thyrotropin receptors on the retro-orbital tissue plasma membranes are similar to thyrotropin receptors on bovine thyroid plasma membranes in their apparent negative cooperativity and in their relative affinities for luteinizing hormone, the beta subunit of thyrotropin, and the alpha subunit of thyrotropin. In contrast, gamma-globulin from patients with malignant exophthalmos enhances binding when added to incubation mixtures containing the retro-orbital tissue plasma membranes but not when added to those containing thyroid plasma membranes. Normal gamma-globulin and gamma-globulin from Graves' disease patients without exophthalmos do not have this property. The gamma-globulin itself does not bind to the membrane except in the presence of thyrotropin or its exophthalmogenic factor derivative. Tryptic digestion of the retro-orbital tissue membranes releases specific thyrotropin and exophthalmogenic factor binding activity into the supernatant phase. Chromatography on Sephadex G-100 indicates that this trypsin-released receptor activity has a molecular weight of 75,000 or greater, rather than 15,000 to 30,000 for the trypsin-released receptor activity from bovine thyroid membranes (Tate, R.L., Schwartz, H.I., Holmes, J.M., Kohn, L.D., and Winand, R.J. (1975) J. Biol. Chem. 250, 6509-6515).


Stimulation of adenylate cyclase activity in retro-orbital tissue membranes by thyrotropin and an exophthalmogenic factor derived from thyrotropin. Retro-orbital tissue membranes have been shown to have adenylate cyclase activity which can be stimulated by thyrotropin and by an exophthalmogenic factor derived from the thyrotropin molecule by partial pepsin digestion. This stimulable activity is maximal after 15 min and is optimal in the presence of 3 mM magnesium and 1.5 mM ATP. Calcium salts are exquisitely inhibitory to the hormonal stimulation; sodium, lithium, and ammonium salts are significantly less inhibitory. Thyrotropin and the exophthalmogenic factor induce similar maximal levels of stimulation but a 4- to 5-fold higher concentration of exophthalmogenic factor is required to achieve this level. Fluoride stimulates adenylate cyclase activity 2- to 3-fold higher than either thyrotropin or the exophthalmogenic factor; thyrotropin, luteinizing hormone, the beta subunit of thyrotropin, and the alpha subunit of thyrotropin have relative activities for stimulation of cyclase activity of 100:2:2 less than 0.5. Several other polypeptide and glycoprotein hormones have no effect. The gamma-globulin from patients with malignant exophthalmos has no significant effect on cyclase activity either alone or in the presence of maximal levels of thyrotropin or the exophthalmogenic factor; this gamma-globulin does, however, stimulate cyclase activity at submaximal hormone levels. Trypsin not only destroys the hormone-stimulable adenylate cyclase activity on retro-orbital tissue plasma membranes, but also destroys it on the 15,000 to 30,000 molecular weight receptor fragment released from the membranes by the tryptic action.


Osteophytes and the osteoarthritic femoral head. The study describes the topography, morphology and growth of osteophytes in forty femoral heads removed from patients presenting with advanced osteoarthritis of the hip. In addition to standard histological techniques, radiography of serial bone slices in vivo bone labelling with tetracycline and 32P were used. The pattern of major osteophyte formation appeared to be influenced by the direction, degree and rate of displacement of the femoral head in relation to the acetabulum; four principal patterns of growth were noted. Osteophytes form part of extensive osteogenic processes that involve bone structure in the osteoarthritic joint.


Mouse-human heterokaryon analysis with a 33258 Hoechst-Giemsa technique. The bibenzimidazol derivative 33258 Hoechst can be used to distinguish microfluorometrically between mouse and human nuclei in heterokaryons.  This affords a quick and accurate alternative to autoradiography in the analysis of such heterokaryons.  The 33258 Hoechst fluorescence patterns can be converted after irradiation to a Giemsa rendition of the differential staining.


Gas-liquid chromatographic studies of reactions and structural relationships of steroids. Part III. 11alpha-hydroxysteroids of the androstane and pregnane series. Qualitative and quantitative effects of classical reactions on steroids observed by gas-liquid chromatography (GLC) under standardized conditions, including the double internal-standard technique, are reported. Simple procedures applicable to nanogram amounts of reactants which afford excellent yields of the major products are described. Reactions studied include the Wolff-Kishner removal of keto groups, their conversion into hydroxyl groups with sodium-ethanol or sodium borohydride and into dioxolone derivatives with ethylene glycol; the conversion of hydroxyl into keto groups with chromium trioxide and to trimethylsilyl (TMS) ethers by hexamethyldisilazane; the hydrolysis of dioxolone and TMS derivatives by H+. Gas-liquid chromatograms of reaction mixtures of single- and multistep reactions readily provide information on the effects on the 11alpha-hydroxy and other functional groups at positions 3 and 17 (androstane series) and positions 3 and 20 (pregnane series), and the retention times of many steroids unavailable from commercial or other sources. GLC data analysis provides relationships between steroid structure and retention time from which methods for the computation of retention times and for steroid identification are designed. The accuracy of the calculation methods is demonstrated.


Detection of antithyroid residues in meat and some organs of slaughtered animals. A simple method is described for the routine detection of antithyroid residues in thyroid, liver, kidney and meat contaminated at levels as low as 10 ppb (10 parts per 10(9)). Tissue samples (2 g) are homogenized in methanol, contaminating lipids and amino acids are removed and the antithyroid residues are subjected to reaction with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) in buffer. The NBD derivatives are extracted with diethyl ether and separated by thin-layer chromatography. After spraying with cysteine or mercaptoethylamine, the antithyroid residues appear as fluorescent spots. The detection limit of these compounds is of the order of 200 pg.


Hyperthyroidism in Tasmania following iodide supplementation: measurements of thyroid-stimulating autoantibodies and thyrotropin. Serum thyroid-stimulating autoantibodies (LATS and LATS protector) and thyrotropin (TSH) concentrations were measured in the serum of 30 patients with hyperthyroidism living in Tasmania who developed their disease following correction of iodine deficiency by addition of iodate to the bread. Patients were grouped according to thyroid scan results. None of 8 patients with autonomous thyroid nodules had thyroid-stimulating autoantibodies. These were present in both of the patients with uniform thyroid scans and 14 of 20 patients (70%) with irregular scans without demonstrated localized autonomy. Serum TSH, measured by immunoassay of concentrated serum extracts, was 0.15 muU/ml or less in all patients, below the range of 0.35 to 2.60 muU/ml found in normal subjects. Only  6 (20%) of the 30 patients failed to show either localized autonomy or thyroid-stimulating autoantibodies. In most regards these patients resembled those with antonomous nodules. The findings support the conclusion that the increased incidence of phyerthyroidism in Tasmania was due to an increased supply of iodine to patients with latent hyperthyroidism whose thyroid glands, due to the presence of toxid nodule(s) or thyroid-stimulating autoantibodies, were unresponsive to control by TSH deprivation. There was no evidence for additional pathogenic mechanisms


Daytime titers of testosterone, LH, estrone, estradiol, and testosterone-binding protein: acute effects of LH and LH-releasing hormone in men. Four normal 18-20 yr-old men were studied on 3 occasions, from 0830 h to 1500 h. The baseline for each study consisted of 3 or 4 measurements of the respective hormone obtained between 0830 and 0900 h. In the control studies mean testosterone (T) fell by 43% (P less than 0.01) during the final 30 min. The fall was gradual throughout the day and was significant by 1100 h  (P less than 0.05). Administration of LH and LH-releasing hormone (LHRH) at 0900 h resulted in 9-fold (5 min) and 3-fold (30 min) higher concentrations of LH respectively. LH declined more slowly after LHRH. Titers of T rose to the 0830-0900 h mean 130 min after LH but were never significantly elevated; the occurrence of a significant drop in mean T was delayed for 70 min. After  LHRH there was a nonsignificant 24% increase of the mean T followed by a slow decline; however, T did not fall significantly below the mean baseline level. In contrast, in 2 of the 4 subjects LHRH resulted in rises in T levels (P less than 0.05) above the basal titers. Testosterone-binding globulin (TeBG) mean titers showed no diurnal rhythm in the control studies. There were statistically significant elevations of mean TeBG 150 min after LH and 340 to 370 min after LHRH, as well as sustained increases during the final 30 to 210 min of 1 or 2 individuals in each group. The reason for these increases in TeBG is not presently known. Estrogen analyses performed in all studies on 2 of the subjects revealed: 1) afternoon titers of estrone were lower than baseline in all 6 studies, 2) there was no diurnal rhythm for estradiol in control studies, and 3) estradiol increased during the final 2.5 to 3 h after LHRH (P less than 0.01), but after LH it was not altered in 1 man and was lower in the other.


