Isolation of mammalian cell mutants deficient in glucose 6-phosphate dehydrogenase by means of a replica-plating technique. A replica-plating technique is described which was used for the isolation of G-6-PD-deficient mutants in cultures of mutagen-treated Chinese hamster cells. Mutants were recognized by their failure to stain in a histochemical G-6-PD-specific staining reaction. Four mutants were isolated and characterized by growth properties, stability of their variant phenotypes, and reduced G-6-PD activity. One of these mutants on electrophoresis exhibited a variant G-6-PD and thus is very likely the result of a mutation in the structural gene for G-6-PD.


C and Q bands in long arm of Y chromosomes; are they identical? A phenotypically normal male has a small Y chromosome with no Yq fluorescence, but displays constitutive heterochromatin on the end of Yq. C and Q bands on Yq therefore need not be necessarily identical.


Prenatal diagnosis of congenital anomalies in an intrauterine growth retarded fetus. Chromosome analysis of amniotic fluid cells and amniotic fluid alpha-fetoprotein determinations were used to investigate a fetus with severe intrauterine growth retardation in the third trimester. The karyotype was 47,XY,18+ and increased alpha-fetoprotein levels indicated the presence of congenital malformations. We suggest that when severe fetal growth retardation is detected early in the antepartum course, amniotic fluid alpha-fetoprotein and amniotic fluid cell chromosome studies be done to determine if congenital anomalies may be an etiological factor.


CT banding of human chromosomes. The role of cations in the alkaline pretreatment. This paper describes a study of the role of certain cations in the alkaline pretreatment step used in the CT technique for chromosome band formation. Treatment of human chromosomes with ammonium bases or with the hydroxides of the monovalent alkali metals Na, K, or Li resulted in their rapid disintegration, unless very short treatment periods or diluted solutions were used. In the latter cases a subsequent staining produced a weak G-banding pattern. The chromosomes appeared to be much less sensitive to treatment with the hydroxides of the divalent alkaline earth metals Ba, Sr, Ca, and Mg. Staining after exposure to these hydroxides yielded R-banding patterns. The reduced alkali sensitivity of the chromosomes and the reverse banding pattern formation observed are probably the result of a chromatin stabilization by the divalent cations of the alkaline earth metals. It is proposed that not only in the R-band formation with the hydroxides of the alkaline earth metals but also in that obtained by other techniques, chromosome stabilization plays an important role.


Early detection of pregnancy-associated serum protein using antiserum to placental antigens. Antisera against human placental proteins were developed in goats and rabbits, using immunoadjuvants and a prolonged injection schedule. The antisera were absorbed with normal serum proteins and then tested in immunodiffusion against normal and pregnancy sera. Two bands of precipitation due to pregnancy antigens were observed in pregnancy sera as early as 18 days after conception. Detection of these antigens has possibilities for application as an early pregnancy test.


The use of 3H serotonin release from mast cells of the mouse as an assay for mediator liberation. The release of 3H 5-HT from murine mast cells is shown to be a simple reproducible method for studying the activation of such cells by various agents. 3H-serotonin was taken up by peritoneal cell suspensions in vitro and was released by antigen, Concanavalin A, Forssman antiserum, anti-mouse immunoglobulin, or a polypeptide antibiotic, Cinnamycin.


Antigenic relationships on the diphtheria toxin molecule: antitoxin versus antitoxoid. We used the mouse to produce antisera to native diphtheria toxin and diphtheria toxoid. With these antisera it was possible to distinguish between toxin and toxoid. By gel diffusion analysis, antitoxin detected antigenic determinants on toxin which were not available on toxoid, indicating that some determinants had been lost or altered by formalin treatment. Antitoxoid, on the other hand, showed reactions of identity between toxin and toxoid in gel diffusion. The toxin neutralization titers measured in tissue culture were the same for both antisera. Only antitoxin neutralized the adenosine 5'-diphosphate ribosyl-transferase activity of fragment A, but suprisingly both antisera had significant anti-fragment A titers when tested by passive hemagglutination. It is suggested that some of the anti-fragment A activity in antitoxin affects the enzyme active site, whereas that in antitoxoid does not, implying the existence of a least two independent antigenic regions on fragment A.


Chemical and immunological properties of the type f polysaccharide antigen of Streptococcus mutans. The type-specific cell wall polysaccharide antigen was extracted, purified, and characterized from type f Streptococcus mutans strain OMZ175 and MT557. The antigen was extracted from lyophilized cells with 5% trichloroacetic acid at 85 C for 15 min or saline at 120 C for 30 min. The trichloroacetic acid antigen was chromatographically separated into three antigenic fractions on a diethylaminoethyl-Sephadex A-25 column. Antigen 1 (Ag1P), which was specific for type f antiserum, was further purified by passing through carboxymethyl-Sephadex C-25 and Sephadex G-200 columns. It was a polysaccharide composed of 49% rhamnose and 47% glucose. No reaction was obtained with anti-polyglycerophosphate (PGP) serum. Antigen 2 was reactive with both type f and PGP antisera and contained significant amounts of protein and phosphorus. Antigen 3 was reactive only with PGP antiserum and had no type specificity. The polysaccharide antigen gave a single precipitin band against type-specific antiserum on immunodiffusion and immunoelectrophoresis. The presence of alpha-1,6-glucosidic linkages was indicated by a 90% inhibition of the precipitin reaction by isomaltose and alpha-methyl-D-glucopyranoside, adsorption to and release from a concanavalin A-Sepharose column, and reaction with an S. mutans (type e) glucan antiserum. This antiserum was used to show that the type f polysaccharide antigen did not contain free glucan. An analysis of the antigen released from the antigen-glucan antiserum complex showed the presence of rhamnose and glucose. This released antigen also reacted with an f antiserum, which did not react with commercial dextran. The results show that the type f polysaccharide antigen is the first of those S. mutans type-specific polysaccharides identified to be immunologically related to an S. mutans glucan.


Identification of the stumptailed macaque virus as a new papovavirus. A new viral agent, stumptailed macaque virus (STMV), isolated from uninoculated stumptailed macaque kidney cultures was identified. The virions had the size and morphology of papovaviruses of the simian virus 40 (SV40)-polyoma subgroup, but many of them appeared to have an additional outer envelope. The deoxyribonucleic acid of STMV was a superhelical circular molecule, with a mean length 91% of that of SV40. The antigenic relationship of this virus with other members of the group was examined by immune electron microscopy of isolated virions and by immunofluorescent staining of virus-infected cells. STMV was immunologically distinct from SV40, BK virus (BKV), polyoma virus, and JC virus. Its tumor antigen may be related to those of SV40 and BKV.


Purification and immunochemical characterization of type e polysaccharide antigen of Streptococcus mutans. The type-specific antigen of Streptococcus mutans strain MT703, serotype e, has been chromatographically purified and characterized. Two chromatographic fractions were obtained from saline extracts which reacted with both anti-MT703 whole-cell serum and Lancefield group E serum. The major fraction (eI) was identified as a polysaccharide composed of 37% glucose, 56% rhamnose, 5% protein, and 0.3% phosphorus, whereas the minor fraction (eII) contained 66% protein in addition to 10% glucose and 17% rhamnose. The immunological specificity of these antigens was found to be the same by immunodiffusion in agar gel. Another fraction with a negative charge (eIII) reacted with polyglycerophosphate antisera from Streptococcus mutans and Streptococcus pyogenes. For comparison, the MT703 antigen in a hot trichloroacetic acid extract (eA) and the group E antigen from a saline extract of cells of strain K129 (EI) were similarly purified by anionic ion-exchange chromatography. Although the ratio of glucose and rhamnose in eA was 1:0.9 and in eI and eII approximately 1:1.5, reactions of identity were obtained in gel diffusion against specific anti-e serum. This difference in ratio is probably a result of the extraction procedures. Both the type e and group E antisera were reactive with both eI and EI antigens. The adsorption of group E antiserum with MT703 cells removed all E antibody, whereas type e-specific antibody remained after adsorption with K129 cells. These results suggest that eI antigen possesses both e and E specificities, whereas EI possesses E only. These findings were supported by the quantitative precipitin test and immunodiffusion and/or immunoelectrophoretic patterns in agar gel. Methyl-beta-D-glucopyranoside markedly inhibited the precipitin reaction in both type e and group E sera. However, a significantly stronger inhibition by cellobiose of type e serum than of group E serum indicates that a beta-linked glucose-glucose dimer is the predominant antigenic determinant of the e specificity. The presence of both e and E specificities on a single polysaccharide molecule was demonstrated by the use of purified e antigen released from a specific e-anti-e complex. This antigen reacted with a group E-specific serum as well as a type e-specific serum. An examination of five S. mutans type e strains showed the presence of group E specificity also, whereas the I, II, and IV serotypes of group E streptococci only possessed the group E specificity.


Immunity after infections with Myxoviruses. Influenza, parainfluenza and respiratory syncytial viruses cause respiratory infections in man with consequent transient and sometimes imperfect against reinfection. Humoral immunity and probably cell-mediated immunity contribute to resistance. Whereas circulating antibodies are more important for influenza viruses, secretory antibody are relatively speaking more important for parainfluenza and respiratory syncytial virsues. Measles and mumps induced longlasting immunity which can be correlated with circulating neutralizing antibodies. Certain immune responses against measles and respiratory syncytial virus cause pathological reactions after infection with the same virus.


Somatic cell hybrids between human lymphoma lines. II. Spontaneous and induced patterns of the Epstein-Barr virus (EBV) cycle. The regulation of spontaneous, IUDR-induced and P3HR-1 virus-induced EA and VCA production patterns was studied in two new somatic hybrids between human lymphoma lines. The hybrid 8A was derived from the crossing of the non-producer Raji with the spontaneous producer Daudi line. The second hybrid, 83, was produced by the fusion of Raji with the EBV-genome-negative B-lymphoma line, BJAB. The studies suggest the following EBV regulation patterns: (1) the spontaneous production of EA and VCA appears to be regulated by controls that differ from the regulators of P3HR-1 virus-induced or IUDR-induced EA synthesis. While spontaneous producer status was dominant over non-producer status, the level of EA inducibility was set by one of the parental cells, Raji ATG, and could either raise (in the previously studied Raji/Namalwa hybrid, cf Nyormoi et al. 1973) or depress (in Raji/Daudi) the level of relative EA inducibility found in the partner cell. (2) Although EA production is a prerequisite for VCA synthesis, the latter is under its own restriction mechanisms, quite independent of those that regulate the level of EA synthesis. (3) Inducibility of EA synthesis by P3HR-1 virus and by IUDR appear to be under the influence of at least partially identical controls. (4) EBV-negative lymphoma cells, exemplified by BJAB, may exert a "complementation" effect on the EA inducibility of their EBV-positive fusion partner, in spite of their own restrictivity against virus-induced EA synthesis. In more general terms, it is obvious that the EBV cycle is under the influence of multiple regulatory mechanisms in the human lymphoid cell. Depending on the parental cell and viral genomes that are allowed to interact, somatic cell hybrids may display a variety of patterns. At this time, cell hybridization is one of the few pathways that permit an approach to this complex and completely unknown world.


