Protein allergens of white-faced hornet, yellow hornet, and yellow jacket venoms. White-faced hornet, yellow hornet, and yellow jacket venoms have very similar protein compositions; each contains mainly three basic proteins. Two of these proteins have hyaluronidase and phospholipase activities and the third one, designated as antigen 5, is of as yet unidentified biochemical function. These three proteins have molecular weights of about 45 000, 35 000, and 25 000, respectively. The three proteins of white-faced hornet venom have been purified to near homogeneity, while this is the case only for antigen 5 of yellow hornet and yellow jacket venoms. Strong antigenic cross-reaction of the hyaluronidase from these three vespid venoms was observed using specific rabbit anti-venom sera, while weak cross-reactions of phospholipases and of antigen 5s were observed. All three proteins are active as allergens to varying degrees in vespid sensitive individuals. With each vespid venom its antigen 5 seems to be the major allergen. The results help to clarify the commonly observed varying degrees of multiple sensitivity of people to different vespids.


Isolation of a porcine liver plasma membrane fraction that binds low density lipoproteins. Large amounts of injected radiolabeled low density lipoproteins have been found by others to accumulate primarily in the liver and studies in various types of isolated cells, including hepatocytes, have indicated the presence of specific cell membrane recognition sites for lipoproteins. In the present studies, the high affinity binding of radiolabeled low density lipoproteins ([125I]LDL, d 1.020--1.063 g/mL) was measured in the major subcellular fractions of porcine liver homogenates. The nuclear and mitochondrial fractions were 1.9- and 1.4-fold enriched in binding activity with respect to unfractionated homogenates and contained 15% and 12% of the total binding activity, respectively. The  microsomes, which contained most of the plasma membranes and endoplasmic reticulum, were approximately 4-fold enriched in binding and contained 73% of the binding activity. Microsomal subfractions obtained by differential homogenization and centrifugation procedures were 5.6--7.0-fold enriched in LDL binding and contained 54--58% of the homogenate binding activity. They were separated by discontinuous sucrose density gradient centrifugation into fractions which contained "light" and "heavy" plasma membranes and endoplasmic reticulum. The heavy membrane fraction was 2--4 fold in binding with respect to the parent microsomes (16--22 fold with respect to the homogenate). There was no enrichment of binding activity in the other two fractions. Two plasma membrane "marker" enzymes, nucleotide pyrophosphatase and 5'-nucleotidase, were also followed. Of the two, binding in the sucrose density gradient subfractions most closely followed nucleotide pyrophosphatase, which was also most highly enriched (3.2--3.3-fold) in the heavy membrane fraction, but did not follow it exactly. The enzyme was 2-fold richer in the light membranes than in the parent microsomes, though the light membrane binding activity was only 0.4--1.4 times that of the parent microsomes. High affinity binding was time and temperature dependent, saturable, and inhibited by unlabeled low density lipoproteins but not by unrelated proteins. Binding was stimulated 2--3 fold Ca2+, was not affected by treatment with Pronase or trypsin and was inhibited by low concentrations of phospholipids and high density lipoproteins (HDL). Heparin-Mn2+ treatment of HDL did not affect its ability to inhibit [125I] LDL binding. The LDL recognition site was distinct from the liver membrane asialoglycoprotein receptor; LDL binding was not inhibited by desialidated fetuin. We conclude that porcine liver contains a high affinity binding site that recognizes features common to both pig low density and high density lipoproteins. Further studies may elucidate the significance of this binding site in lipoprotein metabolism.


Hormonal induction of alpha2u-globulin synthesis in isolated rat hepatocytes. Hepatocytes freshly prepared with collagenase synthesize alpha 2u-globulin and other hepatic proteins in vitro at approximately the same rate throughout 30 h of incubation. The newly synthesized proteins are efficiently secreted into the medium throughout this period. That the secretion of proteins by hepatocytes is not due to cell leakage is shown by the fact that 30 micrometer colchicine prevents the appearance of labeled alpha2u-globulin and other proteins in the medium. Hepatocytes isolated from animals in different endocrine states synthesize alpha2u-globulin in vitro at rates consistent with the hormonal effects upon its in vivo biosynthesis. In vitro addition of androgens to hepatocytes isolated from castrated male rats evokes an elevation of general protein synthesis in these hepatocytes. Glucocorticoids, added in vitro, specifically induce and elevated rate of alpha2u-globulin synthesis.


Location of major antigenic sites of the beta subunit of human chorionic gonadotropin. Purified beta subunit of human chorionic gonadotropin (hCG) was partially reduced with beta-mercaptoethanol, carboxymethylated, and digested with chymotrypsin. The peptides were isolated by high-voltage electrophoresis and paper chromatography. Five major disulfide-containing peptides were isolated, and their location in the parent molecule was established by amino acid composition and amino- and carboxy-terminal analyses. All of these peptides inhibited the binding of 125I-labeled hCG by anti-beta hCG serum. The inhibitory effect of these peptides was lost when their disulfide bonds were reduced and alkylated. Synthetic carboxy-terminal peptides were not inhibitory. Based on these data it is concluded that a major antigenic site of hCG resides in the region of residues 21-23 with a disulfide bond connecting cysteine-23 or -26 with the cysteines at positions 72 or 110.


Binding of fatty acids and tryptophan to alpha-fetoprotein from fetal pigs. Using gel electrophoresis and autoradiography, we have shown that plamitic, stearic, oleic and arachidonic acids as well as tryptophan bind to alpha-fetoprotein derived from fetal swine serum. It is also shown that these ligands bind to albumin from both fetal and adult swine serum. The results suggest that alpha-fetoprotein in the fetus has transport functions similar to albumin in the adult.


Lipopolysaccharide interferes with the staining of lipoprotein on polyacrylamide gels. After electrophoresis of total membrane preparations of Escherichia coli B on sodium dodecyl sulfate polyacrylamide gels, and subsequent staining with Coomassie Brilliant blue, a band corresponding to the Braun lipoprotein fails to appear. This is in contrast to similar preparations of E. coli K-12 which do display the lipoprotein upon staining. Experiments described below indicate that failure to observe this protein in E. coli B is due to interference in the staining reaction by the lipopolysaccharide present in the membrane preparations.


The effect of antibiotics on the photocycle and protoncycle of purple membrane suspensions. The interrelation was studied between the phototransient absorbing maximally at 412 nm (M412) and light-induced proton release under steady-state conditions in aqueous suspensions of 'purple membrane' derived from Halobacterium halobium. The decay of M412 was slowed down by the simultaneous application of the ionophoric antibiotics valinomycin and beauvericin. The former had only slight activity alone and the latter was effective only in conjunction with valinomycin. The steady-state concentration of M412 which was formed on illumination was a direct function of the concentration of valinomycin. Maximum stabilization of M412 was obtained when the valinomycin was approximately equimolar with the bacteriorhodopsin. Addition of salts to the medium increased the number of protons released per molecule of M412 without affecting the level of M412 which was produced by continuous illumination. The effectiveness of the salts in this respect depended on the nature of the cation. Ca2+ and their antagonists La3+ and ruthenium red were found to have especially high affinity for the system. The extent of light-induced acidification could not be enhanced by increasing the pH of the medium from 6.5 to 7.8. The possible mechanism of action of the ionophores and of the cations on the photocycle and on the proton cycle is discussed.


Interactions in vitro and in vivo between rat serum protease inhibitors and anodal and cathodal rat trypsin and chymotrypsin. Reaction mixtures of increasing amounts of the pancreatic homologous proteases, anodal and cathodal chymotrypsin and trypsin, respectively, and normal rat serum were analyzed by immunoelectrophoretic methods in order to determine their distribution on serum protease inhibitors. This paper concerns three proteins occurring in normal serum and capable of binding protease viz. alpha1-macroglobulin, alpha1-antitrypsin and alpha1-inhibitor 3. The distribution of the enzymes among these protease inhibitors differed significantly from one protease to another. The distribution of the proteases among the serum protease inhibitors following intravenous injection of 125I-labelled proteases corresponded to that in vitro. Complexes formed with alpha1-macroglobulin and alpha1-inhibitor 3 were quickly eliminated irrespective of the enzyme species used, whereas those formed with alpha1-antitrypsin persisted much longer in the circulation.


Purification of tadpole collagenase and characterization using collagen and synthetic substrates. Tadpole collagenase hydrolyzed native and denatured collagen and synthetic peptides with sequences of 2,4-dinitrophenyl-L-prolyl-L-leucylglycyl-L-isoleucyl-L-alanylglycyl-L-arginie amide and 2,4-dinitrophenyl-L-prolyl-L-glutaminyl-glycyl-L-isoleucyl-L-alanylglycyl-L-glutaminyl-D-arginine. The specific enzyme activity against the latter substrate and collagen fibrils is found to be 933 nmol/min per mg protein and 8440 units (microgram collagen degraded/min), respectively. Optimum pH for the enzyme is 7.5-8.5. A collagenase complex with alpha2-macroglobulin did not hydrolyze collagen fibrils, but digested the synthetic substrates at the Gly-Ile bond. The amino acid composition of the enzyme was determined. Immunoelectrophoresis of the enzyme at pH 8.6 against anti-tadpole collagenase rabbit immunoglobulin G shows a single precipitin line at a position migrating faster than human serum albumin and corresponding to enzyme activity against collagen fibril and synthetic substrates.


[Electron paramagnetic resonance study of the interactions between steroid hormones and binding proteins]. Interaction of a spin labeled corticosteroid (desoxycorticosterone nitroxyde: DOC -NO) with three purified proteins (albumin, transcortin, progesterone binding protein: PBG) was studied by electron spin resonance (ESR) spectroscopy. DOC-NO was competitive with natural corticosteroids and therefore bound at the same site to specific binding proteins. ESR spectra in the presence of each of the proteins showed an immobilized (bound) form of the spin labeled steroid and allowed the calculation of the corresponding association constant (Ka) at equilibrium. The three binding proteins could be characterized by the ESR parameters of the DOC-NO bound form. The thermodynamic parameters (deltaH, deltaS) of the steroid-protein interactions were calculated from the ESR data obtained within a wide temperature range (3--40 degrees C). The ESR spectra width (2T) was used to evaluate the polarity of the spin label environment within the steroid binding site: a hydrophobic character was observed for transcortin whereas PBG exhibited a more hydrophilic steroid binding sits. The rotational correlation time of the three protein DOC-NO complexes at equilibrium were calculated from ESR data; the results were correlated with the protein molecular size and suggested a non spherical shape for the binding macromolecule in solution. Spin labelling of biologically active steroids thus provides a novel approach for the study of the interaction of these hormones with their binding protein. Providing a suitable spin label, the ESR parameters may allow the characterization of several types of binding sites of different biological significance for the same hormone, in biological fluids as well as in target tissues.