Thyroidal accumulation of 131I during adrenal gland scintigraphy with 131I-19-iodocholesterol: effects of thyroid blocking agents. 131I-19-iodocholesterol has been used by Beierwaltes et al., to visualize human adrenal glands. The tissue distribution of 131I-19-iodocholesterol was determined by Morita et al., both in dogs and men. No data have been reported on thyroidal 131I accumulation after administration of the radiopharmaceutical. We calculated the percentage uptake of radioactivity in the human thyroid gland inhibited with various therapeutic agents (triiodothyronine (T3), Lugol's solution, potassium perchlorate (KC1O4).


Stimulation of human neutrophils by soluble and insoluble immunoglobulin aggregates. Secretion of granule constituents and increased oxidation of glucose. Reaction of human neutrophils with aggregated immunoglobulin on nonphagocytosable surfaces results in secretion of granule enzymes (exocytosis of granules) and stimulation of glucose oxidation by the nexose monophosphate pathway (HMP). The role of HMP stimulation in the enzyme secretion and some requirements for the two neutrophil activities have been examined. It was found that (a) HMP stimulation could be selectively inhibited under conditions  where release of granule enzymes remained unchanged or was enhanced, for example, by reduced glucose concentration or by 2-deoxyglucose. (b) Removal of Ca++ and addition of agents which increased the intracellular levels of cyclic AMP (cAMP), however, prevented both activities, while colchicine had greater inhibitory activity on HMP stimulation than upon secretion. (c) Neutrophils incubated in suspension with particulate aggregated gamma-globulin phagocytosed the particles and exhibited a stimulated HMP and released granule enzymes. In contrast, incubation in suspension with soluble aggregated gamma-globulin resulted in the stimulated HMP only. Granule evzymes were not liberated. 300-fold less soluble aggregates bound to a surface, however readily induced exocytosis of granules from adherent neutrophils. This demonstrates the importance of surface effects in the induction of secretion from neutrophils. Aggregated immunoglobulin reacting with neutrophil Fc receptors thus induces both degranulation (exocytosis) and increased HMP activity. The pathways leading to these events are separable although apparently sharing some common steps, including the initiating events.


Selective subcellular localization of cations with variants of the potassium (pyro)antimonate technique. Fixation of rat parotid with an unbuffered osmium tetroxide solution containing nearly saturated potassium (pyro)antimonate resulted in abundant deposition of cation-antimonate precipatates in acinar cells. Altering the antimonate concentration, including buffers or chelators in the solution or changing the primary fixative resulted in an altered intensity and distribution of the precipitates formed in the tissue, apparently reflecting a degree of selectivity in ion localization. Decreasing the concentration of pyroantimonate to about half-saturation preserved predominantly the less soluble antimonate salts (e.g., Na+, Ca++) and resulted in preferential retention of deposits along the plasmalemma and in mitochondrial "dense bodies," with loss of most cytoplasmic and nuclear precipitates. A similar pattern was seen if fixation with the high concentration antimonate-osmium procedure was followed by a prolonged rinse. Adding phosphate or collidine buffers markedly decreased precipitates in the nuclei and on granular reticulum as well. Phosphate buffer or ehtyleneglycoltetraacetate inhibited in vitro precipitation of calcium and sodium and decreased or abolished plasmalemmal deposits. Glutaraldehyde fixation, either in the presence of antimonate or prior to antimonate-containing osmium tetroxide, abolished heterochromatin deposits. Mitochondrial dense bodies were of two types, one containing precipitate and the other inherently osmiophilic. The latter were also observed in pyrophosphate-osmium controls. Results from in vitro titrations of cations with the various antimonate methods and from neutron activation analyses of fixed tissues supported conclusions drawn from fine structural distribution patterns and were interpreted as follows. In rat parotid acinar cells, deposits in heterochromatin and on granular reticulum probably arose from precipitation in sites of high K+ and H+ as well as--NH3+-rich histones. Plasmalemmal antimonate deposits demonstrated sites of sodium and/or calcium accumulation. Some mitochondrial dense bodies contained Ca++ whereas others were inherently osmiophilic. Large, extracellular deposits were probably predominantly sodium precipitates.


Cytochemical localization of peroxisomes in Tetrahymena pyriformis. Pronounced staining of Tetrahymena pyriformis peroxisomes was demonstrated when glutaraldehyde-fixed cells were incubated in an alkaline medium containing 3,3'-diaminobenzidine and hydrogen peroxide. A variable amount of electron-opague deposit was observed when cells were incubated in diaminobenzidine and H2O2 for 1 hr while an intense deposit followed incubation for 4 hr in the same medium. The staining was abolished completely when 3-amino-1,2,4-triazole, KCN or NaN3 was added to the incubation medium. Based on these cytochemical observations and the morphologic identification by size, shape and other ultrastructural details, it is suggested that this study presents evidence for a conclusive morphologic identification of Tetrahymena peroxisomes.


Ultrastructural localization of gamma-chain and immunoglobulin G in human lymphocytes using enzyme-labeled Fab fragment. By the use of rabbit antibodies against the heavy chain of human immunoglobulin G (IgG), the gamma-chain and IgG molecules were successfully localized at the ultrastructural level in human peripheral lymphocytes. The rabbit Fab fragment was coupled to horseradish peroxidase by means of glutaraldehyde and the resulting conjugate could penetrate the intact plasma membrane. Discernible reaction product was observed in cisternae of the nuclear envelope, rough endoplasmic reticulum and Golgi apparatus as well as on the surface of the lymphocytes. In normal human individuals under no specific antigenic stimulation, only a few peripheral lymphocytes showed a rare positive intractoplasmic reaction. Reaction product may represent either the whole IgG molecule, the half molecule consisting of one heavy and one light chain or nascent gamma-chain.


Occurrence of viral hepatitis type B and type non-B in three patients. Acute viral hepatitis type B was observed in three patients convalescing from hepatitis B surface antigen (HBs Ag)-negative viral hepatitis. The two types of viral hepatitis were clearly differentiated by serial HBs Ag determinations with counterimmunoelectrophoresis and radioimmunoassay. These findings provide additional evidence of the lack of cross-immunity between the two types of viral hepatitis and indicate that patients with HBs Ag-positive and HBs Ag-negative acute viral hepatitis should be separated.


Exit block in sustained atrial tachycardia in two children. The electrocardiographic findings in two children with sustained atrial tachycardia and exit block are reported. The exit block was dependent on the atrial pacemaker rate, being present only at rapid rates of discharge. In one case propranolol enhanced the exit block, and on an appropriate dose the heart rate was maintained within a well tolerated range.


Serological relationship between a pathogenic strain of Marek's disease virus, its attenuated derivative and herpes virus of turkeys. Precipitating antigens present in extracts of chick embryo cells infected with the HPRS-16 attenuated strain of Marek's disease virus (att-MDV) were separated by gel filtration on Sephadex G200 and some of their properties determined. The two main antigens detected with convalescent MD serum, referred to as 'B' and 'C' antigens, had mobilities of 0-55 and 0-25 respectively relative to phenol red on electrophoresis in 7-5% acrylamide gel. The B antigen was relatively stable and of low mol. wt. in comparison with the C antigen. B and C antigens were in some instances also detected in culture medium of infected cells, but were distinguishable from the A antigen, a major glycoprotein antigen released into the culture medium of cells infected with HPRS-16. The results of immunodiffusion studies suggest that B antigen is common to MDV and strains of herpes virus of turkeys(HVT) and that at least 2 antigens (including C) are MDV specific. The A antigen was also common to MDV and HVT strains. It was noted however that the capacity of HPRS-16/att to synthesize A antigen was considerably reduced in comparison with HPRS-16 and HVT strains, and in some preparations the A antigen could not be detected. Evidence was also obtained for the presence of HVT-specific antigens associated mainly with the cell fraction.


Cellular immune response to peripheral nerve basic protein in idiopathic facial paralysis (Bell's palsy). Lymphocytes from patients with Bell's palsy were shown to undergo significant stimulation when cultured in vitro in the presence of a purely neuritogenic basic protein (P1L) isolated from human peripheral nerve myelin. No sensitization was observed to other neural antigens, namely, another periperal nerve myelin basic protein (P2) and the central nerve myelin basic encephalitogenic protein (BE). A similar pattern of response was also demonstrated in patients with Guillain-Barré syndrome (GBS). Lymphocytes from patients suffering from other neuropathies or other diseases involving the face showed no response to any of these antigens. The specific in vitro response to P1L protein in Bell's palsy may suggest that an in vivo sensitization of lymphocytes to such self protein occurs in this condition, and that cell-mediated, probably post-infectious, autoimmune mechanisms may be an important factor in the pathogenesis of the paralysis. Thus, Bell's palsy is immunologically similar to GBS, or may even represent a mononeuritic variant of GBS. In view of these findings the administration of steroids to patients with Bell's palsy seems logical on the basis of their immunosuppressive action.


