Surface fibrils (fimbriae) of Actinomyces viscosus T14V. Surface antigens of Actinomyces viscosus T14V were released from cell walls by digestion with lysozyme. These were separated by ion-exchange and gel filtration chromatography into fractions rich in carbohydrate or protein. The former contained a polysaccharide high in 6-deoxytalose, along with a peptide fragment from the cell wall. In the protein-rich fractions, material of high molecular weight was present, which contained some carbohydrate and up to 14.3% nitrogen. Aspartic acid, threonine, glutamic acid, lysine, alanine, and glycine were detected in these fractions, along with smaller amounts of 10 other amino acids. Most of the alanine was present as the L isomer and thus was not from peptidoglycan. Electron microscopy of the high-molecular-weight material revealed long fibrils, 3.5 to 4.5 nm in diameter, which resembled those seen on bacterial cells. V-specific antiserum, prepared by absorbing anti-A. viscosus T14V serum with cell walls of the avirulent strain (A. viscosus T14AV), did not react with the 6-deoxytalose polysaccharide but reacted well with isolated fibrils, and this was not inhibited by 6-deoxytalose.


Passive sensitization of human basophils: evidence for heterogeneity in the IgE molecule. Possible functional heterogeneity of human IgE antibody was studied by passively sensitizing human basophils for antigen-induced histamine release. Six sera of known IgE anti-ragweed antigen E and total IgE content were diluted to contain 10 ng of antibody but differed with respect to the ratio of specific/total IgE. The relative ability of the sera to passively sensitize generally reflected the specific/total IgE ratio but one serum was 2.5 times more active than the remainder. This ability did not reflect the IgG antibody level nor was it due to a dialyzable or heat-stable factor. These data indicate functional IgE heterogeneity, probably due to the interaction of the Fc portion of IgE with the basophil receptor.


Enigma of disodium cromoglycate action on mast cells. In rat peritoneal mast cells, disodium cromoglycate showed no inhibitory effect on histamine release values if the cells were preincubated with the drug for 40 min prior to stimulation with antigen, compound 48/80 or ATP. If the drug was added simultaneously with antigen or a low dose of compound 48/80, a repression of histamine release occurred. No such effect was noted if ATP was employed as histamine liberator.


Effect of norepinephrine on in vitro histamine release from rat mast cells. The effect of norepinephrine on compound 48/80-induced histamine release from rat peritoneal mast cells, was studied in an in vitro system. It was found that norepinephrine, within the concentration range 10(-5)--10(-3) M, exerts a significant, dose-related, repressive effect. This effect is greatly potentiated by the beta-antagonist practolol (10(-3) M), throughout the concentration range of 10(-11)--10(-3) M norepinephrine. Methoxyamine, a selective alpha-adrenergic agonist, also represses histamine release in a dose-dependent manner; however, it is not as potent as norepinephrine. The present results would seem to suggest that the repressive effect on histamine release, observed within a low concentration range of norepinephrine, may be due to alpha-adrenoceptor mechanisms.


Chromatographic studies of uremic plasma. In order to characterize the spectrum of possibly toxic retention compounds in uremia we have developed a simple reproducible method of separating fractions of uremic serum by Sephadex G-15 column chromatography. This technique, which requires no prior deproteinisation and is carried out at ambient temperatures, allowed the separation of uremic serum into several well defined fractions. Subsequent thin layer chromatography (TLC) showed that each peak represented a mixture of peptides, and that there were qualitative and quantitative differences between the plasma of normal and uremic patients as well as between patients with acute renal failure and chronic renal failure.


Serological detection of a polyoma-tumor-associated membrane antigen. Syngeneic antisera raised against three polyoma tumors, SEYF-a SESO and SEWE, of strain ABY, A and ASW origin, respectively, contain antibodies directed against multiple specificities. One specificity is cross-reactive for the three polyoma tumors, but appears to be absent from a large variety of other tumors tested. Other, more "public" antigens are shared with a variety of other tumors of viral or non-viral origin. Five different public specificities of this type have been demonstrated and to some extent defined. Since the SEWE ascites tumor, with the shortest passage history, is much less liable to induce antibodies against the public specificities than the long-passaged SEYF-a and SESO, it is likely that the corresponding antigenic determinants arise as a result of passenger virus pick-up during continued passage, or are due to secondary cytogenetic changes.