The biology of social attachments: opiates alleviate separation distress. The possibility that brain opiate systems participate in the control of social affect was assessed by determining capacity of low doses of exogenous opiates (0.125-0.50 mg/kg oxymorphone, and 0.10-0.50 mg/kg morphine sulfate) to reduce distress vocalizations of socially isolated puppies. Low doses of opiates were capable of profoundly reducing crying as well as the motor agitation they exhibit during brief periods of social isolation. Since reductions in crying could be obtained with morphine in the absence of any gross behavioral disturbances, the possibility is entertained that brain opiates may function to control the intensity of emotions arising from social separation. Possible parallels between the biological nature of narcotic addiction and the formation of social bonds are discussed.


[Cytophotometry of DNA in serial sections]. DNA of leprosy cells in serial paraffine sections stained after Felgen were studied photometrically in scanning microscope SPM with electron computer Wang 720C. Significant differences in extinction values (up to 30--35%) between individual sections within the same series were revealed. Cumulative results of different series were rather uniform. It was concluded that photometric studies of serial sections should be carried out for reliable determination of DNA.


Purification and characterization of mouse brain Thy-1.2 differentiation alloantigen. Subcellular fractionation of C57BI/6J mouse brains produced a crude synaptosome preparation which contained virtually all of the Thy-1.2 antigenic activity of the isotonic whole brain homogenate. The Thy-1.2 was solubilized from the synaptosomes, following delipidation with acetone, by deoxycholate extraction. A glycoprotein fraction rich in Thy-1.2 was isolated from the bulk of the detergent-soluble material by lectin affinity chromatography. Fractionation of the lectin retentate by gel filtration chromatography produced a single peak of Thy-1.2 activity purified more than 2000-fold over the original homogenate. SDS polyacrylamide gel electrophoresis of this material revealed a single band which corresponded to an apparent molecular weight of 24,000. Amino acid composition data indicated that the protein portion of the molecule is similar to Thy-1.1 from mouse lymphoblastoid cells. Carbohydrate analysis revealed a qualitative similarity between mouse brain Thy-1.2 and Thy-1.1 from rat brain. Structural differences which could account for the Thy-1.1 and Thy-1.2 antigenic distinctions are apparently too subtle to be detected by compositional analysis.


Comparison of active zones in spinal cord of anesthetized and unanesthetized rats: a high voltage electron microscope study. High voltage electronmicroscopy was applied to the study of presynaptic nerve terminals and specifically the arrangement and staining properties of synaptic vesicles in and around the active zone. The zinc iodideosmium tetroxide technique was used for the impregnation of neuropil derived from the spinal cord of anesthetized (40 mg/kg pentobarbital sodium) and unanesthetized rats. Under optimal conditions of section thickness and tilt angles stereoscopic views of a paracrystalline lattice arrangement of synaptic vesicles could be ascertained. The ZIO affinity was significantly higher in synaptic vesicles of the anesthetized as compared with the unanesthetized preparations.


Subcellular and polypeptide distributions of slowly transported proteins in the garfish olfactory nerve. In the garfish olfactory nerve proteins labeled with [3H]leucine are transported by slow axonal flow as a well-defined crest of radioactivity. At 21 degrees C slow flow moves along the axon with a velocity of 0.92 +/- 0.02 mm/day. It has been possible to analyze 4 subcellular fractions (soluble, mitochondrial and 2 membranous) as well as their polypeptide composition, in areas of the nerve containing (1) the slow moving crest, (2) the material remaining in the nerve behind the crest, and (3) the labeling present in front of the slow crest. Analyses were done 70 and 110 days after isotope deposition. The crest of slow moving radioactivity is characterized by a close parallelism between labeling and protein concentration in the subcellular fractions as well as among the polypeptides constituting these fractions. The radioactivity is mainly associated with mol. wt. of 14,000, 30-45,000, 58,000 and 68,000. This last peak corresponds to a protein not labeled by fast transport, present only in the light membranous fraction. The composition of the moving crest remains essentially constant during the 40-day period investigated. Most of the slow-moving molecules remain in the axon behind the moving crest. This deposited material appears to be redistributed and/or to be turning over more rapidly than the molecules still moving in the crest. A large amount of radioactivity was recovered in front of the moving crest. This might be produced by molecules deposited by fast transport and by material released from the cell body at rates intermediate between the fast and slow phases of transport. The subcellular and polypeptide compositions of this area of the nerve remain constant and are intermediate between the compositions of fast and slow flow. The slowly transported labeled polypeptides in the mitochondrial fraction are of low molecular weight, and were found to be similar in the various areas of the nerve and at the two time points studied, and were even similar to the polypeptide distribution determined for fast transport.


Afferent and efferent connections of the parabigeminal nucleus in cat revealed by retrograde axonal transport of horseradish peroxidase. Afferent and efferent connections of the parabigeminal nucleus (PBG) of the cat have been demonstrated by means of horseradish peroxidase (HRP) tracing technique. Following HRP injection in the PBG, labelled cells were observed mainly in the deep layers of the ipsilateral superior colliculus (SC). The other labelled structures were the prepositus hypoglossi complex (PH), the ventral nucleus of the lateral geniculate body (LGV), the locus coeruleus, the cuneiform nucleus, the periaqueductal gray and the dorsomedial hypothalamic area. Efferent projections of the PBG were investigated by HRP injection in SC, LGV, PH, hypothalamus and in some acoustic relays, i.e. medial geniculate body and inferior colliculus. Only the PBG-SC projection appeared to be well systematized. The positive labelling of the PBG following injection of LGV and hypothalamus is discussed in terms of the specificity of the injection. The absence of afferent and efferent connections of the PGB with any acoustic relay tends to exclude this nucleus from the auditory system in contrast to previous suggestions. On the basis of the close reciprocal PBG-SC connections a possible role of the PBG within visuomotor tectal function is proposed.


A multiwire proportional chamber study of axoplasmic transport in frog sciatic nerve involving interruption of somatic supply. A multiwire proportional chamber was used to detect axoplasmic transport in isolated spinal-cord-sciatic nerve preparations in which the nerve cell bodies were exposed to L-[35S]methionine. The detector collected radiation from a series of 6-mm segments of sciatic nerve for consecutive periods of 30 min for the duration of experiments lasting 16--23 h. The radioactivity of each segment showed an initial plateau at background level followed by a rise which was approximately linear through time. Plots of the time at which the rise in radioactivity took place against the position of each segment of the nerve yielded transport velocities for the most rapidly moving label; at room temperature (21--23 degrees C) these were typically 150 mm/24 h. Approximately 1.5 h after the addition of 5 mM colchicine to the bathing solution the slope of the radioactivity-time curve decreased for all segments of the nerve; this indicated that the method satisfactorily detected impaired axoplasmic transport. If the sciatic nerve were severed at the proximal end of the preparation at about 9 h after the radioactive front had been established, the slope of the radioactivity-time curve for each nerve segment subsequently changed abruptly. Plots of the time at which the transition occurred against position in the nerve yielded transport velocities which did not differ statistically from those of the radioactive front. This transition time is thought to indicate the time at which the fastest labelled material left each segment of nerve. In some experiments, an additional change in slope occurred, possibly indicating the departure of the slowest material from each segment.


Purification of mouse alpha-foetoprotein by ampholyte displacement chromatography. Mouse alpha-foetoprotein (alphaFP) was isolated from H4 hepatoma tissue using Con A-Sepharose salt gradient ion exchange chromatography and ampholyte displacement chromatography, the latter being a new method for a sharp separation of proteins based on their different isoelectric point. The purity of the alphaFP was demonstrated by (i) the absence of contaminant on sodium dodecyl sulphate polyacrylamide gel electrophetic gels, (ii) Ouchterlony's immunodiffusion against monospecific antimouse alphaFP and the absence of precipitation against a polyvalent antinormal mouse serum, (iii) the production of a monospecific antiserum in a rabbit after injection of the purified antigen, and (iv) immunological unreactivity of the produced antiserum against normal hepatic tissue.


A method for the detection and analysis of growth patterns of microorganisms in soil. A fluorescence-staining technique using the magnesium salt of 8-anilino-1-naphthalene sulfonic acid is described and used to follow the changes in the distribution patterns of microorganisms in soils. A statistical procedure was used to determine the occurrence of significant differences in clumping of bacteria (i.e., production of colonies) in different regions of artificial soil-aggregate systems treated with nutrient solutions and also with a herbicide, Linuron. The response of soil microorganisms to glucose amendment was most marked in the aerobic, outer zone of aggregates. Linuron inhibited colony formation in aggregates treated with the herbicide. The method allows continued observations to be made on the same soil sample at intervals during incubation and os can be used to determine growth rates, inhibitory effects of chemicals, distribution patterns in soils, effects of added nutrients, and other effects where growth in situ is important.


Use of fluorochromes for direct observation of microorganisms associated with wheat roots. The use of several fluorochromes and staining methods was tried out for observation of microorganisms on the surface of wheat roots sampled from soil. Good contrast between fluorescence of microorganisms and fluorescence of root and soil structures was obtained after successive staining of roots with coriphosphine (1:2000, 15 min), congo red (1:10 000, 10 min), and acridine-orange (1:10 000, 2 min).


Human chorionic gonadotropin (HCG) and alpha-fetoprotein (AFP) in sera and tumor cells of patients with testicular seminoma: a prospective study. The HCG and AFP have been quantitated in the sera of 130 patients with the diagnosis of testicular seminoma utilizing a specific double antibody radioimmunoassay. These tumor markers also were localized in tumor cells of some of these patients utilizing an indirect immunoperoxidase technique. 11 of 130 patients had elevated serum levels of HCG. The HCG molecules have been localized in the syncytiotrophoblastic giant cell (STGC) that is occasionally observed in seminomas. None of the patients with pure seminoma had an elevated level of serum AFP. We have concluded that in patients with pure seminoma the level of serum HCG can be elevated (10 of 130 or 7.6%), but we have not observed elevated serum levels of AFP in these patients.


Biochemical and immunological characterization of a reverse transcriptase from human melanoma tissue. An RNA-direct DNA polymerase was purified from human melanoma tissue by successive column chromatography on DEAE-cellulose (DE-23 and DE-52) and phosphocellulose. The purified reverse transcriptase has a mol. wt. of 68,000, a pH optimum of 8.0, a Mn2+ optimum of 0.6 mM, and a KCl optimum of 60 mM. The purified enzyme transcribes (rA)n - (dT)12, (rC)n - (dG)18, (Ome-rC)n - (dG)18 and a 70s RNA from Rauscher leukemia virus (RLV), but failed to transcribe (dA)n - (dT)12. This enzyme has no terminal deoxynucleotidyl transferase activity. Serological studies have shown that the reverse transcriptase from human melanoma tissue is antigenically not related to DNA polymerases from Simian sarcoma virus (SiSV), Avian myeloblastosis virus (AMV), RLV, and human spleen of a patient with myelofibrosis. The purified enzyme showed a close antigenic resemblance to DNA polymerases from baboon endogenous virus (BEV) and rhabdomyosarcoma virus (RD-114), the endogenous virus of the cat.