Monocytes and histiocytes in cell cultures of cerebrospinal fluid. Morphology of cultured CSF cells. A method of CSF cell culturing, based on observations of cultured cells isolated from 700 CSF specimens obtained for routine diagnostic procedures by lumbar puncture from patients who had no proven or suspected neoplastic disease, is described which enables the demonstration of proliferating mononuclear elements even when they are present in specimens with low cell count. Spread on surfaces of plastic and glass material, monocytes and histiocytes in CSF cell cultures can appear as polygonal or crescent shaped epitheloid cells, may assume spindle shapes, or transform into multinucleated giant cells. Some cells given rise to clones with different rates of proliferation, up to the formation of a monolayer. After short term culturing the cytochemical characteristics of the cells are comparable to those of the native cells. Phagocytosis in culture is possible. Cells with a high rate of proliferation can be isolated from CSF specimens in subacute non-bacterial inflammatory processes, in chronic meningitis, in the state of repair of bacterial meningitis and subarachnoid hemorrhage, after repeated lumbar punctures and other unspecific irritations such as myelography and pneumencephalography, and in the course of intrathecal cytostatic therapy.


The effect of phenytoin and ethosuximide on primidone metabolism in patients with epilepsy. Little is known about the influence of phenytoin and ethosuximide on primidone. Therefore we studied three groups of patients: 28 receiving primidone alone, 16 on comedication of primidone with phenytoin and 9 on primidone plus ethosuximide. Antiepileptic drug determinations were done with Kupferberg's gas chromatographic method. The results show that the addition of phenytoin--but not ethosuximide--does increase the plasma concentration of phenobarbital derived from primidone but not of primidone itself. The phenobarbital/primidone plasma concentration ratio is with 4.2 +/- 0.7 (+/- S.E.) significantly (P less than 0.001) higher in patients receiving primidone and phenytoin as compared to those on primidone alone (1.6 +/- 0.2) or together with ethosuximide (1.4 +/- 0.7). The effect of phenytoin occurs and persists for several days after the steady state plasma concentration of phenytoin has been reached. This effect is probably not due to induction of enzymes hydroxylating primidone but rather to inhibition of the metabolism and/or excretion of phenobarbital. A case of phenobarbital intoxication due to addition of phenytoin to primidone medication is described in detail.


The accessory deep peroneal nerve.

The accessory deep peroneal nerve. The innervation of the right M. extensor digitorum brevis was investigated in 52 subjects with the aid of stimulation electromyography. In 13 cases (25%) a particular branch of the superficial branch of the peroneal nerve, the accessory deep peroneal nerve, could be identified. In 1 case it alone, and in 12 cases, together with the deep peroneal nerve, innervated the M. ext. dig. brevis. The significance of the awareness of this anomalous innervation is pointed out.


[Peripheral nerve compression by ganglionic cysts (author's transl)]. Extra- and intraneural ganglionic cysts rarely involved peripheral nerves. They are found in the neighbourhood of large joints. Intraneural cysts prefer the deep peroneal nerve and cause intermittent pain and severe nerve damage. The ulnar nerve is affected most often at the wrist. There are different types of distal motor and/or sensory ulnar palsy. Spontaneous recovery may take place and recurrences of intraneural cysts of the peroneal nerve occur after surgery.


[Subacute myelo-optic-neuropathy (S.M.O.N.) following treatment with clioquinol (author's transl)]. 2 patients, who were treated with clioquinol after radical resection of carcinoma of the rectum and colostomy, developed symmetrical sensorimotor polyneuropathy, mild posterior tract ataxia, bilateral pyramidal tract lesions and optic neuropathy, a clinical picture compatible with subacute myelo-optic-neuropathy (S.M.O.N.). One patient had neurological symptoms after having received 750 g of clioquinol, 3 years after treatment started, and impairment of vision was noted after having received 1200 g. The other patient had neurological symptoms 6 weeks after clioquinol was first given, having received 65 g, the average daily dose being 1.5 g, and vision was impaired after 765 g had been administered. On examination 12 and 14 months after clioquinol had been discontinued, the first patient's vision was slightly improved, but he was otherwise unchanged, while the vision of the other patient was unchanged, but she had otherwise deteriorated slightly neurologically. Electrophysiological examinations confirmed the clinical observations. A multifactor etiology of the syndrome: neurotoxicity of clioquinol, paraneoplastic neuropathy and malabsorption, is discussed.


[False diagnosis of space occupying lesions as multiple sclerosis (author's transl)]. Two cases are described in which severe neurological deficits arose because of the wrong diagnosis of Multiple Sclerosis over many years. In both cases the correct diagnosis of a space occupying lesion was possible by clinical signs, particularly by the X-ray examination.


A revised boron-neutron capture therapy for malignant brain tumors. II. Interim clinical result with the patients excluding previous treatments. Fifteen brain tumor patients were treated with slow neutron. It proved to extend life span of terminal glioblastoma patients irresponsive to Co-60, to 2 years, but quality of survival is poor due to complications of previous treatments. Two glioblastoma patients excluding other treatments, the only genuine Boron-neutron capture therapy cases, have been living for 39+ and 34+ months working full-scale without neurological deficit.


[Colour naming deficit in aphasic and non-aphasic brain damaged patients (author's transl)]. We have examined the severity and quality of aphasic misnaming in colour naming tasks as compared to errors of patients with right-sided brain damage and to patients with posterior disconnexion syndrome. The investigation is based on 80 aphasic patients, 20 patients with right-sided brain damage and 80 normal control subjects. Differences in the performance for colour and object naming are discussed. The main results were: 1. The subtypes of aphasia did not differ quantitatively in both types of performance. All aphasics made significantly more errors on colour naming than on object naming tasks. This difference, however, was numerically too small to have practical value. 2. Unlike in the corresponding understanding tasks, both types of naming tasks differentiated between aphasic and non-aphasic patients, the aphasic group performing poorer. 3. The four subtypes of aphasia had similar error scores in the 10 colour tasks. We observed certain regularities in the type of paraphasic misnaming for the total group of aphasics. The subgroup of amnesic aphasia was characterized by a strategy of modifying the colour terms similar to the description of use encountered in the language behavior of these patients. 4. Patients with posterior disconnexion syndrome in general make 7 errors out of 10 tasks in colour naming, in contrast to 2 errors out of 10 tasks in object naming. For aphasic patients the relation is 3.5 to 2.5 errors.


Depth perception and location of brain lesions. Depth perception was examined in 50 patients with brain lesions and in 16 controls using a polaroid test (Titmus). Error percentage and response time were measured. Intellectually impaired patients performed significantly worse than intellectually normal patients. On the other hand, location of the cerebral lesion (right, left, or generalized) had no significant effect; zero error percentages were observed among intellectually normal patients even with right or left parietal lesions. Intellectually normal patients did not differ from healthy controls.


[Cerebellar movement disorders in monkeys. Comparison of rapidly alternating and slower target movements during cooling of the dentate nucleus (author's transl)]. The effects of short reversible cooling of the dentate nucleus in two groups of 3 and 4 cebus monkeys, with two different types of ipsilateral elbow movements, have been studied. One group was trained to turn a moving handle back and forth rapidly between two mechanical stops, while the second group was trained to move the handle between two target zones. Brief blocking of the dentate nucleus caused a delayed termination of contraction of the agonistic muscles (hypermetria) near the mechanical stop for very rapid, ballistic, alternating arm movements and, consequently, delayed initiation of the antagonistic return movement. The resulting increase of the duration of a single movement was not caused by a reduction of the peak acceleration of the movement. For the slower target movements, dentate nucleus cooling caused shortening of agonistic muscular contraction (hypometria) with corresponding, saccadic movement corrections. The frequency of the "movement tremor" lay between 3 and 5 Hz. The average velocity maxima during dentate cooling did not change. The findings indicate that different types of movements exhibit different disturbances of the movement pattern during the period of functional elimination of the same anatomical structure. The results indicate that the dentate nucleus and cerebellar hemispheres take part in preprogramming movement duration (Kornhuber) for rapid ballistic movements. In slower target movements, the dentate nucleus may be involved in sectional preprogramming of step movements.


[Bacillary dysentery with involvement of the central and peripheral nervous system (author's transl)]. A man, age 50, fell ill with polyneuropathy followed by parkinsonism and organic psychosis and died in a shock 6 weeks later. Serologic examination suggested bacillary dysentery, but the patient had no diarrhoea. The neuropathological examination did not reveal any organic substrat of parkinsonism. Peripheral nerves showed mucoid degeneration, segmental demyelination and lymphocytic infiltration of peri- and endoneurium. Many Renaut bodies were found which seemed to arise from mucoid masses organized by cells of the endoneurium. Polyneuropathy and parkinsonism are well known neurological complications of bacillary dysentery and favour this diagnosis in accord with the serological findings.