Cell line-specific cell surface components of hypotetraploid ascites mammary carcinoma cells inducing humoral reactions in the syngeneic host. Cell-line specific cell surface components were demonstrated on cells of various hypotetraploid ascites cell lines, but not on hyperdiploid cells derived from a mammary carcinoma induced by mammary tumor virus (MTV) in syngeneic C3H/He mice. These substances caused cell-line-specific transplantation antigenicity and were identified, together with MTV-associated substances and tumor-associatied embryonic materials, as being the binding sites for tumor cell agglutinating factors found in sera from tumor-bearing and regressor animals. Particles containing the cell line-specific substances were released from these cells by hypnotic treatment. They could be purified as a single peak in a sucrose density gradient. They were, however, not dissociated from the MTV-associated substances. Absorption studies of agglutination activity were extensively used for demonstration of specificity.


Specific antigen of murine erythroblasts. A new antigan has been revealed by means of antisera against Rauscher virus in mice with Rauscher virus-induced leukemia. This antigen appears to be different from both Rauscher type-specific antigen and MULV-gs-1 (p-30), as shown by studies of electrophoretic mobility and immunochemical specificity. Except in leukemic mice it was also found in low levels in both serum and spleen extracts of healthy mice of a number of strains. Furthermore, this antigen was regularly demonstrated by immunofluorescence on the surface of erythroblasts, but not on the surface of erythrocytes, lymphocytes, polymorphonuclear cells and thymocytes, and was shown to be different from fetal hemoglobin. Therefore, it is referred to as antigen of erythroblasts (Ag-Eb), which seems to represent a surface marker for a certain differentiation stage of erythroid cells.


Tube leukocyte (monocyte) adherence inhibition assay for the detection of anti-tumour immunity. III. "Blockade" of monocyte reactivity by excess free antigen and immune complexes in advanced cancer patients. Leukocytes from patients with limited cancer display LAI reactivity whereas leukocytes from patients with metastatic cancer frequently demonstrate no reactivity in the tube LAI assay. The leukocytes (monocytes) of reactive patients react with tumour antigen through specific cytophilic anti-tumour IgG antibody bound to the monocyte's Fc cell surface receptors. The non-reactive monocytes from patients with advanced cancer lacked the ability to bind free cytophilic anti-tumour antibody. Moreover, the serum of the non-reactive patient contained no free cytophilic anti-tumour antibody capable of "arming" normal leukocytes. The serum of patients with large tumour burdens contained free tumour antigenic determinants capable of absorbing free cytophilic anti-tumour antibody from the serum of reactive patients or when preincubated with reactive leukocytes abrogating their LAI responsiveness immunologically specifically. Blocking was immunologically specific; therefore, the specificity must reside in the tumour antigenic determinant since immune complexes are bound nonspecifically. The tumour antigen coat was removed by gentle trypsinization of the monocyte's surface. This restored the monocyte's capacity to react with the sensitizing tumour antigen and to bind free cytophilic antibody from the microenvironment. Nonreactivity in the tube LAI assay of patients with metastatic cancer was not the result of a numerical deficit of circulating monocytes but was mediated by an excess of tumour antigen in the microenvironment of the sensitized monocyte.


Identification with G and R banding of the position of breakage points induced in human chromosomes by in vitro x-irradiation. The aberrations seen in chromosomes of human peripheral-blood lymphocytes, X-irradiated in vitro, have been analysed in three types of preparations, treated to give G-banding; R-banding; and G-banding followed by R-banding on the same cells. The data from cells subjected to both banding techniques reveals that 30 per cent of the sites of chromosome breakage are situated at he interfaces between dark- and - light-stained bands. The results of all the analyses show that approximately 30 per cent of all breaks were located in either the telomere (19-5 per cent) or centromere (11-3 per cent) regions. Chromosomes rich in R-band material were not preferentially damaged, but chromosomes 12, 15, and particularly 17, were involved in aberrations more frequently than would be expected on the basis of their length. No breaks were found on the Y chromosome in the 114 male cells analysed, but the X did not appear to be spared from damage either in the male cells analysed, but the X did not appear to be spared from damage either in the male cell or the 136 female cells analysed. G and/or R-banding enables a much more accurate analysis of aberrations than can be obtained from the use of conventional staining techniques, and with these methods, it is shown that the numbers of induced asymmetrical and symmetrical exchanges are similar.


Immunity against avian sarcomas: a review. Animals bearing either primary or transplantable tumors of avian sarcoma virus origin frequently mount cellular and humoral antitumor immune responses against both viral and nonviral antigenic determinants. This communication summarizes recent developments in this area, with special emphasis on the effector mechanisms of antitumor immunity in chickens. Among the topics considered are structural and antigenic aspects of avian leukosis and sarcoma viruses; virus-induced tumor-associated antigens; methods of in vivo and in vitro detection of tumor-associated antigens; and the relative importance of the bursal and thymic systems in avian antitumor immunity.


Lysis and necrosis: analysis of two cytotoxic phenomena mediated by lymphocytes. Stimulation of rat lymphocytes by allogeneic and xenogeneic (mouse) embryo fibroblast monolayers or by pokeweed mitogen (PWM) generate two different cytotoxic manifestations: a) cell-mediated target cell lysis obtained by contact between effector cells and target cells; b) cytotoxicity caused by lymphotoxin (here termed "necrosis"). Activation of hypersensitive lymphocytes by soluble antigen, e.g., keyhole limpet hemocyanin (KLH), produces only necrosis but no lytic effect. This difference between mitogen and antigen activation is attributed to the affinity of the mitogens for target cell membranes. There is a marked difference in the time of onset of the two phenomena. Ten minutes after the effector cells have been introduced to the target monolayers, lysis of fibroblasts becomes visible, whereas necrosis usually develops only 48 to 72 h later. A clear difference in the morphology of the two types of target cell killing was seen when the cultures were stained in situ with trypan blue. Lysed target cells produced blebs and underwent fragmentation, scattering cytoplasmic droplets. During a 10-min to 24-h period, these cells excluded the dye. Only much later (after greater than 30 h), when the cell remnants had undergone a further slow "decay" process, did they stain with trypan blue. In cultures rendered necrotic by lymphotoxin, the killed fibroblasts preserved their morphological integrity but the nuclei became markedly stained with trypan blue. Lymphotoxin was found to be effective only in highly concentrated macrophage and fibroblast cultures. In sparsely populated macrophage cultures, the mediator(s) did not effect cell death but nonetheless exerted marked morphological changes. Vacuoles containing lipids developed and eventually occupied the greater volume of the cytoplasm. Giant lipid droplets were seen leaking out of the macrophage cytoplasm. It is suggested that lymphotoxin does not kill directly, but rather that metabolic products released by the vigorously stimulated target cells cause the necrotic effect in the cultures.


Localization of glycoidases with naphthyl substrates. Unsubstituted naphthyl substrates were found to be superior to substituted naphthyl, indolyl and hydroxyquinoline substrates for the histochemical demonstration of alpha-mannosidase, alpha-galactosidase, hetero-beta-glycosidase, glucoamylase and sucraseisomaltase, equivalent for beta-N-acetylglucosaminidase and lactase-beta-glucosidase, and inferior for beta-glucuronidase and acid beta-galatosidase. Aldehyde fixation is necessary for the localization of lysosomal glycosidases with naphthyl substrates. 1-naphthyl substrates are suitable for the detection of acid glycosidases in lysosomes and hetero-beta-glysocidase in the cytoplasm of animal cells, and 2-naphthyl substrates can be employed for the demonstration of microvillous glycosidases and for the evaluation of the total activity of soluble glycosidases with semipermeable membranes. When naphthyl substrates are used coupling should be carried out simultaneously and hexazotized pararosaniline is the coupling reagent of choice.


A novel approach for enzyme histochemical and autoradiographic studies on single cells. A simple technique that does not involve the use of heat has been developed to fix cells or cell organelles. The cells or organelles are mixed with a bovine serum albumin solution, gelled by the addition of a suitable fixative, and then either embedded or frozen. The gelled mixture contains well preserved cells or organelles that are evenly dispersed, thus eliminating the problems of pellet packing. The technique was excellent for ultrastructural autoradiography where radioactive materials bound to plasma membranes or cytoplasmic nucleotides were being studied. Histochemical tests could be applied to the fixed embedded material. Light and electron microscopy could be done on the same well-mixed sample. Fixed frozen albumin samples cut with ease on a cryostat but there was ice crystal formation.


Further characterization of phosphatase activities using non-specific substrates. The demonstration of non-lysosomal acid phosphatase has been the subject of a number of recent investigations. In the present study we compared the enzyme activities in rat liver and kidney that are revealed after incubation in the presence of either beta-glycerophosphate, p-nitrophenylphosphate or phenylphosphate at varying pH. As seen by others, the activity towards p-nitrophenylphosphate at pH 5-6 was confined to lysosomes, Golgi apparatus, endoplasmic reticulum (ER), nuclear envelope and plasmalemma. The reactivity of the plasmalemma and the ER was increased at pH 7. The TER of Küpffer cells in the liver stained intensely in contrast to the ER of the parenchymal cells, which stained only weakly. In the presence of NaF, all sites except the plasmalemma became negative. Addition of a levamisole-analogue, L-p-bromotetramisole, which is a specific inhibitor of alkaline phosphatase, resulted in the disappearance of the plasmalemmal activity whereas the activity at the other sites appeared unaltered. The rather unusual locations of activities with so-called non-specific substrates were further compared with those obtained with specific substrates such as glucose-6-phosphate and thiamine pyrophospate. The possible implication of these data in relation to the specificity of marker-enzymes for subcellular organelles is discussed.