Isoprinosine: an overview.

Isoprinosine: an overview. Isoprinosine appears to have a relatively low degree of both acute and chronic toxicity in both rodent and nonrodent species. Preclinical studies have demonstrated that isoprinosine can inhibit the growth of both DNA and RNA viruses as well as potentiate cell-mediated immune response both in vitro and in vivo. Clinical studies with rhinovirus, herpesvirus, and influenza virus infections in man demonstrated that isoprinosine treatment reduced clinical symptoms and enhanced certain cell-mediated immune responses compared to placebo-treated controls.


Isoprinosine increases the antitumor action of interferon. The association of relatively low doses of interferon with an immunostimulant (isoprinosine) enhances the antitumor effect of interferon. After combined treatment, the mean survival time, tumor incidence, and final survival rate are significantly increased in mice inoculated with 10(6) Crocker tumor 180/TG cells when compared to mice treated with interferon alone.


Preliminary characterization in mice of the effect of isoprinosine on the immune system. A preliminary characterization of the immunomodulatory activity of isoprinosine in mice was conducted in both normal and tumor-bearing mice. Under the experimental conditions employed, isoprinosine given ip and orally was found to possess some immunomodulatory capacity in various  model systems. The compound appeared to preferentially influence T- but not B-cell responses; it did not apparently influence the functional capacity of macrophages, K cells, and NK cells.


Effects of isoprinosine, levamisole, muramyl dipeptide, and SM1213 on lymphocyte and macrophage function in vitro. Four immunopotentiators, levamisole, isoprinosine, muramyl dipeptide, and SM1213, were analyzed in vitro for their effect on phytohemagglutinin-induced lymphocyte proliferation and lymphokine-induced macrophage proliferation and activation to kill Listeria monocytogenes. Isoprinosine augmented all three functions. Levamisole augmented lymphocyte proliferation and lymphokine-induced macrophage activation. Muramyl dipeptide and SM1213 induced macrophage activation directly. The effects of these agents on these cells at concentrations relevant to in vivo therapy support their immunopharmacologic application to modify cellular immunity.


Electron microscopic study of the orthograde axonal transport of horseradish peroxidase in the supraoptico-neurohypophysial tract of the rat. Horseradish peroxidase (HRP) has been used as a protein tracer in order to visualize the ultrastructural sites of the orthograde transport of protein macromolecules in the hypothalamo-neurohypophysial tract of the rat. After a local injection of HRP within the supraoptic nucleus, the reaction product was observed: (1) mainly in tubules of the smooth endoplasmic reticulum in the more proximal part of the axons, and (2) in granules and microvesicles of the axon terminals. Observations on thick sections clearly showed the existence of a relationship between the smooth endoplasmic reticulum containing HRP and the labeled granules or microvesicles. These data are in good agreement with previous findings showing the existence of direct continuity between tubules of the smooth endoplasmic reticulum and a fraction of the neurosecretory granules and microvesicles. This evidence further reinforces the hypothesis that the latter organelles may possibly originate locally in the axons from the tubules of the smooth endoplasmic reticulum which may therefore be proposed as a possible vehicle for a non-granular intra-axonal transport of neurosecretory material in neurosecretory neurons.


Horizontal cells of the carp retina: Golgi impregnation and Procion-Yellow injection. Application of two methods for the selective staining of neurons, Golgi impregnation and intracellular marking with Procion Yellow, has revealed the anatomical arrangements of the horizontal cells in the carp retina. There are two basic horizontal-cell types, those with axons and these without. The former can be subdivided into three groups on the basis of the pattern of branching of the dendrites. These three structural groups are also functionally distinct, as shown by the electrical recordings made during Procion-Yellow injection. The axons of these three types of cell project into the proximal part of the inner nuclear layer, where they expand to form morphologically indistinguishable terminals. Fine horizontal processes leave the surfaces of these axon terminals. The functional behavior of a terminal resembles that of the associated soma. The horizontal cells lacking axons vary in morphological appearance, but they are similar functionally.


Cyclic AMP in relation to proliferation of the epidermal cell: a new view. Four agents known to increase the level of cellular cAMP by different means (cholera toxin, dibutyryl cAMP, methyl isobutyl xanthine and isoproterenol) increase the growth of colonies of cultured human epidermal cells and of keratinocytes derived from other stratified squamous epithelia. This effect is due to an increase in the overall rate of cell proliferation in the colonies. When added to cultures under hitherto optimum conditions for epidermal cell growth [in the presence of supporting 3T3 cells and epidermal growth factor (EGF)], most of the agents exert an effect of considerable magnitude, the toxin being the most potent. Since the toxin exerts an effect in the absence of supporting 3T3 cells, it must be able to act directly on the keratinocytes. It can also act in the absence of ECF and of medium conditioned by 3T3 cells, although proliferation is greatest when supporting 3T3 cells and EGF are present. The increased proliferation in the presence of the toxin is associated with an increased proportion of small cells known to include the multiplying fraction. The use of toxin makes the cultivation of keratinocytes from epidermis and other stratified squamous epithelia much easier and prolong the culture life of the cells. Whether cell proliferation in the intact epidermis is regulated through agents affecting cAMP (in a direction opposite to that suggested by much of the earlier literature) remains to be elucidated, but the existence of such a mechanism in cultured cells suggests that it may function in the intact epithelium.


Co-regulation of type C RNA virus production and cell differentiation in myeloid leukemic cells. Mouse myeloid leukemic cells which differ in their competence to be induced to differentiate by the normal macrophage- and granulocyte-inducing protein MGI have been used to study the relationship between type C RNA virus production and myeloid cell differentiation. Clones which can be induced by MGI to form Fc and C3 rosettes, to synthesize and secrete lysozyme and to differentiate to mature macrophages and granulocytes (MGI+D+) were induced by MGI to produce higher amounts of type C virus. Clones (MGI+D-) that were less inducible by MGI for Fc and C3 rosettes and lysozyme and were not induced to from mature cells were also less inducible higher virus production. In both types of clones, the increased virus production induced by MGI preceded the induction of rosettes and lysozyme. Clones that were not induced by MGI for rosettes or lysozyme (MGI-D-) showed little or no enhancement of virus production. MGI did not affect virus production in erythroleukemic cells, and erythropoietin did not affect virus production in the myeloid leukemic cells. Dexamethasone, lipopolysaccharide, dimethylsulfoxide and low concentrations of actinomycin D can induce some differentiation-associated properties in some of the clones. With these compounds, there was also a direct relationship between the enhancement of virus production and induction of differentiation-associated properties. Virus released from the three types of clones before or after treatment with MGI or dexamethasone was identified as N-tropic. The enhancement of virus production, as measured by reverse transcriptase activity, was accompanied by an increase in the amount of the viral protein p30, and interferon, which idd not inhibit the induction of differentiation in the myeloid leukemic cells, also did not prevent the increase in the amount of p30. After the early enhancement of virus production associated with the induction of differentiation, a shut-off of virus production occurred in the mature cells induced by MGI in MGI+D+ clones, whereas clones that did not differentiate to mature cells continued to produce virus. The results indicate that enhancement of virus production appears to be an early step in the induction of differentiation. Once induction has occurred, the lack of virus production in the mature cells suggest that a subsequent shut-off of virus production may be required for the completion of differentiation to mature cells. This relationship between cell differentiation and virus production suggests that type C virus has a regulatory role in myeloid cell differentiation.


An avian oncovirus mutant (SE 21Q1b) deficient in genomic RNA: biological and biochemical characterization. We have isolated a nonconditional mutant of PR-RSV-E with unique properties. This virus (SE 21Q1b) is shed from a continuously growing culture of transformed quail cells. 21Q1b virions are unable to transform or replicate in other quail or chicken cells after exogenous infection, despite the fact that the viral particles contain normal envelope glycoproteins, internal structural proteins and RNA-dependent DNA polymerase. The lack of infectivity of 21Q1b virions is a consequence of the failure to package genomic 39S RNA. Instead, these virions contain a mixture of heterogenous-sized polyadenylated cellular RNAs and 4S RNA. Less than 1% of the encapsulated RNA is viral-specific, although in the 21Q1b-producing cells, amounts of 39S, 28S and 21S viral RNAs comparable to those in wild-type virus-infected cells are synthesized and function as mRNAs for the viral proteins. Thus 21Q1b can be considered an RNA packaging mutant. Superinfection of 21Q1b cells with either RAV-1 or PR-A leads to production of about 10% or more of the normal titer of superinfecting virus, but none of the 21Q1b genetic markers are rescued. After superinfection, the 21Q1b cells continue to synthesize 21Q1b particles containing cellular RNAs in the same amounts as before infection. Thus superinfection does not appear to "switch off" the aberrant packaging of cellular RNA, but allows packaging of the superinfecting RNA. One explanation for the phenotype of 21Q1b is that the genome is lacking a signal necessary for efficient genomic RNA packaging (but not for translation) and that the 21Q1b genome encodes a "packaging factor" with an altered specificity so that cellular RNAs are efficiently packaged. 21Q1b virions do contain RNA-dependent DNA polymerase which has normal endogenous synthetic activity. The cDNA product made in vitro from detergent-lysed 21Q1b virions hybridizes equally well to uninfected quail and 21Q1b-producing quail cell RNAs, with kinetics suggesting that the endogenous product consists of transcripts of cellular RNAs present in low amounts in the cells.


[Yolk sac tumor (author's transl)].

[Yolk sac tumor (author's transl)]. An analysis was made from a histopathological and clinical standpoint of 6 cases of yolk sac tumor over a period of 15 years, treated in the first surgical department of the University of Hokkaido. All these cases, with the exception of one retroperitoneal tumor, were diagnosed as embryonic carcinomas before the study. AFP were examined throughout the duration of treatment and their levels reflected changes corresponding to the effects of treatment and recurrence. They were seen in the form of special fluorescence in the tumor cells using the antibody immunofluorescence method. Radiotherapy and chemotherapy may be effective, but the prognosis of these tumors is poor. Amongst the cases mentioned, only the child with a tumor of testicular origin is in good health.