Movement-associated cortical potentials in unilateral cerebral lesions. Movement-associated cortical potentials (MP) associated with thumb-opposition or fist-clenching were studied in 20 patients with unilateral cerebral lesions and in 8 healthy subjects. MP was abnormal in 16 patients. The most frequent abnormality was an attenuation of the N component or readiness potential, recorded on the central region contralateral to the contracting muscle, on the affected hemisphere. The occurrence and severity of MP abnormalities seem to be influenced by an intensity and extension of the lesion, rather than by how much disability is present in the affected limb. There was no significant correlation between the abnormalities of MP and of EEG. The most useful index of MP for comparing two hemispheres was shown to be the amplitude and wave form of the N component or readiness potential contralateral to the contracting muscle.


DC potentials of temporal lobe seizures in the monkey. In 8 monkeys, made epileptic by alum or penicillin injection into temporal lobe structures, 40 seizures were studied by both DC cortical potential and subcortical EEG recordings. Eighteen seizures of lateral temporal origin had an abrupt negative DC potential shift of 0.5 to 2.0 mV in and around the focus. The frontal, parietal and occipital cortices did not develop DC potential changes, perhaps due to the limited propagation of the neocortical seizures. Twenty-two seizures of medial temporal origin showed a negative shift of the anterior, inferior or lateral temporal cortex in 85% of seizures. The other 15% had a positive or no shift. In hippocampal seizures, a positive displacement was sometimes seen prior to the main negative shift in the lateral temporal cortex. The remote cortex developed only a minimal positive shift in 30% of the mediotemporal seizures. A marked negative shift in the frontocentral cortex was the first sign of impending generalization, which may result from a series of chain reactions with seizure propagation, involving more and more structures of the brain. Registration of DC potentials in temporal lobe seizures may give insight into the nature of abnormal EEG activities and to some extent into the origin of seizures.


Thrombosis and intracranial tumors.

Thrombosis and intracranial tumors. 334 necropsy reports of intracranial neoplasm from an autopsy material over 13 years were reviewed to study the relationship of intracranial tumors to vascular thrombosis. The incidence of venous thrombosis in intracranial tumors was found to be 27.5% while that of a control group without malignancies taken at random from the autopsy material was 17%. The difference gives a statistical significance of P less than or equal to 0.05. The parameters of sex, surgical intervention, the malignancy and the histological type of the tumor apparently dod not affect thrombus formation to a statistically significant degree. There is increased thrombosis frequency with increasing age. The presence of hemiparesis or hemiparalysis does not affect the incidence of thrombosis. However, it determines to a great degree the lateralization of the thrombus.


Duchenne muscular dystrophy and poliomyelitis. A study of dystrophic-denervated muscle. A systematic study of dystrophy-denervation in human muscle showed minimal morphometabolic differences between dystrophic and dystrophic-denervated muscle. The only certain conclusion is that denervation influences the rhythm of evolution of the dystrophy without impressing any of the few characteristics considered at present as peculiar to denervation.


[Spontaneous carotid cavernous fistula in a case of hereditary hemorrhagic telangiectasia (Osler-Rendu) (author's transl)]. A 37-year-old woman suffered from headaches, right facial pain, double vision and occasional tinnitus. On examination there was only a slightly dilated right pupil, weakened corneal reflex and capillary bleeding from telangiectasia of both hands. Similar bleedings occured in her mother (mouth and hands) and son (nose). The arteriography of the right carotid artery showed a cavernous sinus fistula with a small shunt. Steady compression of the carotid artery by hand caused a marked improvement in the subjective symptomatology.


Racial predilection in multiple sclerosis. Comparisons between the geographic distribution of multiple sclerosis and the habitats of various racial groups showed that racial factors alone could not explain the increase in prevalence of the disease with latitude. Racially similar groups living in different areas had different frequencies of multiple sclerosis. Conversely, racially different groups, living in the same area, had similar prevalence rates of multiple sclerosis. Moreover, migrants moving from one environment to another at a young age (before adolescence) appeared "to acquire" the risk of multiple sclerosis of the new environment. These observations suggest than an environmental factor independent of race influenced the risk of acquiring multiple sclerosis. Nonetheless, some genetic factors associated with race may also be implicated, for example, HL-A tissue antigens (perhaps by virtue of a common association with the immune response (Ir) gene), the Gm and Inv immunoglobulin characteristics and skin pigmentary characteristics (perhaps through interactions between pigmentation and calcium metabolism). The specific environmental factors determining risk of multiple sclerosis and the mechanism whereby the racial (genetic) factors may influence risk remain to be elucidated.


The internal auditory artery: (embryology, anatomy, angiography, pathology). A review of the literature on the embryology, anatomy and angiography of the internal auditory artery has shown that there may be considerable variation as to the origin and number of internal auditory arteries. The present study, based on serial magnification angiographies of the internal auditory artery, has demonstrated 7 variants of the origin of this artery although in 45.4% of cases the internal auditory artery arose from the anterior inferior cerebellar artery. For the diagnosis of pathological processes in the cerebellopontine angle (tumors, sudden deafness, vascular abnormalities) magnification angiography is of special importance. Acoustic neurinomas in particular can be diagnosed early on the basis of vascular dislocations and possible tumor staining. Magnification angiography is the method of choice for the demonstration of vascular malformations. The etiology of sudden deafness can not be determined by angiographic studies alone, due to the fact that the internal auditory artery can show considerable variations as regards length and caliber. Magnification angiography of this region should be carried out in all cases with the clinical picture of a cerebellopontine angle lesion in order to achieve diagnostic clarification.


[Measurement of attention deficit in the course of intoxications (author's transl)]. The level and course of attention was measured hourly in 9 drug intoxicated patients after a suicide attempt over periods which varied between 12 and 72 hrs. Attention was measured by the use of two additive 5 step scales for susceptibility to stimulation and reactivity, which were developed by the authors in earlier investigations and proven to be very reliable. Although, the original data set of attention measures was different among the patients, some common features could be elaborated: 1. The level of attention varies very little within 1 hr. Differences greater than one step on the scales were rarely observed between two measurements. 2. The mean course of recovery from attention deficit is linear throughout the scales while the variance is substantial at each step of the scales. For quantification of attention deficit a measure was defined which gives the relation between the actual deficit and full attention. Since the correlation between both scales is high over the whole observation period, it was concluded that the intoxication alters the level but not the structure of attention.


[Myopathia myotonica. A new type of hereditary muscle disease (author's transl)]. A new type of hereditary muscle disease, characterized by weakness and painful spasms during effort, without electrical activity in the shortened muscles, is described. These phenomena are limited principally to the upper limbs. In addition we found electromyographical signs of a generalized myotonic syndrome. The histological and histochemical investigations reveal only minimal non-specific signs of myopathy. The activities of CPK and aldolase in the blood serum are increased at times. A normal elevation of venous lactate was observed during ischemic work. The biochemical studies of muscular tissue exhibit normal activities of the analyzed enzymes, especially as regards phosphorylase. An increased concentration of calcium ions in blood serum may be related to the contraction during strenuous work; it is known that calcium ions are an important factor in the contraction-relaxation cycle of striated muscle. The age of manifestation varied from 4 to 33 years in 4 cases of the relatives observed. The disease shows no signs of aggravation as to the severity and extent of the disorders. The nature of the underlying metabolic defect is still unknown.


[Irritation of the lumbar plexus by an aneurysm of the abdominal aorta (author's transl)]. Case report on a 67-year-old patient presenting sensory disturbances involving the segments L1--L3 and absence of the knee jerk in the left leg. At postmortem examination an arteriosclerotic aneurysm of the aorta abdominalis was found to be the cause for the lesion of the upper lumbar roots.


[The occurrence of diabetes mellitus, diabetes insipidus, optic atrophy and neurogenic deafness in one patient (author's transl)]. Report of a further case with coexisting diabetes mellitus, diabetes insipidus, optic atrophy und neurogenic deafness. It was not possible to find a cause for these symptoms. The cases reported in the literature are brought together and discussed.