Use of the p-nitrophenyl phosphate method for the demonstration of acid phosphatase during starvation and cell autolysis in the planarian Polycelis tenuis Iijima. Acid phosphatase activity is demonstrated employing p-nitrophenyl phosphate as substrate and lead acetate as coupler. The fine structural localization of the enzyme in starved planarian tissues is described. The method is used to pin-point starvation - induced acid phosphatase activity in relation to autophagy and crinophagy in the gland cells; autophagy, autolysis and cell death in parenchymal and gastrodermal cells and basement membrane lysis. Attention is also payed to the demonstration of muscle lysis. The histochemical implications of the method are discussed.


[Ultrastructural identification and immunocytochemical study of somatostatin cells in antral mucosa of the rabbit and the mouse (author's transl)]. In normal and L-Dopa treated rabbits and mice, combined immunochemical methods, photonic histological methods for endocrine cells and ultrastructural methods were used to elucidate ultrastructure and properties of somatostatin cells of the antral mucosa. In normal rabbits, immunoreactive cells giving no fluorescence with Falck's technic, they corresponded neither to serotonin cells nor gastrin cells; they were unreactive with Fontana, Hellerström-Hellmann, Sevier-Munger and Mac Conaill methods but very slightly stained with Grimelius methods. In L-Dopa treated animals somatostatin cells gave formaldehyde induced fluorescence (they were included in GIC cells, thus in Apud group), exhibited a good reaction with Grimelius and Sevier-Munger methods. In order to carry out the alternate semi-thin/thin section procedure (semi-thin sections for immunofluorescence or immunoenzymatic detection and serial thin sections counter-stained for conventional ultrastructure studies), immunological treatment were performed on M.F.F.--glutaraldehyde fixed small fragments of mucosa before inclusion in Epon 812 or, after inclusion, on semi-thin sections. We succeeded in identifying ultrastructurally somatostatin cells. They displayed round or ovoïd shaped secretory granules, and three constant typical structures: numerous microfilaments--light and homogenous granules, often seeming like lipids---granules made up by coarsely filamentous cores surrounded by a large empty halo. Somatostatin cells seemed different of X cells because of their predominant localisation in the antral mucosa (in the rabbit X cells were predominantly in the fundus) and because of the lack of nuclear microfilaments; they also seemed ultrastructuraly different of D1 cells.


Alkaline lead citrate: a selective stain for glutaraldehyde precipitates of indolamines and primary catecholamines in electron microscopy. Test-tube experiments proved that alkaline lead citrate (Reynolds, 1963), which is generally used as an electron-opaque stain, specifically reacts with precipitates formed by glutaraldehyde and biogenic amines (indolamines, primary catecholamines). Glutaraldehyde/osmium tetroxide fixation and staining of thin sections with alkaline lead citrate is recommended as an optimal preparation method of studying the above-mentioned amines at a fine structural level.


Rapid nuclear staining method for Saccharomyces cerevisiae. Mithramycin was used to stain nuclei in mitotically dividing and sporulating yeast.


Platelet alpha2-macroglobulin and alpha1-antitrypsin. Subcellular membrane and granule fractions derived from human platelets contain immunologically identifiable alpha2-macroglobulin and alpha1-antitrypsin. These platelet-derived inhibitors show a reaction of immunologic identity when compared to alpha2-macroglobulin and alpha1-antitrypsin purified from human plasma. Further, the platelet protease inhibitors possessed a similar subunit polypeptide chain structure to their plasma counterparts as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis. Studies of the binding of radiolabeled trypsin to the various solubilized platelet subcellular fractions suggest that the granule-associated alpha2-macroglobulin and alpha1-antitrypsin, as well as membrane-associated alpha2-macroglobulin were functionally active. Quantitatively, circulating platelets contain relatively small concentrations of these inhibitors as compared to platelet-associated fibrinogen and factor VIIIAGN. Platelet protease inhibitors may modulate the protease-mediated events involved in the formation of hemostatic plugs and thrombi.


A fragment comprising the last third of bovine serum albumin which accounts for almost all the antigenic reactivity of the native protein. The fragmentation of native bovine serum albumin by trypsin has been studied in aqueous solution under various conditions with regard  to the yield and size of the fragments obtained. From a partial tryptic hydrolysate at pH 8.2 (40 degrees, 1 hour), a homogeneous fragment was isolated in high yield by gel filtration on Sephadex G-100, followed by chromatography on DEAE-cellulose. The molecular weight of the fragment by gel filtration on calibrated Sephadex G-100 columns and by sodium dodecyl sulfate electrophoresis was 22,500. After reduction of the disulfide bonds followed by alkylation of the resultant thiol groups with iodoacetamide, the fragment retained homogeneity by disc electrophoresis and its molecular weight remained unchanged, indicating that it was composed of a single polypeptide chain. From its amino acid composition, sequence of the first 20 residues, and actions of carboxypeptidases A or B, it was unequivocally assigned to positions 377-571 in albumin. The inhibitory activity of the fragment was 90 to 93% towards the immune reaction of the protein with the IgG fraction of the antisera. The IgGfraction accounted for 96% of the total antibody activity in the antisera. An immunoabsorbent of fragment 377-571 removed 89 to 95% of the antibody to albumin. A fluorescent derivative of the fragment, which retained full immunochemical activity, was found to bind 2 mol of antibody/mol of peptide. The disulfides in peptide 377-571 were essential for its immunochemical reaction because the latter was entirely abolished upon reduction and S-alkylation of the disulfides. Since this fragment comprised only a third of the albumin molecule, but accounted for 90 to 95% of its antigenic reactivity, the results indicated that native albumin carries identical repeating antigenic reactive sites.


The beads in the Golgi complex-endoplasmic reticulum region. The region between the rough endoplasmic reticulum (ER) and the Golgi complex has been studied in a variety of insect cell types in an attempt to find a marker for the exit gate or gates from the ER. We have found that the smooth surface of the rough endoplasmic reticulum near Golgi complex transitional elements has beadlike structures arranged in rings at the base of transition vesicles. They occur in all insect cell types and a variety of other organisms. The beads can be seen only after staining in bismuth salts. They are 10-12 nm in diameter and are separated from the membrane and one another by a clear halo giving them a center to center spacing of about 27 nm. The beads are not sensitive to nucleases under conditions which disrupt ribosomes or remove all Feulgen staining material from the nucleus. Under conditions similar to those used to stain tissue, bismuth does not react in vitro with nucleic acids. The component of the beads that stains preferentially with bismuth is therefore probably not nucleic acid.


Thyrotoxicosis secondary to involvement of the thyroid with malignant lymphoma. A patient with malignant lymphoma developed goiter and thyrotoxicosis during the course of her disease. A thyroid biopsy revealed involvement of the thyroid gland with a malignant lymphoma. This was associated with the high levels of circulating thyroglobulin and thyroid hormones. The patient was treated with propylthiouracil, local radiotherapy, and nitrogen mustard and prednisone. The patient became euthyroid with the disappearance of goiter. Circulating levels of thyroglobulin and thyroid hormones returned to the normal range.


Influence of purified plasma proteins on testosterone uptake and metabolism by normal and hyperplastic human prostate in "constant-flow organ culture". Surgical samples of human prostate were explanted and submitted to constant-flow organ culture. The medium contained 3H-testosterone 50 nM, and except for controls, increasing concentrations of human serum albumin (HSA) or human sex-steroid-binding plasma protein (SBP). At steady state, the explants were washed and homogenized, and the total radioactivity, radioactive testosterone, androstanolone (17 beta-hydroxyandrostan-3-one), androstane-3 alpha, 17 beta-diol, and androstane-3 beta, 17 beta-diol were determined after the addition of the corresponding internal 14C standards. From these data, testosterone uptake and metabolism were quantitated. The concentration of unbound testosterone in protein-supplemented culture media was measured separately by equilibrium dialysis. In control superfusions without protein, the tissue concentration of total radioactive steroids was equivalent to 182 +/- 18 (mean +/- SEM) pmoles/g of prostate. Androstanolone represented about 2/3, testosterone 1/10, and the two androstanediols together 1/10 of the total radioactivity. No difference was found between "normal" and hyperplastic prostate explants. In experiments with HSA (15-176 muM), is was observed that the uptake of radioactive testosterone in the prostate explants was decreased in direct proportion to the unbound testosterone fraction of the superfusion medium, but the proportions of testosterone metabolities in the superfused explants remained the same. In experiments with SBP (6-135 nM), the concentrations of unbound testosterone in the superfusion medium were reduced to the same levels as in the experiments with HSA. The reduction of tissue radioactivity was somewhat larger than that expected from the reduction of unbound testosterone in the superfusion medium for the concentrations of SBP less than 50 nM, and then remained approximately constant. In addition, SBP altered the metabolism of testosterone: the androstanolone/testosterone ratio in the prostate explants was critically dependent upon the SBP concentration in the superfusion medium. It is therefore suggested that, independent of its effect on the binding of testosterone, SBP has a direct effect on testosterone uptake and metabolism by the human prostate. The underlying mechanism is unknown.


Pituitary-thyroid regulation in euthyroid patients with Graves' disease previously treated with antithyroid drugs. To understand why some patients with hyperthyroidism due to Graves' disease remain euthyroid after a course of antithyroid drug therapy, pituitary-thyroid regulation was studied in 20 such patients who had remained well for six months or longer after the withdrawal of antithyroid drugs. Only patients who were clinically euthyroid and had normal serum thyroxine (T4), triiodothyronine (T3), and thyrotropin (TSH) concentrations were studied. Serum TSH responses to thyrotropin-releasing hormone (TRH) and thyroid suppression were determined in all patients. Seven patients had normal responses to both tests. Six patients had a subnormal response to TRH and abnormal suppression. Five patients had a subnormal response to TRH and normal suppression, and two patients had a normal TSH response to TRH and abnormal suppression. There were no differences in the mean serum T4, T3 or TSH concentrations between any of the groups. The mean duration of time after antithyroid drug withdrawal was 19 months in the patients in whom both tests were abnormal, whereas it was 58 months in those in whom both tests were normal and 45 months in those with a subnormal TSH response to TRH and a normal suppression test. Thus, in 13 of the 20 patients studied, various degrees of abnormality of pituitary-thyroid regulation were demonstrable. These results suggest that, in most patients with Graves' disease who remain clinically and biochemically euthyroid after a course of antithyroid drug therapy, the disease persists in a mild or subclinical form.


Delayed hypersensitivity cross-reactions between Sporothrix schenckii and Ceratocystis species in sporotrichotic patients. Cutaneous delayed hypersensitivity to antigens prepared from Sporothrix schenckii and several Ceratocystis species, including C. stenoceras, C. ulmi, C. ips, and C. minor, was tested in 14 patients with known cutaneous sporotrichosis. Extensive cross-reactions were observed. Nonsporotrichotic people (controls) did not react to these antigens. The correlation coefficient between antigens of S. schenckii and each Ceratocystis species was calculated from the areas of the cutaneous reactions. Among the Ceratocystis species tested, the correlation coefficient between S. schenckii and C. stenoceras was 0.91.