[Isoelectric focusing of the serum vitamin D transport protein (DBP) and the polymorphism of Gc protein. Significance in anthropology]. The authors confirm the analogy between Gc and DBP as a result of analytic studies by isoelectrofocusing and labeling of Gc with 3H vitamin D3. The presence of a relationship between Gc polymorphism and the amount of sunshine in a given region is discussed.


Di-, mono- and nonphytanyl triglycerides in the serum: a sensitive parameter of the phytanic acid accumulation in Refsum's disease. The relative proportions of the diphytanyl, monophytanyl and nonphytanyl triglycerides ("triglyceride pattern"), the phytanic acid content of the triglycerides and the phytanic acid levels in the serum of 3 patients with Refsum's disease (heredopathia atactica polyneuritiformis, phytanic acid storage disease) were estimated by thin-layer chromatography, densitometry and gas chromatography, respectively. The individual triglyceride patterns were clearly dependent on the phytanic acid content of the triglycerides: the more phytanic acid in the triglycerides, the higher the percentage of the diphytanyl and the lower the percentage of the nonphytanyl triglycerides. The monophytanyl triglycerides were also related to the phytanic acid content of the triglycerides, although in a complex manner. The triglyceride pattern can be taken as a parameter of the phytanic acid accumulation in Refsum's disease to be at least as sensitive as the serum phytanic acid level in the biochemical evaluation of the efficiency of the dietary treatment.


Cri-du-chat syndrome in a child with a 5/15 translocation and interstitial centromeric heterochromatin. A child is described with most of the typical clinical features of the cri-du-chat syndrome. G- and C-banding studies revealed the karyotype 45,XX, -5, -15, +tdic (5;15) with the loss of short arm material from chromosome 5. Centromeric heterochromatin of the translocated No. 15 is still present in the translocation chromosome. However, no silver precipitation after AgNO3-staining was observed on the translocation chromosome, thus indicating a loss or genetic inactivation of the NOR-region of the translocated No. 15. These cytogenetic results and their possible relationship to the cri-du-chat phenotype are discussed.


Prenatal diagnosis of the Meckel syndrome. Prenatal diagnosis of the Meckel syndrome was made at 20 weeks of gestation from the findings of a biparietal diameter smaller than expected for gestational age, a grossly raised amniotic fluid alphafetoprotein level and a rapid growth of foetal macrophages after 20 hours culture. Termination at 23 weeks of gestation resulted in a male foetus with an occipital encephalocele, microcephaly, polydactyly, and bilateral polycystic kidneys. This case report emphasies the importance for genetic counselling of delineating the Meckel syndrome from the multifactorial cases of neural tube defects, and also illustrates, at least in some cases, that the syndrome can be diagnosed in utero.


Hand metastasis from melanoma: a case study. Metastases to the hand are rare. Most frequently they result from carcinomas of the lung, breast or kidney. This is a case report of a rare peripheral metastatic lesion in a 24-year-old man with melanoma. There are no previously reported hand metastases from melanoma. Because of the extremely poor prognosis for patients with osseous metastases from melanoma, treatment should be palliative.


Significance of the apexcardiogram in non-invasive assessment of postextrasystolic potentiation. The study  revealed the usefulness of the apexcardiogram in the assessment of the contractile state of left ventricle in cases of ventricular extrasystolia. The pre-ejection period/left ventricular ejection time (PEP/LVET) ratio and isovolumic contraction period (IVCP) are good parameters for demonstrating the postextrasystolic potentiation, the lack of which occurs in severe heart failure. PEP/LVET and IVCP are R-R-interval dependent in parasystolia. In the establishment of the postextrasystolic potentiation, the authors contribute an important role to the ventricular filling during the compensatory pause.


Effects of various drugs on the osmotic lysis of rat mast cells. 1. Rat mast cells were exposed to low osmotic pressures to produce a 'non-specific' disruption of the mast cell, with release of histamine along with other intracellular contents. 2. The effect of non-steroidal anti-inflammaotry drugs and various other drugs upon osmotically induced histamine release was examined. 3. Representative non-steroidal anti-inflammatory drugs indomethacin, phenylbutazone and flufenamic acid, the acidic compounds ethacrynic acid, iopanoic acid and probenecid, and the local anaesthetic lignocaine, all caused a dose-dependent facilitation of osmotically induced histamine release compared to controls. All drugs were active at 0.1 mmol/l. 4. The previously observed inhibition of compound 48/80 and the antigen-induced histamine release from rat mast cells by similar concentrations of the drugs used in the present study are unlikely to be due to mast cell plasma membrane stabilization.


Techniques and therapies for primary and metastatic liver cancer. In summary, of the 460 patients of primary carcinoma of the liver admitted to the University Surgical Unit at the Queen Mary Hospital over a period of 12 years, more than 40% could not be treated, and only 91 of the patients were candidates for curative resection. The cure rate is very small; a 1- to 2-year survival was obtained in 46% of 15 resections. From 1964 to 1969, out of 22 patients with resections, 3 are still alive more than 5 year after the operation. Lin30 reported a 19.1% 5-year survival. When the hepatoma has ruptured and bleeding takes place, surgical treatment is obligatory to control the hemorrhage. Ninety-eight patients underwent a clinical trial of 5 categories: hepatic dearterialization, hepatic arterial cannulation and infusion of 5-FU, hepatic arterial ligation and portal venous infusion of 5-FU, radiotherapy and no treatment. The results show that the advantage of each form of treatment when compared with no treatment is marginal. Thus a gloomy picture of primary hepatoma is held. Since the operative mortality of hepatic resection for a solitary secondary carcinoma of the liver is negligible, it should be done in each instance because a long-term survival may be possible. This is especially true with primary carcinoma of the colon.


Treatment of Kaposi's sarcoma with bleomycin. Two cases of Kaposi's sarcome that responded to bleomycin therapy are reported herein. The first case, resistant to various treatments, responded remarkably well after 240 mg of bleomycin was administered over a thirteen month period. The second case had rapidly progressive untreated Kaposi's sarcoma of six months's duration; a prompt response was obtained to an initial course of 105 mg of bleomycin.


Anatomical and functional aspects of the septo-hippocampal projections. The origins, distribution, and cellular targets of the septo-hippocampal projections are reviewed. It appears that the distribution of acetylcholinesterase-positive neurons in the medial septum and diagonal bands and those cells labelled after injections of horseradish peroxidase into the hippocampus coincide; however, the possibility of a non-acetylcholinesterase septal projection remains. Good agreement is found between the distribution of hippocampal acetylcholinesterase and the patterning of silver grains after injection of [3H]leucine into the medial septum. A major target of septal efferents to the hippocampus is the interneuron population; the possibility of septal mediation of intrahippocampal circuitry via this anatomical arrangement is discussed.


[Histamine levels in blood of patients with allergic bronchial asthma (author's transl)]. The investigation of 10 patients with allergic asthma showed histamine levels of 79.8 +/- 50.0 ng/ml in blood during symptom-free intervals. They were not significantly different from those in 12 patients with chronic obstructive bronchitis (54.0 +/- 25.9 ng/ml) and in 13 healthy probands (56.9 +/- 21.5 ng/ml). It could be shown that allergen-induced bronchial obstruction in patients with allergic asthma had no demonstrable influence on blood histamine levels. The absence of increased blood histamine levels in allergic bronchial obstruction supports the assumption that allergen-induced liberation of histamine and its biological effects are mainly limited to the bronchial system. Local metabolisation of the liberated mediator can be supposed to a large extent.


[Percutaneous drainage of the biliary tract by means of a fine needle (author's transl)]. Temporary pre-operative and post-operative as well as permanent palliative percutaneous biliary-tract drainage was performed in 12 patients with biliary tract obstruction; in three the drainage was internal and in nine external. Main indication of the method is permanent palliative decompression in cases of inoperable malignant tumour with obstructive jaundice. Pre-operatively, percutaneous biliary tract drainage serves as a temporary measure in order to perform an operation after decompression of the biliary tract and improving the patient's general condition. The only serious complication was biliary peritonitis after a drainage catheter had slipped out.


[The effect of cytostatic therapy with bleomycin and methotrexate on cellular immune reactivity]. Considerable importance for the elimination of malignant cells has been attributed to the cellular immune system. Cytostatic drugs however are known to be immunosuppressive. The transformation response of peripheral lymphocytes to bleomycin and methotrexate was determined before and after treatment in patients with squamous cell carcinoma of the head and neck region. The transformation response was interpreted as an indication of the potential for the nonspecific cellular immune reaction. The cellular reaction was significantly suppressed during therapy; this suppression was then followed by an excessive reaction in some patients. The immunosuppressive effect of bleomycin was substantiated in in vitro studies of isolated lymphocytes from healthy doners. Low concentrations of methotrexate however produce excessive reactions while higher concentrations tend to have an immunosuppressive effect.


Recognition of idiotypes in lymphocyte interactions. I. Idiotypic selectivity in the cooperation between T and B lymphocytes. Idiotypic restrictions are demonstrated in vitro for the cooperation between T and B lymphocytes with specificity for Group A streptococcal carbohydrate. T helper cells which have been primed in vivo with anti-idiotypic antibodies to the A5A idiotype and which are therefore essentially A5A idiotype-positive, cooperate only with A5A idiotype-positive B cells, even when mixtures of A5A idiotype-positive and A5A idiotype-negative B cells are present. Essentially A5A idiotype-negative T helper cells that have been primed in vivo with Group A streptococcal vaccine after in vivo suppression with anti-A5A idiotypic antibody are unable to cooperate with B cells which have been primed with anti-A5A idiotype antibody and which are therefore essentially A5A idiotype-positive. Mixtures of A5A idiotype-negative and A5A idiotype-positive T cells cooperate with both A5A idiotype-negative and A5A idiotype-positive B cells. Idiotypic restrictions could not be demonstrated for T and B cells recognizing carrier and hapten determinants, respectively, in experiments in which the cooperation of genetically VH-identical T and B cells was compared to the cooperation of genetically VH-different T and B cells. The data are discussed with respect to various models for the communication between T and B cells. It is proposed that for successful T-B cooperation, ordinarily two types of T helper cells are required, one recognizing the antigen and the other recognizing the idiotype of the B cell.


Some chronic lymphocytic leukemia cells bearing surface immunoglobulins share determinants with T cells. One set of antigens is common to some chronic lymphocytic leukemia (CLL) cells bearing surface immunoglobulins (Ig) and normal T cells. Proliferating cells from thirty-eight patients were studied with four antisera recognizing normal human T but not B cells. These antisera were raised in rabbits against (a) Sezary cells, (b) blood lymphocytes from a patient with sex-linked agammaglobulinemia, (c) T lymphoblasts and (d) thymus cells. In four CLL cases, the cells expressed the receptor for sheep erythrocytes and lacked surface Ig. Cells from thirty-four CLL cases bore monoclonal surface Ig and did not bind sheep erythrocytes. Twelve out of these thirty-four cases of CLL had cells which were lysed by one or, more frequently, by the four anti-human T cell xenoantisera. By absorption experiments, one set of at least three antigens common to the cells of some of these CLL and T cells was defined. Depending on the patient, the cells can either carry one, some or all of the antigens of this set. However, it was also demonstrated by absorption that these cells lacked antigens particular to the T cell lineage, while the cells from T CLL cases carried both sets of antigens.