3H-thymidine autoradiography of CSF cells in primary reticulum cell sarcoma of the brain. CSF cells in a case of primary reticulum cell sarcoma of the brain with diffuse subarachnoid spreading were examined by 3H-thymidine autoradiography. Immediately after lumbar puncture, the CSF withdrawn was incubated at 37 degrees C for 1 hr with an admixture of 3H-thymidine at a rate of 1 muCi/ml CSF. The cells were collected by centrifugation and the microautoradiographic procedure was performed. The labeling index (L.I.) of the total CSF cells was 10.5%, and when non-neoplastic cells, i.e. polymorphonuclear leukocytes, small lymphocytes, monocytes etc., were excluded, the real L.I. of the tumor cells in the CSF was supposed to be more than 14.4%. Referring to the results of various brain tumors reported in the literature, this belongs to the highest labeling group. The high L.I. of the tumor cells in this case was well consistent with the extremely rapid clinical course. It should be stressed that the examination of CSF cells by 3H-thymidine autoradiography in cases of brain tumors could be one of the valuable methods indicating the DNA synthesis of the tumor cells, which is an important parameter of malignancy.


[The diagnostic value of the F wave latency (author's transl)]. The N. medianus was stimulated electrically in 44 healthy persons and in 30 unselected diabetics. Latencies of the F wave were measured, corrected for different lengths of the upper extremity, and used for computation of nerve conduction velocity. In comparison with conventional motor nerve conduction velocities the F wave latencies showed a more pronounced difference between the normal and the pathological (diabetic) range. The F wave nerve conduction velocities also varied less than conventional nerve conduction velocities. The explanation for this might be that the F wave recordings are the result of a full anti- and orthodromic impulse propagation which travels 5 to 6 times further than the impulse measured with conventional nerve conduction velocity methods. Therefore small delays of nerve conduction will be amplified and can be better recognized by F wave latency measurements. The present findings suggest that F wave latencies may be more sensitive than conventional nerve conduction velocities in evaluating mild cases and in identifying the initial phases of neuropathies.


An electro-oculographic study of ocular bobbing and intermittent vertical oscillations occuring in the same patient. A surviving patient with two rare types of abnormal eye movements, typical ocular bobbing and subsequently developed intermittent vertical oscillations of the eyes, is described, and a follow-up series of electro-oculographic recordings of these eye movements is analyzed. It is suggested that these two types of vertical eye movements might have a common pathophysiological background and the activity of remaining functional oculomotor pathways may play an important role in their occurrence.


The relationship between target, targetoid, and targetoid/core fibers in severe neurogenic muscular atrophy. In the m. tibialis anterior of a 68-year-old man with rapidly developing denervation atrophy in the legs since 1/2 year prior to death from heart stroke, abundant unifocal concentric fiber changes, such as target, targetoid/core, and targetoid fibers could be observed. Besides, large vacuolized fibers with multiple changes resembling cytoplasmic bodies in the peripheral zone were present as well; they are interpreted as fibers with multicentric target or targetoid formations. The target fibers displayed a broad variation of their outer appearance suggesting a continuous transition to targetoid/core fibers (with a dense center) and targetoid fibers (with a central change to aquous sarcoplasm showing a paucity of fibrillar structures). Very few fibers with a central densification of fibrillar material with or without a thin intermediate zone were fairly akin to core fibers of central core disease; others were more alike the type of targetoid fibers, previously described in the literature, showing a dense target-like center; both were summarized under the term, inaugurated by Engel et al. (1966), "targetoid/core fibers". Simultaneous occurrence of the different kinds of concentric fiber changes suggested a strong relation between all of them in the sense of representing different developmental stages of the same pathogenetic process. Thus, the central core disease, for instance, might be a disorder with a generalization of concentric fiber changes having come to arrest in the earliest stage of development.


[X-ray studies of the brain as a basis for stereotaxy (author's transl)]. All attempts to reconstruct the topography of the brain in the living from studies of animal material are handicapped by technical difficulties. The best method is to compare exact X-ray pictures, which have been taken under stereotactic conditions. From a large collection of such X-rays the authors have composed contours of the internal table of the skull and of the ventricles, which best match the brains, selected for the Schaltenbrand-Bailey sterotactic atlas. For practical purposes these contours were combined with the transparent overlays for the nomenclature and the border lines of the different parts of the basal ganglia, which have been used in the myelin sections part of the atlas. A comparison of our sagittal series with the new X-ray findings shows, that the sagittal schemata of the atlas represent an extreme variation in the position of the Meynert axis and of the contours of the 4th ventricle. We have chosen a new axis system for the hindbrain, which corresponds to the average of our brains in constructing a new set of typical overlays for the atlas. The contour of the posterior fossa had to be completed. An independent axis system dor the structures of the 4th ventricle was developed, consisting of the base of the 4th ventricle, and a tangent, to the upper contour of the pons. In sterotactic procedures the axis systems for the forebrain and the hindbrain should be used independently. The results obtained are the basis for a new series of lantern slides which can be projected against the X-ray pictures with the Würzburg stereotactic equipment. In the course of this investigation we discovered a source of error. When air enters the puncture hole of the dura, the brain may sink back, so with the patient lying on his back, all structures may shift a few millimeters towards the occipital region. When the patient is lying on his side, as during an approach to the amygdala through the planum temporale, the ventricular system may collapse, so that almost no air is visible in the ventricles and the 3rd ventricle may appear to be in the lower hemisphere, the dislocation being more than 5-8 mm. But filling the ventricle with air through the ventricular catheter is sufficient to blow up the brain and to restore the normal topography.


[Torpid degeneration of the lateral pallidum with Bielschowsky bodies. Light and electronmicroscopic findings (author's transl)]. In a 60-year man with a very slowly progressive motor disturbance probably of torsion systonic or choreoathetotic type which started in childhood, we found an isolated degeneration of the exterior pallidum with intraneuronal deposition of Bielschowsky bodies and only slight deficit of neurons. The Bielschowsky bodies show the same structure as corpora amylacea and Lafora bodies. They might also be the result of a cellular disturbance mainly concerning the carbohydrate metabolism. In this case they are localised only in the neurons of the exterior part of the pallidum--in contrast to the organogenitically otherwise deriving (from the nucl. entopeduncularis) intact neurons of the pallidum internum--and they induce, after an excessive accumulation of the metabolites, not till extremely late loss of neurons. This circumstance explains the very torpid progress of this disease.


[The problem of early diagnosis of brain tumours causing seizures only (author's transl)]. The diagnosis of brain tumour could not be made in 91 cases at the first investigation in a group of 1155 brain tumours. Slowly growing gliomas causing only epileptic fits and no other symptoms are especially difficult to diagnose. Of 21 personal observations of tumour seizures, in which the diagnosis of the neoplasm was missed at the first investigation in hospital, 9 were oligodendrogliomas, 5 astrocytomas, 3 glioblastomas, 2 spongioblastomas, 1 gangliocytoma and 1 a metastasis. They were all located in the frontal or centroparietal region. In most cases the seizures appeared during the third or fourth decade. The average interval between the first epileptic fit and the tumour diagnosis was 8.2 years in cases of oligodendrogliomas and 2.2 years in astrocytomas. 5 patients had major seizures, 2 had psychomotor attacks and all the others suffered from partial epilepsy. Anticonvulsive therapy was often successfull; either the frequency of the fits diminished or, in 2 cases, the character of the seizures changed. 18 patients had a normal neurostatus at time of the first investigation. Only 3 patients had a slight difference of physiological reflexes, but no other pathological signs. In none of the patients did investigation of the CSF, skull X-rays, brain scanning, pneumencephalography or cerebral angiography first lead to the diagnosis of a brain tumour. The EEG alone showed focal signs corresponding to the location of the tumour in about 50% of the cases.


[To the differential diagnosis of cranial nerve lesions: the progressive necrotising external otitis (author's transl)]. A review of necrotising external otitis, a relatively unknown and dangerous disease, brings out that, initially, it has three characteristics: a granulating necrotising ostitis of the external meatus, extreme pain and a yellowish green secretion. It is always caused by a pseudomonas infection and in almost all cases the patients suffer from diabetes mellitus. If the condition is not recognized in good time and an extensive debridement of the bone involved not performed promptly, ostomyelitis of the base of the skull may follow with involvement of cranial nerves. Severe chronic osteomyelitis of cervical vertebrae occurred in one of our cases. The neurologist must bear this disease in mind in the differential diagnosis when cranial nerves are affected because the nerve disturbances may become evident only after the local condition has subsided or the nerve deficits may be more prominent than and obscure the local ear condition. The most commonly involved nerve is the facial although there may be multiple cranial nerves involved including the third through the twelfth. If the cervical vertebrae become affected there may be nerve root lesions. A torpid meningoencephalitis may also occur. Close cooperation between otologists and neurologists is necessary to recognize and treat these conditions properly.