Serological studies and isolations of serotype hardjo and Leptospira biflexa strains from horses of Argentina. Three pathogenic leptosipras and 12 saprophytic Leptospira biflexa strains were isolated from 72 apparently normal horse kidneys collected at an abattoir in Argentina. Cross-agglutination reaction patterns of the pathogens showed that they were antigenically homologous with members of the Hebdomadis group. When one of the strains was compared to Hebdomadis serotypes in reciprocal agglutination-absorption tests, it was found to be serologically homologous to serotype hardjo. This is the first known report of an isolation of this serotype from horses. Serological tests were also carried out on randomly collected abattoir sera from 245 horses to determine the prevalence of equine leptospirosis. Significant antibody titers (1:100 or greater) were found in 74.6% of the sera. Predominant reactions occurred with the antigens pomona, hebdomadis group, pyrogenes, tarassovi, and canicola. Agglutination tests performed with antigen prepared with one of the saprophytic biflexa isolates showed seropositive reactions in 99.1% of the equine sera, with agglutination titers ranging from 1:100 to 1:3,200. Absorption of selected horse sera with the saprophytic strain removed the agglutinins to Leptospira interrogans serotypes. This suggests the possibility that L. biflexa strains may act as an antigenic stimulus and account for some of the persistent multiple cross-reaction patterns of equine sera with pathogenic serotypes.


A simplified Leifson flagella stain.

A simplified Leifson flagella stain. Flagella of bacteria taken directly from 24- and 48-h blood agar plates can be stained by using a simple, reliable procedure.


IgA and IgG anti-ragweed antibodies in nasal secretions. Quantitative measurements of antibodies and correlation with inhibition of histamine release. Total secretory IgA and specific anti-antigen E (AgE) antibodies (ab) in the IgA and IgG classes were measured in concentrated nasal washings from ragweed allergic and normal individuals by antigen binding or anti-alpha-radioimmunoassays. Virtually all the allergic patients had significant IgA (45/49) and IgG (46/49) ab to AgE in their nasal washings. By contrast, washings from most normal persons contained no measurable IgA (13/15) ab or IgG (13/15) ab to AgE. The total IgA levels in allergic washings were not significantly different from those in normal washings and they were used to standardize the ab measurements. Parenteral immunotherapy with ragweed extract increased specific nasal IgA ab from 10.6 +/- 2.7 (SEM) to 39.0 +/- 8.7 ng AgE bound/mg IgA and IgG ab from 17.2 +/- 2.6 to 65.1 +/- 7.4 ng AgE bound/mg IgA (P less than 0.001 for both classes). The ratio of IgA:IgG ab was not affected by therapy, and for treated patients, there was no correlation (rs + 0.32, P greater than 0.1) between nasal IgG ab and serum IgG ab. These results suggest that at least part of the nasal IgG ab is produced locally. Blocking activity in the nasal washings was measured by inhibition of histamine release and was found to correlate directly (rs + 0.85, P less than 0.001) with binding activity for AgE. Some washings from normal persons caused slight inhibition of histamine release but others caused enhancement. Nasal washings were fractionated by passage over Sephadex G-200. Inhibition of histamine release by dilutions of the IgA-rich and IgG-rich fractions correlated well with binding activity in these fractions. None of these results support the hypothesis that allergic individuals are deficient in secretory IgA or secretory ab responses. These results, however, are in keeping with the theory that hay fever occurs in a high-responder population which is genetically able to respond to low doses of inhalant antigens.


Modulation of cellular-immune responses in vivo and in vitro by histamine receptor-bearing lymphocytes. Histamine, one of the mediators involved in the IgE-mediated reaction, was demonstrated to influence in vivo and in vitro components of cellular-immune reactions in orthochlorbenzoyl-bovine gamma globulin-immune guinea pigs. 10(-3) M histamine reduced by half the size of a delayed hypersensitivity skin test at 24 h. Inhibition of skin reactivity by histamine could be partially reversed by H-1 receptor antagonists such as chlorpheniramine and completely prevented by H-2 receptor antagonists such as burimamide. The histamine suppression of cutaneous delayed hypersensitivity could be accounted for in part by its inhibitory effect on certain lymphocyte responses including antigen-induced migration inhibitory factor (MIF) production and proliferation. At concentrations of 10(-3)-10(-5) M histamine reversibly inhibited MIF production and its action could be blocked by H-2 antagonists but not H-1 antagonists. Thus, lymphocytes bearing H-2 receptors modulate MIF production and probably lymphocyte proliferation as well. Histamine did not interfere with the macrophage response to preformed MIF. These studies indicate that immediate hypersensitivity reactions involving histamine release might influence the subsequent expression of cellular-immune reactions.


Rosette formation between human lymphocytes and sheep erythrocytes. Inhibition of rosette formation by specific glycopeptides. Rosette formation with unsensitized sheep erythrocytes is a characteristic of human thymus dependent lymphocytes. Release of glycopeptides from the sheep erythrocyte by trypsin reduces rosette formation. These tryptic glycopeptides inhibit rosette formation by untrypsinized sheep erythrocytes; this suggests that rosetting is mediated by erythrocyte surface glycopeptides. To investigate the molecular nature of this interaction, we examined the abilities of various model compounds to act as haptenic inhibitors of rosette formation. Inhibition is given by glycopeptides bearing oligosaccharide units rich in sialic acid, galactose, N-acetylglucosamine, and mannose linked to asparagine residues through glycosylamine bonds. Among compounds tested, fetuin glycopeptide is most effective, but human transferrin glycopeptide and human erythrocyte glycopeptide I also inhibit rosette formation. Other compounds including human erythrocyte glycopeptide II, human IgG glycopeptide, lacto-N-neotetraose, 3'- and 6'-sialyllactose show no significant inhibition. Neither sialic acid, galactose, manose, nor N-acetyl-glucosamine alone inhibits rosette formation. Stepwise degradation of fetuin glycopeptide established the galactose residues as important determinants of inhibitory activity. Fetuin glycopeptide blocks rosette formation when added to a suspension of human lymphocytes and sheep erythrocytes or when preincubated with human lymphocytes, but not when preincubated with sheep erythrocytes. Studies of the binding of [3H] fetuin glycopeptide to normal lymphocytes demonstrate 7.5 x 10(6) saturable binding sites per cell. No saturable binding of this compound to sheep erythrocyte membranes is observed. Compared to normals, lymphocytes from patients with chronic lymphatic leukemia demonstrate decreased fetuin glycopeptide binding with a mean of 0.9 x 10(6) sites per cell. This decreased binding correlates with the impaired ability of these cells to form rosettes. The data suggest that fetuin glycopeptide inhibits rosette formation by binding to the thymus-dependent cell where competition occurs with sheep erythrocytes for specific lymphocyte surface receptors.


The induction of prostatic hypertrophy in the dog with androstanediol. The effects of androstanediol and estradiol on prostatic growth were investigated in castrate dogs. Estrogens along resulted in no significant change in prostatic weight, whereas androstanediol produced growth comparable to that in uncastrated controls. Androstanediol plus estradiol resulted in an even more striking increase in prostate growth. Approximately half the animals receiving androstanediol alone and all of those receiving androstanediol plus estradiol fulfill the weight and histological criteria for prostatic hypertrophy in the dog. Since both these steroid hormones are presumed to be normal secretory products of the testis, it is possible that they are involved in the pathogenesis of prostatic hypertrophy in the dog.


Evaluation of the time spent in learning from a programed text, a slide-tape, and a lecture at six dental schools. 1. Programed text and slide-tape presentations required significantly less study time than the lecture method. 2. Students of varying academic ability required approximately the same amount of study time. 3. Familiarity with self-paced study methods did not significantly affect study time.


In vivo and in vitro correlates of food allergy. Sera of 86 patients clinically sensitive to foods were tested by passive sensitization of human and/or monkey lung (127 tests) and the radioallergosorbent test (RAST) (72 tests), using whole-food antigens; the results were compared with skin (prick) testing. Results of the prick test correlated with history in 76% of cases; lung sensitization correlated with history in 37% and with prick test in 57%; and RAST correlated with history in 54% and prick test in 72%. It is concluded that a very large percentage of adverse reactions to foods are IgE-mediated. The prick test is of use in diagnosis, particularly when combined with RAST; the lung sensitization test is technically impractical and not a reliable indicator. The best diagnostic method is careful history with food challenge and withdrawal and rechallenge; the latter is safe except in patients with a history of violent reaction.


Studies of the antigenicity and allergenicity of phospholipase A2 of bee venom. The antigenic and allergenic properties of phospholipase A2 (PLA2) and whole bee venom were compared by measuring the IgG and IgE antibody responses in animals and man. Precipitating antibodies raised in rabbits and reaginic and other antibodies raised in mice reacted about equally with both bee venom and PLA. The majority of human sera containing bee venom-specific IgE also contained PLA-specific IgE, although in somewhat lower titers. Similarly, most human sera with significant amounts of total antibodies reacting with bee venom also had antibodies reacting with PLA. Histamine and SRS-a release from leukocytes of sensitive patients followed challenge with whole bee venom and PLA in the majority of instances. However, mediator release from several patients' cells was obtained with bee venom only. These studies suggest that although PLA is a major allergen and antigen in bee venom, significant exceptions in patients' reactivity may limit its potential diagnostic and therapeutic usefulness.


The effects of input condition on free recall in young and old adults. Free recall lists were presented to young (17 to 19, N = 42) and old (60 to 70, N = 42) males under three conditions: (a) active auditory - subjects read words aloud; (b) passive auditory - experimenter read words aloud as shown; (c) visual only - no one read words aloud. For words from the end of a list, recall was best at the active-auditory condition and worst at the visual-only condition for both age groups. For words from the beginning of a list, recall was best at the visual-only condition and worst at the active-auditory condition for both age groups. The young group recalled more words than the old at each presentation position under each condition.