H-2-linked murine cytotoxic T cell responses specific for sendai virus-infected cells. CBA (H-2k) mouse-derived lymphochoriomeningitis virus and herpes simplex virus-specific cytotoxic T lymphocytes lyse virus-infected target cells compatible on either the H-2k or H-2D region. In contrast, CBA, C3H and AKR (H-2k) mouse-derived sendai virus-specific cytotoxic T lymphocytes (CTL) fail to lyse H-2D-compatible virus-infected cells. A similar lack of H-2D region-associated lytic activity was found with C57BL/6 and C57BL/10 (H-2b) mice as well as with the recombinants B10.A (2R) [Kb-Db] and B10.A (4R) [Kk-Db]. On the other hand, BALB/c (H-2d) mice and A/J (H-2a) mice do generate H-2Dd-associated sendai virus-specific CTL. These results are in contrast to those obtained with (CBA X BALB/c)F1 and B10.HTT [Ks-Dd] mice, which failed to mount Dd region-associated CTL responses. It is concluded that D region-associated sendai virus-specific CTL responsiveness varies with the H-2 genotype of the responder cells.


Interaction between barium, strontium and calcium in histamine release by compound 48/80. Pre-incubating rat peritoneal and pleural mast cells for 30 min at 2 degrees C in Ca2+-free Tyrode containing Ba2+ (30 mM) markedly reduced histamine release by compound 48/80 (0.5 microgram/ml). Evoked secretion was not significantly altered by pre-incubation with Sr2+ (30 mM) at this temperature whilst Mg2+ (30 mM) and La3+ (0.1 mM) slightly inhibited and potentiated release respectively. The effect of Ba2+ was concentration related and progressively reversed by Ca2+ (1--10 mM). Pre-incubation at 37 degrees C in Ca2+-free medium alone substantially reduced histamine release by compound 48/80. This reduction was unaltered by either Mg2+ (30 mM) or La3+ (0.1 mM), prevented by either Sr2+ (30 mM) or Ca2+ (1 mM) and augmented by Ba2+ (30 mM) which also markedly increased spontaneous histamine release during pre-incubation at 37 degrees C. Results suggest that Ba2+ and Sr2+ interact with cell-fixed calcium to modulate histamine release by compound 48/80.


Projections of the precentral motor cortex and other cortical areas of the frontal lobe to the subthalamic nucleus in the monkey. Corticofugal projections were examined by means of the autoradiographic tracer method in 21 macaca fascicularis. The labeled material was injected into the main body representation areas of the precentral motor cortex and into various regions of Brodmann's areas 6, 8 and 9 of the frontal lobe. The ipsilateral subthalamic nucleus receives a moderately strong and somato-topically organized projection from Woolsey's precentral motor cortex. This projection is mainly restricted to the lateral moiety. The remaining nucleus is occupied by less intensive projections from premotor and prefrontal areas. It is concluded that the subthalamic nucleus is a convergence site of pallidal and corticomotor and frontal projections. Cortical afferents may exert an influence on the pallido-subthalamic-pallidal inhibitory feedback loop.


Nigrothalamic pathway in the cat demonstrated by autoradiography and electron microscopy. Light microscopic autoradiography and electron microscopy were used to trace the nigrothalamic projections and to study the sites of termination of this pathway in the cat. Injections of tritiated amino acids or electrolytic lesions were placed in the substantia nigra pars reticularis (SNr). An accumulation of radioactivity was found in the ventral medial nucleus and in the ventromedial part of the ventral anterior nucleus. At the ultrastructural level degenerating medium size synaptic boutons and medium size myelinated fibers were observed in these nuclei. The boutons contained clear pleomorphic veiscles and formed symmetrical type synaptic contacts with regular type dendrites and vesicel-containing dendrites. The present findings indicate that the ventral medial nucleus is the principal site of termination of nigrothalamic projections in the cat.


Identification of the projection from the visual cortex to the claustrum by anterograde axonal transport in the cat. Visual cortex, including areas 17, 18, and sometimes 19, was injected with tritiated leucine. Terminal labelling could be detected by autoradiography in the dorsocaudal part of the ipsilateral claustrum in all cases.


Projections of precentral and premotor cortex to the red nucleus and other midbrain areas in Macaca fascicularis. Midbrain projections from the precentral and premotor cortex were studied autoradiographyically in seven macaques. Earlier findings of Kuypers and Lawrence (1967) based on silver impregnated axonal degeneration were largely confirmed and partly extended. Laminar projections into the parvocellular red nucleus were found bilaterally from the somatotopic representation areas of the precentral cortex and three main divisions of premotor cortex (ventral, dorsal, and medial). The heaviest contralateral projection was recorded from the medial hemispheric aspects of area 6 (partly identical with the supplementary motor cortex). Homolateral projections were found from the limb areas of the precentral motor cortex to the magnocellular portion of the red nucleus and direct evidence for the overlap between corticorubral projections and corresponding areas of origin of the rubrospinal tract was obtained. Other projections concerned N. Darkschewitsch, N. Edinger-Westphal, suprarubral and dorsal mesencephalic reticular formation and adjacent lateral periaqueductal grey, substantia nigra, pretectal region, superior colliculus and N. tegmenti pedunculo-pontinus.


Effect of growth-promoting alpha-globulin on the incorporation of exogenous DNA into L cells. II. Fate of exogenous 3H-DNA after incorporation into recipient cells. GPAG stimulates the uptake of exogenous 3H-DNA by L cells and facilitates its retention in host cell nuclei during 48-h postcultivation; the nuclei contain 1.5 times more radioactivity than GPAG-free controls. Owing to the well-known properties of GPAG, the formation of the 3H-DNA-GPAG complex may affect the incorporation of exogenous 3-H-DNA and its intracellular transport and retention in the host cell nuclei.


Antral gastrin cell proliferation after vagotomy in rats. The total number of antral gastrin cells, the antral surface area and the serum gastrin concentration were determined in two groups of rats. The animals had been submitted at random either to vagotomy and pyloroplasty or to pyloroplasty alone 3 weeks before. The indirect immunoperoxidase reaction and a direct quantitative histological method were used to count the gastrin cells. Radioimmunoassay was used to estimate the serum gastrin concentration. The total number of gastrin cells in the stomach was 52% (p less than 0.01) more elevated in the vagotomized animals. Both antral mucosal surface and calculated concentration of gastrin cells per square millimeter of mucosa were significantly higher (0.01 less than p less than 0.05) in this group of animals. The serum gastrin values were significantly (p less than 0.01) more elevated after vagotomy. These observations indicate that vagotomy may induce an antral gastrin cell hyperplasia which could explain in part the hypergastrinemia observed after this surgical procedure.


Intragenic suppression at the b2 locus in Ascobolus immersus. I. Identification of three distinct groups of suppression. A reversion study of EMS- or ICR170-induced ascospore color mutants in Ascobolus immersus is reported. Twenty-three new intragenic suppressors were isolated within the b2 locus. These are localized within three distinct groups on a genetic fine-structure map containing 21 identifiable sites. The pattern of reversion is discussed according to the known specificity of EMS and ICR170 in Ascobolus immersus.


[Lymphocyte transformation test--basis and clinical use]. The principle of the lymphocyte transformation test is represented and the present knowledge of the partially selective effect of various unspecific mitogenes on different lymphocyte populations is referred to. Problems of methodology are briefly dealt with and the possibilities of applying the lymphocyte transformation test is referred to in detail. Own experiences are reported on the basis of clinical examples in patients affected with malign lymphoma, infectious mononucleosis and polymyositis.


Specificities of heterologous antisera against human leukaemia cells. 1. Reactions against leukaemia cells. Rabbit or goat antisera directed to ALL, CLL, AML and CML cells were investigated in cytotoxicity tests with different leukaemia and normal cells as targets. After absorptions with erythrocytes and spleen cells from allogeneic donors the antisera killed only leukaemia cells. There was no reaction with remission leukocytes or blood leukocytes from normal donors. Anti-ALL-Sera reacted in 35 out of 49 tests with ALL cells from 13 patients. Apparently the ALL antisera which were directed to the T cell subtype of ALL preferentially affected ALL cells of this subtype. Cross reactions with cells from CLL, AML and CML were not found. Anti-CLL-sera reacted in 10 out of 12 tests with CLL cells from 4 donors, and in 4 out of 20 tests with ALL cells from 7 donors and also with the cells of a CML patient. AML cells from two patients were not killed. Antisera against AML and CML showed extensive cross reactions with cells of myelocytic and lymphocytic leukaemias. Absorption tests demonstrated the presence of two antibody specificities in AML antisera, one of which being directed to a common antigen of AML and ALL cells and another against an antigen of myelocytic leukaemia cells.


Specificities of heterologous antisera against human leukaemia cells. 2. Reactions against fetal liver cells and absorption studies with fetal tissue. Rabbit or goat antisera directed to ALL and AML cells were investigated in cytotoxicity tests with fetal liver cells as targets. After absorption with erythrocytes and spleen cells from allogenic donors the antisera killed fetal liver cells. There was no reaction with remission leukocytes or blood leukocytes from normal donors. Treatment with fetal tissue removed the activity of the AML and ALL antisera against ALL cells but not of the AML antisera against AML cells. This indicates the existence of at least two antigens on the surface of AML cells, one antigen is common with ALL cells and of fetal origin and another one seems to be characteristic of AML cells and not of fetal origin. Because treatment with fetal tissue removed all activity of the ALL antisera it can be assumed that leukaemia-associated antigens on ALL cells are of fetal origin.


Is granulopoietic activity (GA) in vivo comparable to colony stimulating factor (CSF)? Human sera after column chromatography on Sephadex G-200 were tested for Granulopoietic Activity (GA) in vivo and Colony Stimulating Factor (CSF) in vitro. We have found that GA after chromatography is distributed in three peaks but CSF only in one peak with high activity, and evident activity in volume 135 ml. This activities are in good agreement with two peaks of GA. We assume that GA and CSF after this chromatographic procedure is free from lipoproteins inhibitors. The obtained results are briefly discussed.