Preparation and chemical characterization of radioiodinated bleomycin. None of the radionuclides with which bleomycin has been labeled have chemical and nuclear properties that are entirely satisfactory for in vivo tumor localization. Bleomycin has been radioiodinated by the iodine monochloride, chloramine-T, and lactoperoxidase methods. Iodine monochloride proved to be the preferred method and conditions were developed whereby 80% of radioiodide was covalently bound to bleomycin. Bleomycin (140 mug) was added to 200 mul of saline/citrate buffer (pH 7.0) followed by radioiodide and iodine monochloride. This reaction mixture was incubated for 1 hr and purified by Sephadex G-10 chromatography. The iodine monochloride reaction product underwent hydrolytic deiodination in vitro at a rate of about 1.2%/day (0.15 M NaCl, 37 degrees C). Bleomycin A and B components were radioiodinated with equal efficiency on a mole fraction basis.


Cobalt-labeled bleomycin--a new radiopharmaceutical for tumor localization. A comparative clinical evaluation with gallium citrate. The efficacy of 57Co-labeled bleomycin as a tumor-scanning agent was evaluated in 50 patients with malignant tumor. In terms of sensitivity, labeled bleomycin was found to be a superior agent for malignant tumor detection when compared with 67Ga-citrate (p less than or equal to 0.005). Maximum sensitivity (96%) was found in epidermoid carcinomas. Because of rapid blood clearance and rapid excretion by the kidneys, interpretation of 57Co-bleomycin scans is facilitated.


Effects of nutritional factors on the development of ethanol-induced fatty liver in KK and KK-Ay mice. KK and KK-Ay mice developed a steady and reproducible fatty liver when they were given free access to an ethanol solution as a drinking fluid for 10 to 20 days. The present studies were undertaken to elucidate effects of nutritional factors on liver fat contents of the mice given water or ethanol solution. In contrast to cornstarch, sucrose tended to increase the liver fat of control mice. A higher concentration of dietary casein lowered the liver fat of control mice, whereas the dietary concentration of cottonseed oil did not significantly affect the liver rat levels either in control or ethanol groups. Thus the standard basal diets favorable for the development of the alcoholic fatty liver have been established, for example, 10% cottonseed oil, 25 and 30% casein, 58.4 and 53.4% cornstarch for KK (12-15 weeds old) and KK-Ay (5-10 weeks old), respectively. Neither choline, myoinositol, nor any lipotropic agent tested prevented the development of the alcoholic fatty liver. Unlike in rats, orotic acid did not induce a fatty liver but rather alleviated the ethanol-induced fatty liver in these mice.


An alcian blue-phloxine method for the gross demonstration of squamous metaplasia in the larynx. A macroscopic method using the dyes Alcian blue and phloxine is described for the investigation of squamous metaplasia in the larynx. The method also demonstrates an intermediate type epithelium about which little is known. The gross staining of the tissues is based upon the differential penetration of the dyes through squamous, intermediate and respiratory epithelia, and on the strong chemical complex formed between Alcian blue and mucin. The method is particularly useful for research purposes owing to the permanency of the staining result.


Ultrastructure of the invasion of Eimeria magna sporozoites into cultured cells. Monolayers of bovine kidney cells were overlaid with Eimeria magna sporozoites and observed with phase-contrast optics until penetration of the cells by the parasites had begun.  Cells and penetrating parasites were fixed with glutaraldehyde and OsO4-containing ruthenium red, dehydrated, and embedded in situ.  Cells being penetrated were selected for study in the electron microscope.  The lack of intracellular staining with ruthenium red and intact plasmalemmas of cells being penetrated, was accepted as evidence that the sporozoites did not disrupt the plasma membranes.  The sporozoite caused invagination of the host cell plasmalemma until the parasite was entirely within the cell, after which the invagination was sealed off by short pseudopodia enclosing the sporozoite within a membrane=lined vacuole inside the cell.  Often myelin-forms, apparently of host cell origin, were seen in the space between the sporozoite and the cell.


Effects of bleomycin on mouse bone-marrow stem cells. The mouse hematopoietic stem-cell population was tested by the spleen colony technique for effects of the antineoplastic agent bleomycin (BLM). The time response of normal bone marrow was investigated by a single dose of BLM (400 mg/kg) between 0 and 72 hours. The dose response was studied over a wide range of doses (from 40 to 1,600 mg/kg) at a 4-hour exposure. Additional experiments concerned 1) the fraction of colony-forming units in the S phase after BLM administration (by means of pulse hydroxyurea treatment), 2) the response of bone marrow stimulated by endotoxin, and 3) the effects of split-dose treatments. The relatively low toxicity of BLM on both the differentiated and stem-cell populations of unstimulated bone marrow was confirmed and detailed. This drug exhibited peculiar, proliferation-dependent cell inactivation kinetics. Furthermore, BLM induced parasynchronous behavior in the unstimulated stem-cell population. The various aspects of BLM action are discussed with regard to its use in cancer chemotherapy.


Detection of a TL(+) murine leukemia cell line that resists the cytotoxic effects of guinea pig complement and specific antiserum. RADA-1 cells [H-2a thy-1b; thymus-leukemia (TL) 1, 2, 3], a radiation-induced murine leukemia cell line maintained by serial transfer in histocompatible recipients, resisted lysis by guinea pig complement (GPC) and TL 1, 3; TL 2; or TL 1, 2, 3 antiserum. The cells expressed TL antigenic specificities as determined by indirect fluorescent antibody methods, the direct isolation of TL antigens from the cells, and the capacity of the cells to reduce known titers of TL antisera. GPC was consumed to the same extent during the reaction of resistant cells and TL antisera as occurred in the reaction of sensitive cells (killed under similar conditions) and TL antisera. RADA-1 cells were not nonspecifically resistant to complement (C)-mediated lysis; they were killed in the presence of H-2a antiserum and GPC. The TL antisera contained antibodies for TL determinants. They stimulated the C-mediated lysis of ASL-1 cells (TL 1, 2, 3) and thymocytes from strain A mice (TL 1, 2, 3). The TL antigens of resistant RADA-1 cells underwent antigenic modulation, the reversible disappearance of TL antigens from the cells stimulated by specific antiserum. After the cells were treated with neuraminidase, they became susceptible to the cytotoxic effects of aliquots of the same TL antisera and GPC used previously.


Complement sensitivity of somatic hybrids of a complement-resistant murine leukemia cell line. RADA-1 cells (H-2a), a murine leukemia cell line maintained by serial transfer in histocompatible recipients expressing thymus-leukemia (TL) 1, 2, 3 antigenic determinants, resisted the cytotoxic effects of guinea pig complement (GPC) and TL antiserum. The cells expressed a lower density of TL antigens than did ASL-1 cells, another TL(+) leukemia cell line expressing the same determinants and susceptible to complement (C)-mediated lysis. Stable somatic cell hybrids of RADA-1 cells and LM(TK)- cells (H2k), a TL(minus) thymidine kinase-deficient mutant of mouse L cells, were selected in hypoxanthine-aminopterin-thymidine medium. The hybrid cells expressed the H-2 antigens of both parents and shared a hybrid karyotype. They formed TL 1, 2, 3 antigens as determined by immunofluorescence, mixed hemagglutination methods, the direct isolation of TL antigens from Nonidet P40 extracts of the cells, and the capacity of the cells to reduce by absorption the known titers of TL antiserum. These hybrid cells lost the capacity to resist lysis by TL antiserum and GPC. They were susceptible to the cytotoxic effects of TL 1, 2, 3; TL 2; OR TL 1, 3 antiserum and GPC, even though the density of TL antigens associated with the cells was approximately 25% of their resistant RADA-1 parental cells. These results indicated that the mechanism of resistance to C-mediated lysis was genetically separable from the expression of TL antigens by the cells and that the susceptibility of the cells to the cytotoxic effects of antiserum was related only in part to the density of TL antigens expressed by the cells.


Inhibition of purified DNA polymerase of RNA tumor viruses by fluoranthene derivatives and analogues of tilorone hydrochloride. At concentrations of 7 times 10(-6) to 7 times 10(-5) M, derivatives consisting of the polycylic ring structures fluoranthene, fluorenone, fluorene, anthraquinone, xanthenone, and dibenzofuran with appropriate amine side chains inhibited by over 90% the purified RNA-directed DNA polymerase of avian myeloblastosis virus acting on poly(deoxyadenylate-deoxythymidylate) [poly(dA-dT)]. Of these, only the fluoranthene derivatives were strong inhibitors of the viral DNA polymerase directed by polyadenylate-oligodeoxythymidylate [poly(A)-(dT)12-18]. Low levels of fluoranthene derivatives (1 times 10(-5) M) also strongly inhibited polymerase with polyinosinate-oligodeoxycytidylate [poly(I)-(dC)12-18], activated calf thymus DNA, and viral 70S RNA as templates, but not with polycytidylate-oligodeoxyguanylate as template. A comparison of the activity of 11 fluoranthene derivatives with different side chains showed that the structure of the amine side chain influenced both the extent of antipolymerase activity with a given template and the relative inhibition with different synthetic DNA and RNA templates. The naturally occurring polyamines, spermine, spermidine, and putrescine, did not inhibit the activity of the viral DNA polymerase. Studies on the mechanism of action indicated that the synthetic derivatives inhibited polymerase activity by binding to the template and not to the enzyme: 1) inhibition by fluoranthene derivatives was overcome by the addition of excess template including poly(dA-dT), poly(A)-(dT)12-18, poly(I)-(dC)12-18, viral 70S RNA, and activated calf thymus DNA; 2) the degree of inhibition by fluoranthene derivatives was unaffected by the addition of the creased viral DNA polymerase; 3) with the same template, Escherichia coli DNA-directed RNA polymerase and the viral RNA-directed DNA polymerase were inhibited to about the same extent; and 4) the derivatives formed a complex with DNA, poly(I), and poly(A) that was stable to exclusion chromatography on Sephadex G-100. Several derivatives also had biologic activity, since they blocked the ability of the murine sarcoma virus to transform cells.