Histochemical characteristics of the striated inclusions of adrenoleukodystrophy. The straited accumulations in adrenal cortical cells and brain macrophages that are characteristic of adrenoleukodystrophy have been studied histochemically in cryostat sections to seek leads for the biochemical identification of the striated material. It stained pale pink with oil red O and did not stain with the Schultz cholesterol procedure or periodic acid-Schiff technique. By utilizing the birefringence of the accumulations as a marker, it was determined that, unlike natural cholesterol and cholesterol esters, the striated material was resistant to acetone and ethanol extraction. It was readily soluble, however, in nonpolar solvents such as n-hexane and chloroform. These findings indicated that the material was most probably a lipid, and they suggested that sequential extraction of adrenoleukodystrophy adrenal and brain with acetone and then n-hexane could be used to isolate this material in relatively pure form. Based on this lead, biochemical studies have just revealed a fatty acid abnormality in adrenoleukodystrophy which appears to be unique to this genetic disease.


X-ray microanalysis of zinc and calcium in ultrathin sections of human sperm cells using the pyroantimonate technique. X-ray microanalysis of pyroantimonate-fixed sperm cells indicates the retention of calcium and zinc subcellularly in similar proportions to air dried cells. The ultrastructure is well preserved and is corelated with the analysis. Sodium, potassium and chlorine are all removed during the fixation. CAlcium and zinc are found present intracellularly both in association with and independent of antimonate precipitation. There thus appears to be a varying degree of binding of those elements subcellularly, precipitation occurring where binding is reduced.


Origin and significance of centric fusions in domestic sheep. The karyotypes of 731 sheep of various breeds were studied and considered in association with previous chromosome studies of domestic sheep. A high incidence of the t2 trnslocation was found in two pedigree flocks of New Zealand Romney sheep. One of these flocks was established over 100 years ago and it is suggested that this translocation originated in the Romney Marsh breed of sheep in England. A naturally occurring double translocation heterozygote 52t1t2 was reported for the first time. A further flock of sheep of the Perendale breed was found with a high incidence of dicentric chromosome fusion that was identified as the t3 translocation. The apparently common occurrence of chromosome polymorphism, due to centric fusions, in domestic sheep is discussed in relation to karyotype evolution among both domestic and wild sheep.


Isolation and preliminary characterization of two varieties of low molecular weight immunoglobulin in the bullfrog, Rana catesbeiana. Two varieties of low m.w. immunoglobulins have been isolated from the serum of Rana catesbeiana frogs. They are highly cross-reactive, although each also contains unique antigenic determinants. Since both low m.w. immunoglobulins were identified in the serum of 22 individual frogs, it was concluded that they are isotypic variants. The light chains of R. catesbeiana and mammalian high and low m.w. immunoglobulins are similar in electrophoretic mobility on polyacrylamide gels containing sodium dodecyl sulfate. The heavy chains of fropg high m.w. immunoglobulins have the mobility of mammalian mu-chains; the heavy chains of both variants of frog low m.w. immunoglobulins migrate between mammalian mu- and gamma-chains in approximately the position of mammalian alpha-chains. An unusual structural feature of the R. catesbeiana high ald low m.w. immunoglobulins is that the unreduced proteins are partially dissociated in sodium dodecyl sulfate.


Staining technique for definite identification of individual cattle chromosomes in routine analysis. The purpose of the present communication is to draw attention to the great applicability of a recently introduced R-banding technique for definite identification of cattle chromosomes in routine analysis. Banding patterns as well as densitometric measurements of normal bull chromosomes and the so-called 1/29 translocation are demonstrated.


Cross reactivity of human beta2-microglobulin with human granulocyte colony-stimulating activity. Effects of antisera to human beta2-microglobulin (beta2 m) on factors able to stimulate colony formation in culture by human granulopoietic progenitor cells were investigated. The colony-stimulating activity (CSA) present in media conditioned by cultures of human peripheral leukocytes was suppressed by treatment with anti-beta2m. This inhibition was not due to a direct effect on the granulopietic progenitor cells; controls to test for cytotoxicity and for noncytotoxic inhibition of the progenitor cells by anti-beta2m yielded negative results. These experiments provide evidence for a relationship between human CSA and beta-microglobulin, and suggest a possible analogy between molceules involved in the in vitro regulation of granulopoiesis and products of the major histocompatibility gene complex.


Cross-reactive T-cell antigens among mammalian species. Fluoresceinated heteroantisera prepared against T cells of rats, monkeys, and humans were reacted withthymus and spleen cells from 11 selected species. These reagents recognized cross-reacting T cell antigen(s) among rodent species (mouse, rat, guinea pig, and hamster) and among primate species (monkey and humans). With one exception, the cross-reactivity was restricted to a phylogenetic order. All three antisera required relatively few absorptions to achieve T cell specificity for related species when compared to absorption requirements for the isologous species. Differentiation antigens within a phylogenetic order thus appear to be more homologous than other cell surface constituents on T cells.


Complement-mediated release of histamine from human basophils. II. Biochemical characterization of the reaction. Release of histamine from human basophils was induced by activation of complement using zymosan. The histamine-releasing factor resembled C5a on the basis of m.w. (15,000) as well as previous studies showing inactivation by anti-C5. Complement-induced release of histamine was compared with allergic release of histamine which is mediated through appropriate allergens and reaginic IgE. Previously we demonstrated that the former reaction occurred more quickly. Both reactions were inhibited by drugs which increase intracellular concentrations of cAMP3 (theophylline, prostaglandin E1, and histamine) or which mimic the action of cAMP (its dibutyrly derivative). Calcium was required for complement-mediated release of histamine and an increasing response was observed up to physiologic concentrations (2 mM). Magnesium (0 to 1 mM) did not affect the amount of histamine released. Also, glycolysis was probably required for optimal release by complement, since both 2-deoxyglucose and iodoacetamide were inhibitory. When basophils were partly enriched by depletion of neutrophils and eosinophils, the percentage of histamine released by complement was unchanged. Finally, it was shown that activated complement desensitized basophils from responding to a second challenge by the same stimulus. Cross-desensitization was not observed between complement and pollen allergens.


Studies on the genetic control of antibody affinity: the independent control of antibody levels and affinity in Biozzi mice. The levels (Abt) and relative affinity (KR) of antibody produced to protein antigens injected in saline have been measured in the 22nd generation of the Biozzi high (Ab/H) and low (Ab/L) antibody-producing mice. No significant differences in affinity were observed between the two lines of mice (p 0.10) but the levels of antibody (Abt) differed significantly (p 0.0025) when immunized with antigen in saline; however, both Ab/H and Ab/L mice were able to mount a high affinity response to protein antigens injected in Freund's complete adjuvant. These results substantiate earlier observations that in mice, antibody affinity (KR) and antibody level (Abt) are under independent genetic control.


Cold activation of complement i. presence of coagulation-related activator. Determination of the complement titer in the serum and plasm of 120 patients with chronic liver diseases showed that in eight (7%) patients with cirrhosis of the liver, chronic active or chronic inactive hepatitis complement in the serum was less than half in the plasma. The dissociation of complement serum and plasma was due to cold activation of the classical pathway of complement in vitro since serum drawn from these patients at 37 degrees C lost hemolytic activity in 4 hours when transferred to a cold environment. Neither HB antigen nor cryoglobulin participated in this phenomenon. The activation of complement in the cold could be prevented by increasing the ionic strength, or by adding vitamin E or, to a lesser extent its vehicle HCO-60, while heparin, Trasylol, soybean trypsin inhibitor, or hirudin had no effect. Trans-AMCHA prevented activation in one case. It is speculated that a factor appearing as a result of blood clotting is able to activate the classical pathway of complement in the cold; it is probably not related to Hageman factor (factor XII), factor VII, thrombin, kallikrein.


Analysis of the diversity of murine antibodies to dextran B1355. I. Generation of a larger, pauci-clonal response by a bacterial vaccine. Mice immunized with a combination of dextran B1355 in adjuvant followed by three injections of 2 x 10(9) Escherichia coli B organisms produced an average of 14.5 mg/ml of anti-dextran antibodies. It was demonstrated that the stimulating effect of E. coli B was due to antigenic determinants cross-reactive with B1355 and not solely because of adjuvant properties of the organism. The anti-dextran antibodies were distributed among both 7S and 19S components. Isoelectric focusing of the 7S antibodies showed several spectrotypes of antibody, most of which were shared by the majority of the individual sera. The limited spectrotypic heterogeneity of the 7S antibodies was supported by idiotypic studies. Thus, a heterologous, anti-idiotypic serum, rabbit anti-M104, was prepared which distinguished between two closely related myeloma proteins, M104 and J558,with specificity for alpha-(1 leads to 3) dextran. This antiserum demonstrated that some, but not all, of the 7S and 19S anti-dextran antibodies possessed variable region determinants cross-reactive with M104.


Histamine release by chemotactic, formyl methionine-containing peptides. Certain formyl dipeptides and tripeptides containing methionine released histamine from human basophils at concentrations of 10(-4) to 10(-7) M. However, N-formyl amino acids did not release histamine. Tripeptides, in general, were more active than dipeptides. An acyl group was required for histamine release although an N-terminal position for Met was not essential. Histamine release from human basophils by these peptides correlated well with their chemotactic activity for rabbit leukocytes.


Recruitment of effector lymphocytes by initiator lymphocytes. Recruited lymphocytes are immunospecific and include graft-versus-host-reactive lymphocytes. We have shown previously that initiator T lymphocytes (ITL), sensitized in vitro against fibroblast antigens, recruit effector T cells in vivo. After injection into hind footpads of syngeneic recipients, sensitized ITL migrated to the draining popliteal lymph nodes (PLN) and activated a trapping mechanism by which circulating lymphocytes were recruited in the PLN. This paper reports experiments designed to test the immunospecificity of these recruited T lymphocytes (RTL). We found that immunospecific RTL were depleted from other lymphoid organs during recruitment in the PLN. However, immunospecific ITL were not depleted from spleens during PLN recruitment. Thus ITL and RTL are functionally distinguishable. We show that specific GVH reactive lymphocytes were also lost from spleens and distal lymph nodes during trapping of RTL in the PLN. Thus, the trapping phase of the recruitment response is immunospecific, as are the sensitization and effector phases. The trapped RTL are antigen-specific, and include the pool of GVH-reactive-lymphocytes committed to the same alloantigen. Thus, it appears that GVH-reactive cells respond to syngeneic ITL sensitized against allogeneic fibroblasts.