Activity of acid and alkaline phosphatase in neutrophils of rats exposed to benzene and treated with selenium. Cytochemical studies on activity of acid phosphatase (AcP) and alkaline phosphatase (AP) in peripheral blood neutrophils of rats chronically exposed to benzene vapours (1,200 mg/cm3) demonstrated that the exposure results in the increase of AcP and the decrease of the AP activity. The changes noted correlate with the time of exposure and are observed even after 5 months after exposure. The administration of sodium selenate in a dose of 1.0 microgram/kg before exposure prevented the above enzymatic alterations caused by benzene. In contrast, the administration of selenium in a dose of 5.0 microgram/kg only prevented the change of AP activity and caused reactive neutrophilic leukocytosis.


The depressing effect of tumour on bactericidal capacity of plasma and leukocytes. Leukocyte--plasma suspensions and plasma from tumour patients have been tested in vitro against staphylococcus aureus for bactericidal capacity. Blood was collected during surgical intervention from peripheral vein and from the nearest point of vein to tumour. Second collection of peripheral blood was done 2--3 weeks later after surgery. Decrease of bactericidal activity of plasma has been found in blood collected from vein near the tumour. We can suggest that tumours release some factors disturbing the bactericidal capacity or that the tumour absorbs some factors of plasma or changes the functions of granulocytes.


The transient loss of HLA antigens on lymphocytes in patients following administration of penicillin and tetracyclin. Ten patients with inflammatory affections of urinary, respiratory and bile tract, as well as otitis patients were administered Penicillin. Six or seven days after the start of the treatment, four of them showed weakening or loss of minimally two earlier determined HLA antigens. The loss of HLA antigen was likewise demonstrated in a patient treated with Tetracycline. The HLA antigens were reliably demonstrated again six to seven days after the end of the treatment. The results suggest a possible false typing of HLA antigens in patients who had been administered Penicillin or Tetracycline. In accordance with Ben-David et al. [1] transient loss of two HLA antigens with concomitant development of lymphocyte polyreactivity was found in one of four patients treated with Chloramphenicol.


Enhancement of the specific cytotoxic reactivity of HLA antigens on lymphocytes stored at +4 degrees C for 24 hours. The lymphocytes of some patients (especially with malignancies) as well of healthy subjects, which had been isolated on the day of blood withdrawal from the heparinized blood, show negative or slightly positive reaction with some corresponding HLA sera; after 24 h of storing in the refrigerator at +4 degrees C the lymphocytes tend to restore the cytotoxic reactivity of HLA antigens, and thus, a strongly positive reaction can be recorded. The cause of this phenomenon may be ascribed to the anticomplementary effect of the lymphocyte suspension in barbital buffer.


[Modifications of lymphocyte HL-A antigens as a consequence of therapy in patients with manic-depressive psychosis]. In 54 (= 46.96%) of 115 patients with maniacal-depressive psychosis a HLA modification could be identified. This modification turned out to be temporary and from a serological point of view it revealed a different character. In 29 cases a loss of HLA antigens could be observed, in 3 cases there was a decrease, in 14 cases a combination of both changes, twice a polyreactivity was observed and 6 times a change of the antigen HLA-A 2 in A 28 could be determined. These serological modifications appeared after therapy with lithium as well as with various antidepressive and neuroleptic medicaments. The connection between therapy and development of HLA modification could be ensured statistically. The modifications of HLA antigens A 10 and B 7 developed after administering neuroleptic medicaments, those of HLA antigens A 9, A 11, B 12, and B 13 after therapy with antidepressive medicaments. HLA antigens B 27 and B 40 showed a relative resistance towards therapy. The significance of these findings for the possibility of mistakes in HLA typing and from the standpoint of therapy efficiency in connection with the patient's HLA phaenotype is discussed.


[Effect of vitamin E on the mechanical fragility and the Mg++-ATPase of rabbit erythrocytes]. We have established that Mg++ activated ATPase in red cells of rabbits is not changed by vitamin E injection, even if the mechanical resistance of these red cells is increased. Enzyme activity, however, cannot be correlated with this increased mechanical resistance. Probably, the effect of vitamin E on membrane lipids may be supposed to exist, though its effect on membrane proteins cannot be fully excluded.


Evidence against the presence of "inhibitors" in coumarin treated patients. Coumarin drugs or vitamin K absence cause a decrease of factor II, VII, IX and X activities and the appearance of pre-factors into the circulation. Such pre-factors have been postulated to inhibit thrombin conversion. The current of research on the alleged activity of such "inhibitors" is taken into consideration. Thrombotest discrepancy, mixing experiments etc.) speak against the inhibitor theory. So far the only sure demonstration of the presence of coumarin induced pre-factors has been obtained by immunological means. However, this does not say anything about their biological activity. These pre-factors could well be "inert" as far as clotting is concerned. Until new, unequivocal data on the subject will be available, any method or technique claiming to be able to detect or monitor the "inhibitory" effect should be accepted with extreme caution. Too many unjustified views have been put forward in recent years.


Immunofluorescent evaluation of platelet alpha-1-antitrypsin and alpha-2-macroglobulin. Rabbit raised anti-alpha-1-antitrypsin or anti alpha-2-macroglobulin antisera at dilution of less than 1:80 yielded non-specific staining on human platelets by indirect immunofluorescent technique. A similar pattern was in fact obtained by using normal rabbit sera at the same dilution and was due to the presence of smooth muscle autoantibodies. This indicates that human platelets do not contain significant quantities of these antigens. In agreement with the above, only microamounts of alpha-1-antitrypsin and alpha-2-macroglobulin were found to be present in human platelets by means of the electroimmunoassay.


Protein cross-match test for the diagnosis and prevention of reactions after the transfusion of blood and cryoprecipitate. The examination by means of counterimmunoelectrophoresis for protein incompatibility of the serum of 62 patients who developed an allergic or pyretic post-transfusion reaction revealed incompatibility in 23 cases (37.2%). It was due to the presence of antibodies in 8 recipients (34.8%) and in 15 blood donors (65.2%). The incidence was significantly higher than in a control group of transfused patients who did not develop a reaction. In this group protein incompatibility was found in only 14.7%, 80% of which was due to antibodies in the recipient. In 13 (56.5%) of the patients with reactions agglutinating, cytotoxic or complement fixing antibodies against cellular antigens or IgG were found in addition to protein incompatibility. In 10 cases (43.5%) protein incompatibility was the only explanation for the clinical symptoms. When, in the treatment of multiply transfused haemophiliacs who regularly developed adverse reactions, donors for the preparation of cryoprecipitate were selected by means of the described technique, the almost obligatory reactions were prevented.


[Screening of blood donors for the prevention of post-transfusion hepatitis by means of gamma-glutamyl transpeptidase determination]. The authors describe the application for determining gamma-glutamyl-transpeptidase (GGTP) activity in donors in order to prevent posttransfusion hepatitis. They particularly refer to the high susceptibility of this method for detecting latent and chronic liver diseases virtually unable to be recognized by other methods. Regular GGTP screening having been introduced in the past 4 years, a rapid lowering of the occurrence of posttransfusion hepatitis could be observed. The authors recommend this method of examination as a valuable aid to the prophylaxis of hepatitis transferred by blood preparations.


Infant botulism: new guise for an old disease. While classic botulism is usually due to preformed toxin in improperly prepared food, the newly recognized infant syndrome occurs when spores or vegetative cells germinate in the gut of the very young and elaborate toxin there. The resulting illness may range from inapparent to fulminant, sometimes causing respiratory arrest and death. Infant botulism may also account for at least some cases of sudden infant death syndrome.


Sister chromatid exchange in lymphocytes from patients with malignant lymphoma. Sister chromatid exchange (SCE) frequencies were studied in differentially stained lymphocytes from 47 patients with malignant lymphoma. Thirteen patients were untreated when studied. The mean SCE frequency [+/- standard error (SE)] for these patients was 12.7 +/- 0.9 per mitosis. The mean score for 40 controls was 6.1 +/- 0.3. SCE mean scores were significantly higher in the untreated patients than in the controls (P less than 0.001). Seven patients were treated with radiotherapy alone. They demonstrated a mean SCE frequency (8.8 +/- 0.8) significantly lower (P less than 0.01) than that found in untreated patients. Eleven patients received cyclophosphamide within 4 weeks prior to study. They demonstrated a mean SCE frequency (14.3 +/- 1.3) significantly higher (P less than 0.05) than that found in patients who had received regimens that did not contain cyclophosphamide in the prior 4 weeks (11.1 +/- 1.3) or who had been off drugs for at least 8 weeks (10.1 +/- 0.8). Our data suggest that untreated patients with malignant lymphoma have elevated SCE frequencies, which may be further increased by certain chemotherapeutic agents.


A new immunochemical method for the quantitative measurement of specific gene products in man-rodent somatic cell hybrids. An immunochemical method has been developed for the quantitative determination of species-specific gene products, for instance alpha-galactosidase and N-acetyl-alpha-galactosaminidase, in man-rodent hybrid cells and in the parental cell lines. Antisera raised against the purified enzymes are covalently coupled to Sepharose 4B. The gene products are specifically removed from a cell lysate by incubating with the appropriate Sepharose-coupled antiserum. After centrifugation followed by washing of the precipitated Sepharose, the enzymic activity can be quantitatively measured on the Sepharose beads. With this technique it has been demonstrated that the ability of human N-acetyl-alpha-galactosaminidase (also known as alpha-galactosidase B) to hydrolyze alpha-galactosidic linkages is lost when the enzyme is expressed in man-Chinese hamster hybrid cells.


Differential staining in the secondary constriction regions of human chromosomes A1, C9, and E16 by a heat-hypotonic treatment technique. The secondary constriction (SC) regions of human chromosomes A1, C9, and E16 were preferentially stained with Giemsa when the mitotic cells in a hypotonic solution were heated in a water bath before fixation. The staining intensity of these regions increased from E16 through A1 to C9. The SC region of C9 was stained more intensely as the treatment temperature was raised from 45 degrees C to 60 degrees C, while the staining intensity of the SC region of E16 reached a peak at 50 degrees C and then decreased gradually with increasing temperature. The SC region of A1 showed stable differential staining between 50 degrees C and 60 degrees C. These observations suggest a heterogeneous nature of the SC regions of human chromosomes, probably reflecting a difference in the chromosomal proteins or the DNA-protein interaction in these regions.


Anencephaly in trisomy 18 associated with elevated alpha-1-fetoprotein in amniotic fluid. Elevated levels of alpha-1-fetoprotein (AFP) were found in the amniotic fluid of a 36-year-old woman in the 15th week of gestation. Because of this and the results of repeated ultrasonography, abortion was induced. An anencephalic fetus with trisomy 18 was delivered. The possible correlation among neural-tube defects, chromosomal abnormalities, and level of AFP is discussed.