Particle-associated RNA dependent DNA polymerase and high-molecular-weight RNA in a human cell line derived from polycythemia vera bone marrow. A particle fraction with a density of 1.15-1.19 g/cm3 was isolated from the cytoplasm of a human cell line established in culture from the bone marrow of an untreated patient with polycythemia vera. Electron micrographs of cross sections of cells and cell homogenates revealed virus-like particles on which DNA could be synthesized. An RNA-dependent DNA polymerase, isolated from the particles, preferred poly(rA)-oligo(dT) over poly(dA)-oligo(dT) and was able to polymerize deoxyguanosine monophosphate in a reaction stimulated by poly(rC)-oligo(dG).


Immunoprophylaxis and cytotoxic effector cells against EL 4 leukemia induced in syngeneic C57BL/6J mice by use of irradiated EL 4 cells. Tumor-specific immunoprophylaxis was achieved in C57BL/6J mice against EL 4 leukosis cell challenge by sensitization of the syngeneic host with multiple ip injections of irradiated EL 4 cells. A minimal radiation dose was used to replication-block EL 4 cells before inoculation, as defined by dose-response analysis of irradiated EL 4 cells. Multiple ip injections of irradiated EL 4 cells stimulated development of significant, yet relatively low, levels of cytotoxic lymphoid activity (CLA) in lymphoid cells of the peritoneal exudate as measured by in vitro 51Cr-release cytotoxicity assays. The specific temporal and frequency dependencies of the inoculation regimen for achieving immunoprophylaxis indicated that, in addition to CLA, other, short-lived, immune processes were important in the tumor rejection. These observations showed the capacity of the C57BL/6J host for tumor-specific immune recognition and rejection of the syngeneic EL 4 leukemia. The tumor rejection could be elicited solely by inoculations of irradiated EL 4 cells and did not require exogenous amplifiers, such as immunoadjuvants, chemical modifiers, and/or allogeneic immune information transfer.


Alteration of cell-surface antigenicity of the mouse plasmacytoma. I. Immunologic characterization of surface antigens masked during successive transplantations. Alterations of membrane antigenicity of IgA-synthesizing plasmacytoma cells (58-8) induced in a BALB/c mouse were investigated with rabbit antisera against 58-8 transplanted from 7-8 generations (anti-58-8) and mouse antisera against H-2d (anti-H-2d). The 58-8 cells transplanted (TP) for 8 generations (TP8), showed moderate susceptibilities to anti-58-8 [cytotoxicity index (CI) = 72%] and to anti-H-2d (Cl = 30-40%). At TP12, the susceptibility to anti-58-8 remained (tCl approximately 50%) but there was none to anti-H-2d (Cl = 0-10%). After TP13, 58-8 cells had detectable reactivity neither with anti-58-8 nor with anti-H-2d before proteolysis. However, antigenicity was demonstrated after treatment of cells with proteases: Anti-58-8 and anti-H-2d became cytotoxic to pronase-treated 58-8 (P-58-8) and these cytotoxic reactivities of antisera could be completely absorbed with P-58-8- but not with "intact" 58-8. Anti-H-2d absorbed with P-58-8 had no cytotoxicity to BALB/c spleen cells. Absorption of anti-H-2d with BALB/c spleen cells did not show any measurable cytotoxicity to P-58-8. Appearance of antigenicity after proteolysis was transient and reversible. When P-58-8 was incubated in vitro, susceptibilities to antisera were decreased. Actinomycin D and puromycin inhibited the loss of susceptibility; these metabolic inhibitors suppressed the "masking" of antigenic determinants with protein-like material(s). In early generations (TP8), 58-8 cells were susceptible to antiserum to bone marrow-derived lymphocytes (anti-B) and complement. The cells completely absorbed the cytotoxicity of anti-B against spleen cells. After additional transplantations (TP14, TP30), susceptibilities to anti-B were no longer detectable and no removal of the cytotoxicity was shown, though the cells were pretreated with pronase. Anti-B failed to kill MOPC-31C.


Antitumor effects of antibody-diphtheria toxin conjugates. III. Cyclophosphamide-induced immune unresponsiveness to conjugates. Of 9 hamsters immunized with diphtheria toxin-hamster gamma-globulin conjugates and rechallenged with this antigen 15 days later, 8 exhibited neutralizing antibodies against the conjugates 22 days after the initial immunization. Hamsters identically immunized, but also given cyclophosphamide (CY) 20 hours after the first immunization, had a much lower incidence of positive responses (2/10) and lower antibody levels in the positive responders. After further immunization with the conjugates, all CY-treated hamsters eventually developed neutralizing antibodies, but exhibited a reduction in antibody levels after a second exposure to CY. Hamsters immunized with diphtheria toxoid developed antibodies that were effective in neutralizing diphtheria toxin, but less effective against the diphtheria toxin conjugates.


Production of antibody to individual polypeptides derived from purified hepatitis B surface antigen. Purified preparations of hepatitis B surface antigen (HBsAg) were solubilized with sodium dodecyl sulfate and urea under reducing conditions and subsequently fractionated by preparative sodium dodecyl sulfate-urea polyacrylamide gel electrophoresis (PAGE). Pools of the individual fractions eluted from the preparative PAGE were concentrated and purified further by analytical PAGE. Five purified polypeptides were isolated from HBsAg, types adw and ayw, with molecular weights of 19,000, 24,000, 27,000, 35,000, and 40,000. Each preparations was emulsified in Freund complete adjuvant and injected into guinea pigs. Antibody to each HBsAg type was measured by radioimmunoassay. The 19,000 molecular weight polypeptide derived from ayw particles and the 27,000 molecular weight subunit obtained from both types failed to elicit an antibody response. The other three polypeptides derived from the ayw particles elicited group-specific antibody responses. Similar group-specific reactivities were observed in the testing of anti-adw 35,000 and anti-adw 40,000 molecular weight polypeptide sera. However, guinea pigs immunized with the 19,000 and the 24,000 molecular weight polypeptides of the adw type produced antibody that reacted preferentially with adw particles. This indicates that either these subunits carry predominately d determinants or that, because of the low levels of material used for inoculation, no immune response or an undetectable one was elicited to the a or w components.


Quantitation of RNA tumor viruses by spectroscopy of density gradient gels. We have developed a system for virus particle quantitation based on the measurement of the optical absorbance of stained viruses which first have been banded at their buoyant density in an equilibrum 24 to 53% (wt/wt) sucrose density gradient, then fixed in position in the gradient by photopolymerizing an acrylamide-riboflavin mixture in the sucrose, and finally stained and destained. Using plasma from mice infected with leukemia virus (Rauscher) or chickens infected with avian myeloblastosis virus (BAI strain) or suitable controls, we have shown that this technique specifically detects RNA tumor viruses. By using virus stock solutions for which the absolute concentrations were determined by laser beat frequency spectroscopy, we have calibrated the absorbance of the viral bands in terms of virus particle concentration. Using 0.8-ml gradients gels (4 by 45 mm) we can detect as low as 2 x 10(7) viral particles with Coomassie blue staining and 6 x 10(6) viral particles with a more sensitive staining procedure using amido black.


Interferon-directed inhibition of chronic murine leukemia virus production in cell cultures: lack of effect on intracellular viral markers. Extracellular murine leukemia virus (MLV) reverse transcriptase activity was decreased by interferon treatment in four interferon-sensitive mouse cell lines which were chronic MLV producers. In three cell lines which were relatively insensitive to interferon, extracellular enzyme activity remained unchanged by interferon treatment. The concentrations of interferon used had no effect on DNA synthesis or cell replication of AKR,C+ cells which were chronic producers of AKR-MLV. In AKR,C+ cultures interferon treatment also had no effect on the level of intracellular viral reverse transcriptase activity in spite of an inhibition of extracellular enzyme activity. Treatment of AKRC+ cultures with interferon for 9 days inhibited extracellular viral reverse transcriptase levels throughout the period of treatment; however, the intracellular enzyme activity remained unchanged, and concentrations of viral p30 (gs) antigen were increased in the interferon-treated cells. When the cells were washed to remove interferon, however, virus production rapidly rose and intracellular p30 antigen fell to the levels of untreated AKR,C+ cells. These and previously reported results suggested that in interferon-treated AKR,C+ cells virus production is inhibited at a late step in the MLV replication cycle, either directly or through the inhibition of the production of a protein required for virus assembly.