Immune response of the mouse to the major core protein (p30) of ecotropic leukemia viruses. Sera from normal C57BL/6 mice contained low titers of antibodies against proteins of MuLV. Sera from C57BL/6 mice that were immunized with allogeneic leukemia cells sometimes contained high-titered antibodies against the p15 protein of MuLV; these antibodies detected group-specific antigenic determinants of the p15 protein, since reactions were observed with the p15 proteins of both AKR and Moloney viruses. In contrast, antisera prepared in C57BL/6 mice against the AKR leukemia K36 reacted strongly with the p30 protein of MuLV, as well as with p15. Antibodies in the C57BL/6 anti-AKR K36 sera detected group-specific antigenic determinants of the p30 protein; reactions were observed with the C57BL/6 anti-AKR K36 serum and the p30 proteins of both AKR and Moloney viruses. It was concluded that mice do have the capacity to respond immunologically to antigenic determinants of the MuLV p30 protein, although in most circumstances this is not observed.


Defective release of eosinophil chemotactic factor from peripheral leukocytes in patients with chronic urticaria. Release of eosinophil chemotactic factor (ECF) and histamine was studied peripheral leukocytes of 16 normal and 12 ragweed allergic volunteers and compared to 15 patients with chronic urticaria. Cells were exposed to varying concentrations of anti-IgE or to ragweed antigen E in the case of allergic donors. Twelve of 15 patients with chronic urticaria showed defective release of ECF as well as histamine, while almost all normal and all allergic donors were able to release larger quantities of both mediators. Since the eosinophil is thought to have a modulating effect at sites of inflammation induced by anaphylactic mechanisms, the defective release of a factor attracting these cells may explain the persistence of symptoms in chronic urticaria.


Influenza virus subunit vaccines. II. Immunogenicity and original antigenic sin in humans. Subunit vaccines containing hemagglutinin, neuraminidase, and nucleocapsids of A/Port Chalmers/1/73 (H3N2) influenza virus were prepared after treatment of purified virus with ammonium deoxycholate. The immunogenicity of these subunits and the response to the common and specific antigenic determinants on the hemagglutinin subunits were studied in man. The subunits were as immunogenic in man as intact inactivated influenza virus vaccine at an equivalent concentration. Booster doses of antigen did not increase the antibody responses. Intact influenza B virus vaccine did not potentiate the immune response to the type A subunits in man. Volunteers responded differently to the common and specific determinants on the hemagglutinin subunits. The predominant antibody response was to the common or cross-reacting determinants present on hemagglutinins of both A/Hong Kong/1/68 (H3N2) and Port Chalmers/73 virus. Some men who failed to produce specific antibodies to the hemagglutinin of Port Chalmers/73 virus responded to the specific determinants on the Hong Kong/68 hemagglutinin. Higher doses of the subunit vaccines (1,400 chick cell-agglutinating units) did induce antibodies to the specific determinants on the Port Chalmers/73 virus hemagglutinin as well as to the common and specific determinants on Hong Kong/68 influenza virus.


Mechanism of escape, extrasystolic, and parasystolic arrhythmias. Study on an electrical analogue. A simple analogue of the heart consisting of a system of neon relaxation oscillators is presented. The analogue may display rhythm patterns similar to sinus rhythm, escape rhythm, isorrhythmic dissociation with synchronization, atrial extrasystoles, ventricular extrasystoles, and parasystole. The strict rules followed by these arrhythmias, as well as the deviations from the rules commonly followed by the equivalent heart arrhythmias, may be easily reproduced on the analogue. Such features are the Treppe phenomenon and captured beats in escape rhythm, fixed coupling intervals in extrasystoles, partial or complete atrioventricular block in very premature atrial extrasystoles, prolongation of the period following an atrial extrasystole, interpolated premature beats, complete compensatory pause and the rule of bigeminy in ventricular extrasystoles, slight instability of the parasystolic period, multiple length parasystolic periods slightly different from the exact multiples of the parasystolic idioperiod, preference of the parasystoles for certain phase in the sinus cycle, synchronization at a phase difference and fluctuation repeatedly and without interruption from a parasystolic to an extrasystolic rhythm and synchronization in escape rhythm with isorrhythmic dissociation. The mechanisms involved in these phenomena are discussed in detail. The striking similarity between the properties of the cardiac pacemakers and those of the relaxation oscillators on the one hand and betwen the rhythm patterns of the heart and those of the analogue on the other may permit the hypothesis that the mechanisms operating in the analogue may be used in analyzing and understanding heart arrhythmias.


Humoral immune responses in Rana catesbiana frogs and tadpoles. Rana catesbiana adult frogs and tadpoles were immunized with the bacteriophage F2, 0X-174, and T4 and the haptens 2,4 dinitrophenyl (DNP) and fluorescein (FTC). The haptens were conjugated with bovine serum albumin (BSA), bovine gamma globulin (BGG), or horsehoe crab hemocyanin (Hycn). Sera were obtained from immunized animals at invervals up to six months after immunization. The antibody activities were measured by bacteriophage neutralization techniques. Sucrose density gradients were used to separate the antibody classes. Both adults and tadpoles responded to each of the antigens tested. High molecular weight antibodies were predominant in both groups of animals. Low molecular weight antibody activity was not found in adults until nine weeks post immunization but, thereafter, this fraction increased throughout the immune response. Low molecular weight antibodies could also be identified in serum of tadpoles, but only under certain conditions.


Stepwise dissociation of high molecular weight avian myeloblastosis virus RNA: 30-40S RNA subunits--the best natural template-primer for viral reverse transcriptase. Controlled disruption of 60S AMV RNA with formamide was used to prepare 50-55S and 30-40S RNAS. When the activities of these RNAs as templates for AMV reverse transcriptase were compared it was found that 50-55S RNA was 1-5 times and 30-40S RNA 2 to 3 times more active than 60S RNA. The 30-40S RNA produced by heating, instead of formamide disruption, was inactive as a template but activity was restored by addition of oligo(dT). 40% of the 4S RNA initially associated with the 60S RNA remained associated with all the RNA species obtained by formamide treatment but was lost on heating. It is concluded that this RNA acts as resident primer whereas the other 60% of the 4S RNA is less firmly bound and appears to have little or no primer activity. Removal of the less firmly bound 4S RNA increases the template activity of the viral RNA.


Hepatitis B antigen: IgG components shown by immune electron microscopy. Immune complexes could be formed with hepatitis B Ag and anti-IgG serum after treatment of the antigen with detergent, CsCl or glycerol, but not before. It is suggested that an antigenic determinant specifically reacting with anti-IgG forms an integral part of hepatitis B Ag. This determinant is ordinarily obscured in the undamaged antigen.


Staining methods for an electron microscopic analysis of Golgi impregnated nervous tissue and a demonstration of the synaptic distribution upon pulvinar neurons. A Golgi impregnation method was modified for electron microscopy by reducing the time of the staining period. An acceptable level of the ultrastructural preservation was produced in several species of animals. Various methods were examined for removing the dense silver precipitate in order to show organelles in the stained neurons. Some methods for identifying particular parts of stained neurons in electron microscopy were also investigated by observing a sectioned block under an incident light microscope. The Golgi-EM method was applied to a study of the pulvinar nucleus in the rhesus monkey in order to demonstrate the usefulness and the limits of the method. Three neurons were studied, and at least four types of terminals (RL, F1, F2 and RS) were found synapsing on them. The distribution pattern of each type of terminal was determined upon the three neurons.


Visual pattern evoked responses and blink reflexes in assessment of MS diagnosis. A clinical study of 135 multiple sclerosis/pathol. VEPs were measured after pattern reversal in 135 MS patients and 30 control subjects. Neurological findings were documented in a standard manner. An extensive ophthalmological examination of all subjects was part of the study. The latency of P2 was abnormally delayed in 82% of the "definite", in 60% of the "probable" and in 65% of the "possible" MS groups respectively. The VEP was more often delayed in relation to ophthalmological disturbances. Changes in the MS classification had to be made in more than 10% of the patients, due to delay of VEP latency. These were patients with a "spinal" form of MS, which is known to create diagnostic problems. Optically and electrically evoked blink reflexes were recorded in 107 MS patients. All patients with mesencephalic lesions had delayed responses of the optically evoked reflex. 74% of the patients with caudal brainstem lesions had delayed latencies of the components of the electrically evoked blink reflex. The blink reflex was delayed in 18 additional patients without brainstem signs. The possibility of delineating clinically silent brainstem lesions by investigating blink reflexes is discussed.


Leucocyte migration inhibition in myasthenia gravis. The effect of thymectomy. Cellular hypersensitivity in myasthenia gravis (MG) to the thymus, muscles and peripheral nerves was examined by the method of the leucocyte migration inhibition test. The group of MG patients without thymoma had inhibition of leucocyte migration by thymus antigens. After thymectomy, they had a normal value of leucocyte migration. However, in the group of MG patients with thymoma, the inhibition of leucocyte migration by thymus antigens was observed after thymectomy. No significant inhibition of leucocyte migration was observed using muscle and peripheral nerve antigens. Cellular immunity in myasthenia gravis and the pathogenesis of the disease was discussed.


Measurement of diphenylhydantoin and phenobarbital by enzyme immunoassay and gas-liquid chromatography. A new enzyme-multiplied immunoassay technique (EMIT) was compared with Kupferberg's gas-liquid chromatography (GLC) for the determination of diphenylhydantoin and phenobarbital. 116 plasma samples of patients with epilepsy were examined simultaneously by EMIT and GLC. The precision of repeated determinations was similar for both procedures. There was good agreement between the EMIT and GLC results. The rapid and precise analysis, made possible by the EMIT system, is useful for the treatment of epilepsies.


The fatty acid composition of major glycosphingolipids (cerebrosides and sulfatides) in human cerebral white matter measured by a simple micromethod. A micromethod for the investigation of the fatty acid composition of myelin glycosphingolipids (cerebrosides and sulfatides) suitable for general application in the investigation of neurological disorders, especially demyelinating diseases, is presented. Using the lipids extracted from 1 g of material these are freed of phospholipids by Florisil column chromatography and separated by thin-layer chromatography into 2 cerebroside and sulfatide fractions which are analyzed individually. The results obtained from the white matter of 13 normal adult brains are distributed within a narrow range which is most pronounced for the group of long chain fatty acids. Our results also agree with those quoted from literature.


[Neurological and psychiatric disorders following acute arsine poisoning (author's transl)]. Follow-up study of 6 workers, who after survival of an acute arsine poisoning, developed psychopathologic and neurologic abnormalities. The symptoms appeared after a latency of 1 to 6 months indicating a toxic polyneuropathy and a mild psycho-organic syndrome. The severity of these reversible manifestations was directly related to the period of time of exposure to arsine. The clinical picture of arsine polyneuropathy was similar to that observed in arsenic poisoning, suggesting that arsine polyneuropathy is due to the action of arsenic. The psychopathologic syndrome corresponds to the so-called "Vergiftungsspätfolgesyndrom" and therefore does not appear to be a specific sequel of arsine poisoning.