The association between undifferentiated nasopharyngeal carcinoma and Epstein-Barr virus shown by correlated nucleic acid hybridization and histopathological studies. All of 51 undifferentiated carcinomas of the nasopharynx were EBV-DNA-positive and, in the few tests performed, EBNA-positive as well. Most of these patients also had high anti-EBV [VCA and EA (D)] antibody titres. Of four patients that had somewhat differentiated NPCs, two were EBV-DNA-positive with a corresponding serological picture, whereas two were negative with an EBV serology comparable to that of healthy donors. Of seven lymphomas localized in the nasopharynx, one was EBV-DNA-positive and corresponded histologically to Burkitt's lymphoma, whereas six others were EBV-DNA-negative. Fourteen head and neck carcinomas outside the nasopharynx were all EBV-DNA-negative. These results confirm the consistent and unique association of EBV DNA with undifferentiated carcinomas of the nasopharynx.


Diagnosis and differential diagnosis of lymphoepithelial carcinoma in lymph nodes: histological, cytological and electron-microscopic findings. (1) Lymphoepithelial carcinoma, which occurs in the nasopharynx and palatine tonsils, is a special variant of squamous-cell carcinoma with a non-neoplastic lymphocytic component. (2) The morphology of lymphoepithelial carcinoma is very characteristic if not specific. Therefore, whenever lymphoepithelial carcinoma is diagnosed in a cervical lymph node, the clinician must try to find the primary tumour in the nasopharynx or tonsils, by blind biopsy if necessary. (3) The 'inflammatory' component of lymphoepithelial carcinoma is often very conspicuous. Tuberculoid lesions, with or without caseation necrosis, and marked eosinophilia and plasmacytosis are highly characteristic and help to confirm the diagnosis. (4) The 'inflammatory' component, including the lymphocytes among and around the tumour cells, is not fully understood; but it would appear that T lymphocytes are responsible for all, or most, of the cellular reactions against the tumour.


European Organization for Research on Treatment of Cancer (E.O.R.T.C.). Controlled trials of chemotherapy as an adjuvant or palliative treatment of nasopharyngeal carcinoma. The E.O.R.T.C. Head and Neck Cooperative Group has designed a relatively complex international programme for a multidisciplinary approach to the treatment of NPC. The aim of the programme is to evaluate the usefulness of chemotherapy in preventing metastases and recurrences and, at the same time, to test other chemotherapeutic regimens and to compare them subsequently  with the first one. Two contemporaneous controlled trials are foreseen: (1) long-term chemotherapy after radiotherapy for T3-4 No and T1-4 N1-3 MO cases (NPC 1) and (2) aggressive chemotherapy for MI cases and for patients who develop local or regional recurrences during the first trial (NPC 2). A search of the literature and a pilot study carried out at the Istituto Nazionale dei Tumori in Milan on advanced cases of NPC showed the following percentages of regression after various kinds of chemotherapy: cyclophosphamide, 38%; adriamycin, 39%; bleomycin, 28%; and methtrexate, 17%. On this basis, the following regimens were chosen: NPC 1 trial: cyclophosphamide (1.2 g/m2 every three weeks for 10 cycles) and a control group receiving no chemotherapy NPC 2 trial: adriamycin alone versus a combination of vinblastine + bleomycin + adriamycin. The programme was begun a short time ago, and no results are yet available.


Binding of aggregated human beta2-microglobulin to surface protein structure in group A, C, and G streptococci. A novel mammalian-microbial "short circuit" has been demonstrated between aggregated human beta2-microglobulin and group A, C, and G streptococci. Bacteria belonging to nine gram-positive and three gram-negative species were tested for binding of radiolabeled beta2-microglobulin. All 10 individual strains of group A streptococci showed a high degree of reactivity with aggregated human beta2-microglobulin. Among 27 group C and 28 group  G streptococci, 9 and 6 strains, respectively, were highly reactive, whereas the remaining strains showed a lower, but definite level of beta2-microglobulin binding. Of 11 group B streptococci, 4 were slightly positive. All strains among the other eight species were completely negative. Simultaneous testing of A, C, and G streptococci for immunoglobulin binding showed a lack of correlation between type II and III Fc reactivity and beta2-microglobulin binding. There was no inhibition of uptake of aggregated beta2-microglobulin to reactive strains when excess amounts of human immunoglobulin were added. The beta2-microglobulin-binding surface structure was found to be markedly sensitive to trypsin digestion. The relative trypsin resistance of the immunoglobulin-binding protein in the digestion experiments further demonstrated the dissociation between these two reactivities.


Evidence that L-rhamnose is the antigenic determinant of hyporesponsiveness of BALB/c mice to Klebsiella pneumoniae type 47. The genetic polymorphism in early, immunoglobulin M, responsiveness to the Klebsiella pneumoniae type 47 polysaccharide (K47-PS) is determinant specific, and L-rhamnose is probably the determinant against which differential responsiveness is expressed. This is inferred from tests of the response to the cross-reacting Streptococcus pneumoniae type 23 and non-cross-reacting pneumococcal polysaccharides of known chemical construction. B10.2/Sn new-line mice are high responders and BALB/c mice are low responders to K47-PS. Upon immunization with killed S. pneumoniae type 23, these strains produce cross-reacting antibody to K47-PS. The cross-reacting antibody responses of these strains are congruent with their responses to K47-PS.


External layers of Rickettsia prowazekii and Rickettsia rickettsii: occurrence of a slime layer. Using a simple specific-antibody stabilization procedure on organisms gently liberated from their host cells, we have demonstrated by electron microscopy that Rickettsia prowazekii and Rickettsia rickettsii possess a coat of variable thickness, external to the outer leaflet of the cell wall and the structure designated by others as a "microcapsule," which corresponds most closely to the slime layer of certain other bacteria. Reactions in the methenamine silver and ruthenium red staining procedures and the failure to be visualized by standard procedures suggest that the slime layer is largely polysaccharide in nature. It is postulated that this slime layer accounts in large part for the large, electron-lucent, halo-like zone which is found by electron microscopy to surround organisms of the typhus and spotted fever groups in the cytoplasm of their host cells, that it may be the locus of some major group-specific antigens, and that it may function as an antiphagocytic mechanism, as an aid for attachment of rickettsiae to potential host cells, or both. Moreover, because the attenuated E strain of R. prowazekii has been shown to possess a substantial slime layer, the basis for attenuation is not likely to be a simple smooth-to-rough variation.


Cold hemagglutinin cross-reactivity with Mycoplasma pneumoniae. Convalescent sera from proven cases of infection with Mycoplasma pneumoniae, and rabbit antisera to M. pneumoniae and to human erythrocyte glycoprotein contained cold hemagglutinins which were reactive only for human erythrocytes. Only the human serum cold agglutinins were inhibited by soluble integral glycoproteins derived from human erythrocyte ghosts by treatment with chloroform-methanol. Rabbit antiserum to chloroform-methanol glycoprotein, as well as to M. pneumoniae, fixed complement with either M. pneumoniae or chloroform-methanol glycoprotein antigens. The findings support the hypothesis that the cold agglutinins elicited by M. pneumoniae infection represent a cross-reaction between determinants common to erythrocyte glycoprotein containing I antigen and the membrane of M. pneumoniae.


Comparison of antigenic heterogeneity of Neisseria gonorrhoeae strains by micro-immunofluorescence and serum bactericidal tests. The antigenic heterogeneity of Neisseria gonorrhoeae strains was assessed by the micro-immunofluorescence (micro-IF) and the serum bactericidal tests. The micro-IF test verified the antigenic heterogeneity of nine strains received from the Center for Disease Control and placed them into immunotypes A and B. The serum bactericidal system also detected different antigenic determinants among the strains. Although the micro-IF and bactericidal assays did not correspond in each instance, the overall pattern of similarities and differences among these gonococcal strains was similar. The micro-IF pattern obtained with mouse antisera was identical to the pattern revealed with guinea pig antisera. Different colony type organisms showed similar sensitivity in the bactericidal test. The micro-IF test is a rapid technique for the immunotyping of N. gonorrhoeae and has the additional advantages of reproducibility and simplicity.


K99 surface antigen of Escherichia coli: antigenic characterization. K99 prepared by acid precipitation hemagglutinated guinea pig erythrocytes, whereas K99 prepared by chromatography on diethylaminoethyl-Sephadex did not. K99 purified by either procedure hemagglutinated horse erythrocytes. K99 prepared by acid precipitation contained a second antigen not presnet in the K99 prepared by chromatography on diethylaminoethyl-Sephadex. This antigen could be detected by immunoprecipitation with some, but not all, sera prepared against K99-positive Escherichia coli strains. It was assumed that this second antigen is not K99 and is responsible for the guinea pig erythrocyte hemagglutination reaction. Furthermore, the second antigen has an isoelectric point of 4.2, which has been reported by Morris and co-workers to be the isoelectric point of K99.


Demonstration of shared and unique immunological determinants in enterotoxins from Vibrio cholerae and Escherichia coli. Immunodiffusion and biological neutralization studies demonstrated that the heat-labile enterotoxin (LT) from Escherichia coli has antigenic determinants in common with each of the isolated subunits (A and B) of the enterotoxin (choleragen) from Vibrio cholerae. Each of the enterotoxins also possesses unique antigenic specificities. Monospecific antiserum to LT was prepared by immunization with antigens derived by immune precipitation of E. coli cell-free supernatant with isolated specific anticholeragenoid antibodies. This antiserum neutralized the biological acitivity of both LT and cholera enterotoxin and recognized antigens of both in immunodiffusion. This antiserum was adsorbed with choleragenoid to remove antibodies directed against the shared "B" immunological determinants. The neutralizing effect of the antiserum on cholera toxin was completely removed, but the neutralizing activity against the E. coli preparations was retained, although somewhat reduced. Antisera to the isolated subunits (A and B) of cholera enterotoxin neutralized the biological activity of cholera enterotoxin and LT. These antisera also recognized the homologous and heterologous antigens in immunodiffusion. Multiple forms or conformations of LT and its components may explain the diversity of the properties which have been reported for it.


Characterization of the cross-reaction between type 19F(19) and 19A(57) pneumococcal capsular polysaccharides: compositional analysis and immunological relation determined with rabbit typing antisera. The immunological relation, physicochemical characteristics, and chemical composition of type 19F(19) and 19A(57) within the cross-reactive group 19 pneumococcal capsular polysaccharides were studied. By using rabbit hyperimmune diagnostic antisera in agglutination, immunodiffusion, quantitative precipitation, and absorption assays, extensive cross-antigenicity and cross-immunogenicity were demonstrable between the disease-associated types 19F(19) and 19A(57). Types 19B(58) and 19C(59), rarely associated with human disease, were extensively cross-reactive with each other but reacted weakly with types 19F(19) and 19A(57). Both types 19F(19) and 19A(57) polysaccharides contained trace amounts of protein and nucleic acid and had comparable molecular sizes as determined by gel filtration. Compositional analysis showed type 19F(19) to contain rhamnose, glucose, N-acetylmannosamine, and a phosphate ester. Type 19A(57) contained these four moieties, and in addition, contained fucose, galactose, and N-acetylglucosamine. Plans for using this information to evaluate current and proposed formulation of multivalent pneumococcal polysaccharide vaccines are discussed.