Combined intra-arterial actinomycin D and radiation therapy for surgically unresectable hypernephroma. One primary goal of preoperative radiotherapy for hypernephroma is to reduce the volume of tumor and, therefore, improve the possibility of resection. It is important that this goal be accomplished promptly so that 4 to 6 weeks after radiation therapy nephrectomy can be attempted. A longer waiting period may allow fibrosis of the normal surrounding tissues and make surgery more difficult. In addition, longer waiting periods could theoretically increase the probability of metastasis. Therefore, we plan to continue clinical investigation on the use of combined intra-arterial actinomycin D and radiotherapy as a possible useful means of improving the possibility of prompt surgical resection, since theoretically this regimen may be a method of increasing the effective radiation dose to the hypernephroma without increasing the effective radiation dose to surrounding normal tissue, such as bowel. The method may also have merit as an improved means of palliating selected patients with metastases who are symptomatic from a bulky primary hypernephroma.


Cell-mediated and humoral immunity in mice: cross reaction between lysozyme and S-carboxymethylated lysozyme studied by a modified footpad test. The mouse sensitized by subcutaneous (sc) injection of lysozyme in emulsion of Freund's complete adjuvant (FCA) was shown by a modified footpad test to develop three kinds of hypersensitivities. Injecting lysozyme in 2.5-mul emulsion of Freund's incomplete adjuvant (FIA) into the footpad elicited strong footpad swelling in 30 min (anaphylactic reaction), in 3 hr (Arthus-type reaction) and in 24 hr (delayed-type hypersensitivity; DTH). The mice showing anaphylactic reaction in the footpad test manifested severe active systemic anaphylaxis, and the sera of these animals showed high IgG1 antibody titers with only sparingly detectable or no IgE antibody titers. In the sensitizing system with the use of FCA, the antigenicity of S-carboxymethylated lysozyme (CM-lysozyme) devoid of the three-dimensional conformation of lysozyme was compared with that of the native molecule. CM-lysozyme and lysozyme completely cross-reacted to each other in DTH, but not at all in the anaphylactic or Arthus-type reaction or in IgG1 antibody production. CM-lysozyme was shown also to have the ability to bestow immunological memory for the induction of humoral immunity against lysozyme; intravenous (iv) injection of lysozyme in saline or sc injection of CM-lysozyme-FCA alone failed to induce immediate hypersensitivities and IgG1 antibody production against lysozyme, but pre-sensitization by sc injection of CM-lysozyme-FCA enabled the animal to induce these responses to significant levels when iv injection of lysozyme in saline was given as a booster.


Immunochemical characterization of cell wall protein antigen purified from the cell wall autolysate of Clostridium botulinum type A. The cell wall protein antigen was solubilized from the isolated cell walls of Clostridium botulinum type A by autolysis and purified by diethylaminoethyl-cellulose column chromatography followed by gel filtration on Sephadex G-150. The two fractions showed a high degree of the serological activity and produced a main fused precipitin line in immunodiffusion tests against the homologous antiserum. The fact that antigenic fractions contained various kinds of amino acids but no detectable amounts of amino sugars or carbohydrates suggests that the antigens were principally composed of proteins. The protein antigen possessed multiple antigenic components in immunoelectrophoresis. As serological activity, the antigen was heat-stable and resistant to tryptic digestion but sensitive to the actions of pronase, nagarse or pepsin. The protein antigen appeared to be responsible for the common antigenicity among the proteolytic strains of C. botulinum.


Isolation and electrophoretic study on Mallory bodies from the livers of alcoholic cirrhosis. Mallory bodies (MBs) were isolated from the livers of eight alcoholic patients. The method consisted of homogenization, velocity sedimentation in 60 per cent sucrose solution and two-phase polymer centrifugation using a polyethylene glycol-dextran system. MBs were isolated in large quantity and high purity (more than 95 per cent in some cases). Isolated MBs maintained the characteristic filamentous structures. They were solubilized in 6 M guanidine hydrochloride in 10 per cent or 1 per cent acetic acid by sonication or in 1 per cent sodium dodecyl sulfate with 5 per cent 2-mercaptoethanol and 8 M urea. The solubilized MB protein produced three peaks on Sephadex G-100 chromatography and at least five intense protein bands on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The fractions from control livers contained several bands which were identical with the major components of isolated MB protein. These findings suggest that MBs are not homogeneous protein, but that they are probably complexes of various proteins, and that some components of MBs are present in normal hepatocytes. The exact nature of MBs has yet to be established.


Human glomerular cells in tissue culture. Cells from human glomeruli explanted in tissue culture were grown and subcultivated up to 12 to 13 times. Light and electron microscopic studies revealed these cells to be morphologically distinct from fibroblasts. By electron microscopy, an extracellular material resembling basal lamina was seen and prominent intracellular microfilaments were evident. Immunofluorescent microscopy demonstrated reactivity of heterologous antiglomerular basement membrane antibody with aggregates of extracellular material. Absorption experiments using antiglomerular basement membrane antibody showed that the extracellular materiial shared some antigenic components with glomerular basement membrane. Antibody to cultured glomerular cells stained the mesangium and glomerular basement membrane of normal human kidney. This antibody was nephrotoxic in monkeys, induced proteinuria with proliferative glomerulonephritis, and localized to the mesangium and glomerular basement membrane of monkey glomeruli. These findings and the presence of prominent intracellular microfilaments (contractile elements) suggest that the glomerular cells may be of mesangial origin.


Association and linkage between alcoholism and eleven serological markers. Sera from alcoholics and their nonalcoholic first-degree relatives were analyzed for 11 serological markers. A lower prevalence of the S antigen in the nonalcoholic relatives and linkage in repulsion between the D gene of the Rh system and alcoholism were found. All the alcoholics and the relatives had an SS phenotype, when analyzed for complement C3, compared with a prevalence rate of about 50% in other studies. The first two associations involve blood groups, the latter a serum protein.


Intra-abdominal, angiofollicular lymph node hyperplasia (plasma-cell variant) with an antierythropoietic factor. An 11-year-old girl presented with a refractory hypochromic microcytic anemia, hypoferremia, normoblastic hyperplastic bone marrow, hypergammaglobulinemia, and growth retardation. Many varied treatments failed to produce any improvement. Ferrokinetic studies revealed rapid plasma clearance and increased plasma iron turnover, but impaired incorporation of 59Fe. Excretion of 57Co after an oral dose indicated an increased iron absorption. A (99M)Tc-sulfur colloid scintigram of the abdomen failed to demonstrate abnormal uptake. A nodal mass showing the plasma-cell variant of angiofollicular hyperplasia was removed from the gastrolienal ligament. Follow-up studies at 3 and 6 months revealed complete correction of the anemia, a 4.8-cm increase in height, and normal serum gamma-globulin levels. Serum obtained before operation inhibited the incorporation of 59Fe that was induced by a standard dose of erthyropoietin in the exhypoxic mouse system, and this inhibition persisted in serum obtained 3 days after surgery but disappeared by 6 days. The data suggest that the hyperplastic angiofollicular lymph node (plasma-cell variant) secreted a substance the inhibited erythropoiesis.


Surgery of the thyroid.

Surgery of the thyroid. The sophistication of current surgical treatment of thyroid disorders reflects a better understanding of the pathophysiology. The development of more methods of therapy with antithyroid drugs, beta-adrenergic blocking agents, thyroid hormones, and radioactive isotopes as well as external irradiation has had an important effect on the role that surgery plays in the management of diseases of the thyroid. Iodine, which was use for many disorders, is now probably contraindicated except for preparation of the hyperthyroid patient for surgical treatment. Surgery has now taken its place as one form of therapy for an endocrine organ that is subject to a variety of diseases. It is necessary for the surgeon who undertakes operations on the thyroid to have an understanding of thyroid physiology as well as its pathophysiology.


Comparative trial of amoxycillin and chloramphenicol in treatment of typhoid fever in adults. A randomised clinical trial in 124 adult patients with typhoid fever, proved by blood culture, showed that amoxycillin in a dosage of 1 g. six-hourly for fourteen days is an alternative to chloramphenicol, which has hitherto been regarded as the drug of choice.