Pineal apoplexy associated with anticoagulant therapy. Case report. A case is presented with the acute development of a pineal region syndrome secondary to hemorrhage into a pineal cyst in a patient who was under anticoagulant therapy. Resolution of the symptoms and signs followed excision of the lesion.


Mental health nursing consultation: a study of expectations. Psychiatric nurses working in the field of mental health consultation to medical-surgical nursing units may find it useful to become more fully aware of what is expected of the mental health nurse consultant role and how they can best meet these expectations in their individual situations and setting. This study demonstrated that the medical-surgical nurses in one hospital do have specific expectations for the role of the mental health nurse consultant. To fulfill these expectations the mental health nurse consultant should identify her role in light of the above findings and attempt to integrate her job within the boundaries set forth by her hospital, the nursing team with whom she is working, and the patient's needs.


Histamine release by exocytosis from rat mast cells on reduction of extracellular sodium: a secretory response inhibited by calcium, strontium, barium or magnesium. 1. Histamine release from peritoneal mast cells of the rat was stimulated when the cells were exposed for 10 min to sodium-deficient media where all NaCl had been replaced by KC1, RbC1, glucose, sucrose, mannitol, or Tris, provided calcium was less than about 0-5 mM. 2. Light and electron microscopy showed the response to be exocytosis. 3. The chelating agents, EDTA and EGTA, abolished the response to sodium lack and their inhibitory effects were reversed by re-incubating cells with calcium but not magnesium. 4. The response was inhibited by dinitrophenol combined with glucose-deprivation. 5. The response was inversely related to the concentrations of sodium and calcium below 137-5 and 0-5 mM respectively. 6. The related alkaline earth metals, barium, strontium, and magnesium, resembled calcium in inhibiting the response to sodium lack. 7. No secretory response was seen when the cells were exposed for 10 min to calcium-free medium in which lithium replaced sodium. Exposure to this medium for 60 min, however, elicited secretion. 8. It is concluded that when extracellular calcium is low, a reduction in extracellular sodium induces a conventional exocytotic secretory response dependent on energy and cellular calcium. It is suggested that sodium lack may mobilize calcium from a cellular site possibly the inner aspect of the plasma membrane.


The effect of nursing intervention on a program of behavior modification by parents in the home. The purpose of the study was to determine if nursing intervention affects a program of behavior modification for neurologically impaired children carried out by parents in the home. Nine neurologically impaired subjects between the ages of two and four years were selected for programs of behavior modification by their parents at home. The nine subjects were randomly divided into an experimental and control group. The groups were generally matched for age and socioeconomic status. The four subjects in the experimental group were visited bi-weekly for a two-month period after the initial program was set up with changes, assessments and/or alterations made in the programs as necessary. The five subjects in the control group were visited at the beginning of the study to establish the programs of behavior modification the parents would implement. This group was not revisited until the end of the two-month period. The findings indicated a marked change in the subjects of the experimental group. This difference was significant on the basis of Wilcoxin Matched Pairs p. greater than .00003. In the control group only one child had any appreciable change in the specific target behavior, and it was not the desired goal.


Observations on the production of frozen-dried thin sections for electron microscopy using unfixed fresh liver, fast-frozen without cryoprotectants. Thin sections of unfixed liver, fast-frozen without cryoprotectants, have been cut using conditions under which momentary thawing of the sections is unlikely to occur. A transfer stage which facilitates this procedure is described. Sections show hole damage probably due to ice-crystal formation during the freezing process and have well defined edges, but despite hole damage, some morphological features of the cell are discernible. Presumptive mitochondria appear smaller in frozen sections than in conventional Araldite sections. Sections devoid of hole damage have indistinct edges and are presumed to have undergone transient thawing. Carbon coating of freeze-dried sections to exclude atmospheric moisture during transference of sections to the electron microscope (EM) appear unnecessary as regards preservation of morphological structure. The results are discussed in relation to the limitations of the method and the potential value of the technique.


Serum-soluble malarial antigens and immune complex nephritis in Plasmodium berghei berghei infected mice. Swiss albino mice infected with Plasmodium berghei berghi showed the serum-soluble malarial antigen and antibody on day 10 of infection onward. Immune complex nephritis in these mice developed on the seventh day after inoculation. The infected kidneys revealed the deposition of mouse gamma globulin, mouse beta1C globulin and malaria antigen along the capillary wall of the glomeruli. Proteinuria was detected on seventh day of infection. Serum-soluble malaria antigen in probably responsible for forming the soluble immune complex which causes glomerulonephritis in infected mice.


Early synthesis of Semliki Forest virus-specific proteins in infected chicken cells. Cells preinfected with fowl plague virus followed by treatment with actinomycin D are a suitable system for studying early protein synthesis in cells infected with Semliki forest virus. One and one-half hours after superinfection, three new nonstructural proteins (NVP) were detected: NVP 145, NVP, 112, and NVP 65. They appeared in parallel with a low incorporation of mannose at the beginning of the infectious cycle. Behavior on chasing suggested a precursor relationship of NVP 112 to the envelope glycoproteins. Two kinds of NVP 65 are described, both of which are varieties of NVP 68 with an incomplete mannose content. One type, detected early after infection, was converted into NVP 68 by supplementary glycosylation. The second, late type was stable. It contains fucose and resembles the NVP 65 observed after impairment of glycosylation. The mechanism of NVP 68 glycosylation is discussed. The presence of the complete carbohydrate moiety is crucial for the cleavage of NVP 68 into the envelope proteins E2 and E3 and, thus, for virus maturation. Only the complete form of NVP 68 was precipitated by envelope-specific antisera. A large production of NVP 78 is a further feature of the early events in infected cells. It is not related to the structural proteins.


Effect of passage in culture on a clone of BALB/c 3T3 cells transformed by simian virus 40. Most simian virus 40 (SV40)-transformed BALB/c 3T3 clones employed for biochemical studies have been used without regard to passage level. To determine whether virus-induced properties are stable as a function of passage, we have extensively characterized one transformed clone, FNE, which was isolated after SV40 infection BALB/c 3T3 cells in factor-free medium. From the initial testing at passage 5 and for at least 50 subsequent subcultures, the cells stably maintained many transformed growth properties, including high saturation density, morphology, colony formation on contact-inhibited monolayers, tumorigenicity, and synthesis of viral-specific RNA. However, other properties varied as a function of passage. There was a slight decrease in viral genome equivalents per cell from 1.1 copy/cell at passage 5 to 0.7 copies at passage 40. Initially, the cells were negative for all type C virus; however, cells carried at low density for 13 to 20 passages (65 to 100 generations) began to release an endogenous type C virus that then persisted in the culture. Spontaneous release of type C virus did not occur in control BALB/c 3T3 cells carried under identical culture conditions for 90 passages. When the cultures were releasing type C viruses they stained uniformly and brightly positive for SV40 tumor (T) antigen by immunofluorescence, whereas T antigen staining was variable at early passage. These experiments suggest that subtle but perhaps important differences in viral gene expression can occur as a function of passage; they also demonstrate the importance of evaluating the interactions between SV40 and endogenous type C viruses.


Naturally occurring murine leukemia viruses in wild mice: characterization of a new "amphotropic" class. A new class of murine leukemia viruses, isolated from wild Mus musculus trapped in California, is described. These viruses, designated "amphotropic," replicate in mouse, rabbit, mink, human, guinea pig, and rat cells, but not in hamster, quail, or duck cells. They show N-tropism for mouse cells, and do not trigger the XC cell response. They are distinct by interference and virus neutralization testing from the previously recognized mouse-tropic and xenotropic MuLV classes. Mouse-tropic viruses occuring along with three of the four amphotropic isolates were found to be distinguishable by virus neutralization from other mouse-tropic murine leukemia virus strains of laboratory mouse origin.


Sequence homology between Moloney murine sarcoma virus and Moloney leukemia virus RNA. The Moloney murine sarcoma-leukemia virus [M-MSV (MuLV)], propagated at high multiplicity of infection (MOI), was demonstrated previously to contain a native genome mass of 4 X 10(6) daltons as contrasted to a mass of 7 X 10(6) daltons for Moloney murine leukemia virus (M-MuLV). The 4 X 10(6)-dalton classof RNA from M-MSV (MuLV) was examined for base sequence homology with DNA complementary to the 7 X 10(6)-dalton M-MuLV RNA genome. Approximately 86% of the M-MSV (MuLV) was protected from RNase digestion by hybridization, whereas 95% of M-MuLV was protected under identical conditions. These results indicate that the small RNA class of high-MOI M-MSV (MuLV) contains little (perhaps 10%) genetic information not present in M-MuLV. Virtually all of the 1.8 X 10(6)-dalton subunits of M-MSV (MuLV) RNA contained regions of poly(A) since 94% of the RNA bound to oligo(dT) cellulose in 0.5 M KCl. This suggests that the formation of the 1.8 X 10(6)-dalton subunits occurs before their packaging into virions and does not result from hydrolysis of intact 3.5 X 10(6)-dalton subunits by a virion-associated nuclease.


The value of palliative spinal surgery in metastatic urogenital tumors. There were 22 patients with vertebral metastases and impending neurologic damage from prostatic, vesical and renal cell carcinoma treated by decompressive laminectomy combined with local radiotherapy and chemotherapy. The neurological recovery was complete in the prostatic carcinoma patients (10 of 10), partial in the vesical carcinoma patients (2 of 5) and unsatisfactory in the renal cell carcinoma patients (1 of 7).


Phyllodes type of atypical prostatic hyperplasia. A prostatic lesion characterized by atypical epithelial hyperplasia and pleomorphism of the stromal elements is presented. The features bear a strong resemblance to cystosarcoma phyllodes of the female breast and probably represent a comparable lesion. Such atypical hyperplasia has not been described previously.


Analgesic cystitis: an unusual microangiopathy of the bladder. A brownish discoloration of the bladder mucosa around the ureteral orifices has been observed on cystoscopic examination in 3 patients taking excessive quantities of analgesics. Biopsy revealed an unusual degree of hyalinization of the capillary walls in the submucosa.


Accuracy of preoperative estimation of resection weight in transurethral prostatectomy. In a prospective study of 150 consecutive transurethral resections of the prostate gland the preoperative estimate of gland weight was compared to the actual resected weight. The data revealed a wide variation between estimated and actual weight in all size glands.