Secretory immunoglobulin A and G antibodies prevent adhesion of Escherichia coli to human urinary tract epithelial cells. The adhesion of Escherichia coli to human urinary tract epithelial cells was inhibited by commercial gamma globulin, the total immunoglobulin fraction of human breast milk and urine, as well as the isolated immunoglobulin G and secretory immunoglobulin A fractions of urine from patients with acute pyelonephritis. Urinary anti-O6 antibodies reduced the adhesion of several O6 strains. Absorption of antibodies to the lipopolysaccharide of the adhering strain markedly decreased the antiadhesive capacity of all the immunoglobulin preparations, whereas elimination of antibodies to the capsular polysaccharide antigen consistently had a small but not significant effect. When urine was absorbed with whole, live bacteria of the patients' infecting strains, the antiadhesive effect completely disappeared. Absorption with bacteria lacking pili only partially reduced this effect.


Granulocytic function after administration of pepsin treated human gammaglobulin. Pepsin-treated human gammaglobulin, 150 mg/kg body weight, was administered intravenously to 14 healthy volunteers. Before and after the infusion the nitroblue tetrazolium (NBT)-reduction of granulocytes was studied in all and the bactericidial capacity in 12 subjects. An increase of NBT reduction (p less than 0.05) and of bacterial capacity (p less than 0.01) of granulocytes was found after the infusion. The effects may be due to an opsonising action of F (ab')2 components.


Rigid plasma-membrane-derived vesicles, enriched in tumour-associated surface antigens (MLr), occurring in the ascites fluid of a murine leukaemia (GRSL). Extracellular membraneous vesicles of GRSL leukaemia cells were isolated from the ascites fluid bathing the cells in vivo, and from cell washes. Mammary tumour virus-induced antigens (MLr) expressed on the surface of the cells are enriched on these vesicles as compared to plasma membranes isolated from the cell homogenate. The lipid fluidity of the vesicles is much smaller than that of the plasma membranes, and the content of the pertinent lipid parameters, cholesterol and sphingomyelin, are accordingly greatly increased. The extracellular vesicles which are also enriched in sialic acid and 5'-nucleotidase are apparently derived from the plasma membrane, probably at least partly by exfoliation of selected parts or domains of the surface of living cells. An analogy between this shedding of vesicles, the formation of endocytotic vesicles and the budding of viruses is noted; all these processes select or assemble rigid lipid domains of the cell membrane. The possible participation of surface microvilli and sub-lethal autolysis in the process of shedding is discussed.


On the pathogenesis of bleomycin lung toxicity. Bleomycin is potentially capable of inducing a diffuse interstitial fibrosis of the lung, the pathogenesis of which has not yet been elucidated. The authors have demonstrated that after 48 hr of acute treatment, morphological and functional modifications could be seen in type II pneumocytes, which are responsible for surfactant production. This might be the earliest stage of pulmonary damage.


Immunoglobulins in leprosy.

Immunoglobulins in leprosy. Serum immunoglobulins IgG, IgA and IgM were estimated in 22 lepromatous (LL) patients, 28 tuberculoid (TT), 9 borderline tuberculoid (BT), and 8 borderline lepromatous (BL), and compared with 50 normal healthy adult males belonging to a low socio-economic class. Immunoglobulin IgM was invariably significantly raised in TT, BT and LL subgroups of leprosy patients compared to the control but variation among different subgroups was statistically insignificant. Mean serum IgA levels were also raised in TT, BL and LL subgroups but statistically the rise was not significant. In the BT subgroup, significantly low IgA levels were observed both compared to the control and the other leprosy subgroups. Immunoglobulin G levels were significantly raised only in the LL subgroups compared to the control and the other subgroups of leprosy patients. It is proposed that persistently raised gamma globulins and immunoglobulin G, A and M levels observed in lepromatous leprosy patients could be caused by macrophage blockade hindering the suppressor T-cell mediated homeostatic control for immunoglobulins.


Antigenic properties of Bioplast.

Antigenic properties of Bioplast. The influence of heat treatment (at 150 +/- 2 degrees C for 2 hours) on the antigenicity of human Bioplast fibrin powder and human fibrin Bioplast plates was tested in an experimental study. Untreated and heat-treated fibrin powder with incomplete Freund's adjuvant was subcutaneously injected in rabbits. In addition, untreated and heat-treated plates, to which incomplete Freund's adjuvant was added, were subcutaneously implanted. A booster dose was given 8 weeks after the start of the experiment. Blood samples were collected up to 4 weeks after the second injection or implantation, and the sera were tested for precipitating antibodies by the double diffusion technique (OUCHTERLONY'S method). Unlike previous investigators, we found antibodies in sera from rabbits injected or implanted with heat-treated Bioplast products. Thus, the claim that the antigenic properties of all kinds of fibrin can be fully eliminated by heat treatment does not appear to be tenable. In order to avoid the risk of sensitization in man, it is probably advisable to use only human fibrin for the production of Bioplast fibrin powder and Fibrin Bioplast plates for clinical use in human beings.


Comparison of various preparations of Mycobacterium leprae and other mycobacteria by lymphocyte stimulation. Peripheral blood lymphocytes were stimulated in vitro with different mycobacterial antigen preparations and responses were measured as incorporation of tritiated thymidine. Blood donors were 9 patients with lepromatous leprosy, 16 tuberculoid leprosy patients, and 6 healthy individuals with different degrees of exposure to leprosy. The results revealed a good correlation between the responses to M. leprae from human sources and bacilli from armadillos inoculated with human leprosy bacilli, although the latter were less potent stimulators. Responses to BCG and PPD did not correlate with lymphocyte responses to human M. leprae. Acid-fast bacilli grown on LA-3 medium inoculated with human leprosy bacilli stimulated responses which did not correlate with responses to human M. leprae when positive responses were compared, but showed a good correlation when inhibitory effects of the bacilli on thymidine incorporation were studied. The significance of the lymphocyte stimulation test for identification of microbes is discussed.


Heterogeneous staining of rat hepatocyte nuclei by HBFP technique. The non-enzymatic histochemical technique Haematoxylin--Basic Fuchsin--picric acid (HBFP) was studied in fresh-frozen and Carnoy-fixed, paraffin-embedded rat liver sections. The hepatocyte nuclei fell into two populations and showed either a crimson red or purple staining in frozen as well as paraffin sections. The heterogeneous staining of the rat heptocyte nuclei was also present when the tissue sections were stained by Methyl Green--Pyronin stain. The differing nuclear staining was present in the isolated nuclei also. The HBFP technique, therefore, appears potentially useful when applied to liver and other tissues as well.


Graphic representation of the distribution of acetylcholinesterase in cat dorsal root ganglion neurons. Acetylcholinesterase (AChE) activity of primary sensory neurons of the cat has been quantitated and correlated with cell size. Dorsal root ganglia of the fourth and fifth thoracic spinal levels were studied. Frozen longitudinal and cross-sections were collected serially and stained with Cresyl Violet for total cell counts of the ganglia on the left; the average count was 3375 cells. Ganglia from the right were stained for AChE after the method of Karnovsky & Roots (1964) as modified by El Badawi & Schenk (1967), and counterstained with Haematoxylin. Cells were counted in every fourth section and the diameter of each was recorded. AChE-positive cells were classified as brown (B1, B2, B3) and AChE-negative ones as blue (BL). An inverse correlation exists between cell size and AChE activity. High activity was demonstrated in 29% of the cells (B1), moderate activity in 52% (B2), minimal activity in 15% (B3) and 4% were classified as AChE-negative (BL). Small cells with high activity were centrally located in the ganglia whereas large AChE-negative cells were peripherally distributed. Chi-Square analysis revealed that the size of the cell was not independent of the enzyme colour category.


Location of taste buds in intact taste papillae by a selective staining method. Taste buds were found to stain strongly and selectively in intact papillae with highly acidic dyes such as ponceau S. In intact tongues the taste buds in the fungiform, circumvallate and foliate papillae of the cynomolgus monkey and in the fungiform papillae of the rat as well as the taste discs in the fungiform papillae of the frog could be visualized. This method enables a rapid location and counting of taste buds in taste papillae without preparing histological sections. In cynomolgus tongue material fixed in formalin, the dyes penetrate into the buds. In fresh tongues only the taste pore region of the buds stains, which suggests that in vivo taste buds are impenetrable underneath the pore.


Striated myofibrils in anti-myosin stained, isolated chicken gizzard smooth muscle cells. Highly purified chicken gizzard myosin was used to induce antibody production in rabbits. The IgG fraction was separated from the antisera and coupled to fluorescein isothiocyanate (FITC). Specific antibody (AGM) was isolated from the IgG fraction by affinity purification. Comparisons of the specificity of IgG and AGM for chicken smooth muscle myosin revealed a much greater specificity by AGM. Staining with IgG led to an apparent cross-reactivity with guinea pig smooth muscles which was not seen with AGM staining. Therefore, staining of cells for localization of myosin was performed with AGM. Isolated cells were obtained from chicken gizzards either by collagenase digestion or by agitation of glycerinated pieces. Stained cells and cell fragments revealed the presence of myofibrils as structural units with diameters of about 1.0 micrometer. Stained myofibrils occasionally displayed regular banding patterns with a repeating period of about 1.5 +/- 0.2 micrometer. The presence of banded myofibrils in non-cultured cells shows that the organization of the contractile material is similar to that previously reported for cultured cells by Gröschel-Stewart.


Serial serum protein determinations in scoliotic children treated by spine fusion and autotransfusion. In a longitudinal study, serum protein levels were determined by more than 1200 radial immunodiffusion assays in fifty pediatric scoliotic patients whose blood replacement during spine fusion was provided by predeposition autotransfusion. Several serial samples of serum were obtained at intervals from the time of admission through surgery and discharge and they were examined for eight serum proteins: IgG, IgA, IgM, albumin, transferrin, C3c (B1A globulin), beta-lipoprotein, and alpha2-macroglobulin. The results indicated that neither the three phlebotomies performed to obtain predeposition autologuus blood for reinfusion nor the loss of blood during surgery resulted in significant alteration of any of the serum protein levels at the sampling times tested. The greatest decreases in the serum concentrations of all eight proteins were observed during the early postoperative recovery period.