Effect of bacterial lipopolysaccharides on anti-trinitrophenyl antibody-producing cells. Nonspecific modification of the affinity at the cellular level. The effect of lipopolysaccharide (LPS) on anti-trinitrophenyl (TNP) direct plaque-forming cells (PFC) in the spleen of mice and the affinity of antibodies produced by these PFC were examined. Simultaneous injection of bacterial LPS and TNP-coupled sheep red blood cells(SRBC) induced an obvious increase in anti-TNP PFC numbers and heightened the antibody affinity at cellular levels. The higher the doses of LPS, the greater the effects. Concomitant injection of LPS in TNP-coupled homologous mouse red blood cells (MRBC) also elicited good anti-TNP PFC response and slightly heightened the affinity. Priming with LPS and SRBC together 7 days prior to immunization did not enhance the anti-TNP PFC response and it was difficult to alter the affinity. Preinjection with small amounts of TNP-MRBC or -rabbit red blood cells and LPS simultaneously did not induce any significant increase in anti-TNP PFC secondary response after reimmunization with TNP-SRBC, but obviously heightened the antibody affinity. Injection of LPS simultaneously with the secondary immunization was effective for both the anti-TNP PFC response and the alteration of antibody affinity. These results suggest that LPS affects the control mechanisms of anti-TNP antibody affinity via the non-thymus-derived helper cell function, and the adjuvant action and alteration of antibody affinity induced by LPS are regulated by different mechanisms.


Interaction of human Waldenström's IgM proteins with guinea pig and human complement. The activity of purified human Waldenström's IgM protein to fix complement of human and guinea pig origins was compared at different temperatures using the polystyrene latex particle-adsorption method. It was shown that the interaction of the IgM proteins with complement differed depending on the source of complement and that a pronounced heterogeneity in complement-fixing activity was observed among the IgM proteins when tested with guinea pig complement. Thus, by the use of guinea pig complement, six human IgM proteins examined were classified roughly into two groups, one having a high and the other a low activity at 3 C as well as at 37 degrees C. With human complement, five proteins showed a rather uniform activity at 37 degrees C. However, there was one protein with no detectable activity, suggesting the presence of non-complement-fixing protein in the IgM class. All the six proteins showed no significant activity with human complement at 3 C. No antigenic difference has been found as yet in the Fc or Cmu2 region among these IgM proteins examined.


Immunochemical studies on bacterial blood group substances. VIII. The structure of oligosaccharides from blood group H-active polysaccharide of Escherichia coli 2B-V. Blood group H-active  polysaccharide has been prepared from "smooth" strain Escherichia coli 2B-V by Freeman's method, alpha-Fucosidase derived from Bacillus fluminans caused the liberation of fucose from this polysaccharide, together with concomitant loss of blood group H activity. The results of quantitative microanalysis, borohydride reduction, the Morgan-Elson reaction and enzymic hydrolysis with betagalactosidase using isolated oligosaccharides obtained by partial acid hydrolysis indicated that the O-specific side chain of the polysaccharide has a pentassaccharide unit which is beta-D-Gal-(1 leads to 3)-D-GalNAc-(1 leads to 3)-D-GalNAc-Fuc with a D-glucose residue bound at some undetermined point on this structure. It was considered that terminal non-reducing fucose of the polysaccharide was liberated by partial acid hydrolysis.


[Diagnostic importance of visken -- an adrenergic beta receptor blocking agent]. Among the newest beta-blocking agents visken is the one that either produces no or very little adverse inotropic effect. In some groups of patients with organic and neurotic cardiac disorders, screened as a result of a careful clinical examination, changes that occurred on the ECG with 12 leads following an intake of a single visken tablet (5 mg) were studied. Ingestion of visken was not seen to bring any improvement in the ECG findings in 88 per cent of 50 patients with organic disorders, most of whom had sclerosis of the coronary arteries and hypertensive disease attended by a high diastolic pressure. A thorough clinical examination of 70 patients presenting neurotic conditions helped to rule out any organic cardiac lesions, and in them the diastolic pressure did not exceed 95 mm Hg. The visken test showed that in 91 per cent of these cases the ECG returned back to normal and only in 1.5 per cent of them no changes could be observed. In both of the above groups the effect of visken was statistically significant (p less than 0.001). Repolarization derangements in the I and II leads among patients with organic lesions was observed 4 times as often as in the II and III leads. Among neurotic patients changes recorded in the II and III leads were twice as frequent as in the I and II leads.


Imitation: a closer look.

Imitation: a closer look. This paper explores the effectiveness of elicited imitation in clinical assessment and remediation procedures. A critical review of the literature dealing with the role of elicited imitation in language acquisition as well as the use of elicited imitation clinically is presented. Conclusions drawn from the review indicated that the effectiveness of employing elicited imitation for assessment and remediation purposes is not clearly defined.


RNA tumor virus-like activities in human solid tissues: endogenous RNA:DNA polymerase activities in the prostate. Several human prostatic tissues have been examined for possible particles and associated DNA polymerizing activity generally associated with the C-type RNA tumor virus family. Partially purified tissue extracts, when centrifuged to equilibrium in sucrose gradients, yield fractions which contain actinomycin D resistant, endogenous DNA polymerase activity; this activity bands at a density of 1.15-1.18 gm/cm3. Further analysis of the endogenous products by sucrose gradient sedimentation suggested the presence of high molecular weight RNA:DNA hybrids generally felt to be indicative of a faithful copy of a lengthy stretch of viral specific RNA. However, most of the DNA products synthesized in these endogenous reactions sedimented in much lower molecular weight regions of these sucrose gradients. Clearly, the relative distributions of "high" and "low" molecular weight products could critically depend on the nuclease content of the subcellular fraction under study, and the prostate may be relatively enriched in nucleases. Further, oligo (dT) stimulated the endogenous DNA polymerase activity contained in these extracts, and omission of one of the DNA precursor nucleotides depressed it. Thus, it seems unlikely that terminal transferase activity, rather than genuine DNA polymerization, was being measured primarily. Because of the spectrum of molecular weight classes formed by these DNA:RNA hybrids, as well as their apparent presence in normal prostatic tissue, we find it difficult to ascribe their presence with certainty either to the presence of typical C-type RNA viruses or to the exclusive behavior of the neoplastic prostatic tissue. Thus, our studies lend support to the growing evidence for functions similar to those of C-type RNA viruses being relatively widespread in human tissues without the apparent necessity for a possible etiologic role in neoplastic production (Strand and August, 1974; Sherr et al., 1974). At the same time, our current studies emphasize the need for caution in drawing conclusions from results utilizing probes generally felt quite useful in scoring for presence of virus in lower animals at least in the human prostate.


[Juvenile hyalin fibromatosis].

[Juvenile hyalin fibromatosis]. The focal lesions of the skin and mucosae of this rare disease, that has been described as juvenile hyalin fibromatosis, are initially in moderately elevated plaques, that evolve to form nodules in the dermis. The fundamental defect, possibly of autosomal recessive inheritance, is a disturbance in the metabolism of the mucosaccharides. There is an accumulation, both intracellularly, in membrane-limited vacuoles, and extra-cellulary, of fibrilar glycoprotein complexes, the saccharides of which are both neutral and acidic, non-sulfated and sulfated. The material accumulated in the interstitium has hyalin aspect and is poorer in sulfated mucosaccharides than the intracellular vacuoles. The defect manifests itself primarily in the mesenchymal perviascular cells, leading to their proliferation.


[Determination of B lymphocytes: immunofluorescence and peroxidase dyes (preliminary note)]. A determination of the lymphocyte levels in blood and cutaneous infiltrates of 60 patients complaining of different dermatoses were carried out by the authors. The method used in the experience was that of Preud'homme and Flandrin (1974). In most of the investigated cases the results could be considered as normal (8-22%). In one case of actinic reticuloid and 2 cases of plane xanthoma with photosensitivity the levels were considerably raised. The authors belleve that the B-lymphocytes determination could be worth investigation in the photosensitive dermatoses.


Malignant histiocytic reticulosis. Effect of bleomycin by intra-arterial route. Report of a case. A case of malignant histiocytary lymphoma treated with Bleomycyn in a 78 year old patient is reported. The results were good but not durable.


An outbreak of hepatic veno-occlusive disease in north-western Afghanistan. Following a 2-year period of severe drought a very large number of patients with massive ascites and emaciation were observed in north-western Afghanistan. Clinicopathological study showed that these were typical cases of hepatic veno-occlusive disease. The outbreak was caused by consumption of bread made from wheat contiminated with seeds of Heliotropium plants, which were shown to contain pyrrolizidine alkaloids. Examination of 7200 inhabitants from the affected villages showed evidence of liver disease in 22.6%. Clinical improvement was observed in thirteen cases after 3 to 9 months of supportive hospital treatment, and in three cases liver biopsies showed almost complete disappearance of initial abnormalities.


An epidemic of veno-occlusive disease of liver in central India. An outbreak of veno-occlusive disease which was probably caused by consumption of cereals mixed with seeds of a plant (Crotalaria sp.) containing pyrrolizidine alkaloids occurred in the Sarguja district of India in November-December 1975. 42% of the 67 recorded cases died.


Persistent gastrointestinal bleeding due to angiodysplasia of the gut in von Willebrand's disease. Gastronintestinal vascular dysplasia as a cause of persistently recurring melaena is reported in two patients with von Willebrand's disease. Colonic wall vasculature was affected in one patient, the abnormality being demonstrated both radiologically and at operative hemicolectomy undertaken to control bleeding. Abnormalities in the vasculature of the duodenal and jejunal walls were found in the second patient at operation and necropsy, though not demonstrated by angiography.


Bromocriptine and levodopa (with or without carbidopa) in parkinsonism. Bromocriptine, a dopamine receptor agonist, was administered to 20 patients with idiopathic parkinsonism taking levodopa (L-dopa) or "Sinemet" (levodopa combined with carbidopa in a 10/1 ratio) at optimum doses. In a double-blind randomised cross-over study lasting 6 months, the addition of bromocriptine (mean daily dose 79 mg) led to a significant (P less than 0.01) 74% reduction in the dose of sinemet and levodopa. "Total disability score" showed a significant (P less than 0.01) improvement at both low and high doses of bromocriptine. Tremor improved 50% (P less than 0.01), with significant improvements in gait, posture, writing, balance, rigidity, finger dexterity, and drooling. Adverse reactions were similar to those observed with sinemet and levodopa. Although both the cause and the cure of idiopathic parkinsonism remain elusive, bromocriptine appears to represent a therapeutic advance.