Testicular biopsy for infertility: a review of sixty-eight cases with a simplified histologic classification of lesions. Sixty-eight testicular biopsies (29 bilateral and 39 unilateral) done for evaluation of oligospermia and azoospermia are reviewed. A simplified classification of the histologic lesions observed is proposed. A brief discussion of the etiology and pathogenesis of some of these lesions is included.


Inhibition of murine sarcoma virus-induced foci formation by cytidine analogues and other drugs chemotherapeutically effective in human malignancies. The present report describes an in vitro culture system using the Kirsten murine sarcoma virus transformation focus assay to evaluate the antiviral activities of 12 commonly used chemotherapeutically effective drugs. Since these drugs are cytotoxic, the plating efficiencies of the cells treated were monitored simultaneously. Using this procedure, Adriamycin, Daunorubicin, Bleomycin Camptothecin, Mithramycin, hydroxyurea, 5-fluorouracil, thioguanine, 9-beta-D-arabinofuranosyladenine, 1-beta-D-arabinofuranosylcytosine (ara-C), azacytidine and cyclocytidine were tested. Of all these compounds tested only the cytidine analogues - cyclocytidine, ara-C and, to a lesser extent, azacytidine - showed selective effect on inhibition of viral foci over cytotoxicity. Studies on the duration of exposure to ara-C indicated that an exposure time of 10-30 h produced the most pronounced effect on the inhibition of foci formation over that of cytotoxicity.


[Cancer en cuirasse, induced by a malignant mixed salivary gland tumor]. A case of cancer en cuirasse is reported, deriving from a metastasizing malignant mixed tumor of the lacrimal gland. Based on histological criteria, the primary benigne prognosis of mixed tumors of lacrimal glands is discussed. According to sensitivity testing to cytostatic agents treatment with 375 mg of bleomycin achieved a recurrence-free interval of eight months. Pathogenesis of secondary effects are discussed (so-drug-induced called bleomycin-lung).


Aldehyde-fuchsin: historical and chemical considerations. The staining mechanisms of Gomori's aldehyde-fuchsin are not yet fully understood. It seemed therefore timely to review the history of this dye class in context with current dye and aldehyde chemistry. In 1861 Lauth treated basic fuchsin with acetaldehyde. This dye became known as Aldehyde Blue, but consisted of violet and blue dyes. Schiff (1866) studied several aldehyde-fuchsins; these compounds contained two molecules of dye and three molecules of aldehyde. Acetaldehyde-fuchsin prepared according to Schiff's directions showed staining properties similar to those of Gomori's aldehyde-fuchsin. This dye class was soon superseded by new dyes more suitable for textile dyeing, and chemical investigations of aldehyde-fuchsins ceased around the turn of the century. Gomori's aldehyde-fuchsin has been regarded as a Schiff base. However, according to chemical data, low molecular aliphatic aldehydes and aromatic amines tend to form condensation products. Correlations of chemical and histochemical observations suggest such processes during aging of dye solutions. Models of dimers and polymers of aldehyde-fuchsin could be built without steric hindrance. The nature of the bonds formed by various components of aldehyde-fuchsin solutions is not clear. However, cystine in proteins, e.g. in basement membranes, apparently does not play a role in the binding of aldehyde-fuchsin by unoxidized Carnoy- or methacarn-fixed sections.


C cell (parafollicular cell) -- immunoreactive thyroglobulin: purification, identification and immunological characterization. In relation to our earlier finding that the thyroglobulin-like material responsible for the cytochemical immunoreaction of C cells was obtained in the peak I fraction of Bio-Gel A-5 m, which included faster sedimenting components of thyroglobulin, the present study has identified the positive reacting component and clarified its immunochemical and immunohistochemical properties. 1. The peak I fraction of dog and hog thyroglobulin was chromatographed on a Bio-Gel A-50 m column. Antiserum to the faster eluted peak I'1 only immunoreacted with C cells. The peak I'1 was then refiltered on Bio-Gel A-150 m column. Antiserum to peak I''1 fraction of both species which was eluted in the first part had high immune specificity for C cells. 2. When 4-30% and 2-16% continuous gradient gels of polyacrylamide were employed, peak I''1 represented a single electrophoretic band corresponding to the component with the largest molecular weight in thyroglobulin. The protein was named C-thyroglobulin. The molecular weight was approximately 2,600,000, four times as large as 19 S, as calculated by relative mobility on the 2-16% gradient gel. 3. In double diffusion tests, anti-peak I''1 antiserum produced two immunoprecipitin lines with its own antigen. The reaction was different from that of anti-19 S antiserum which formed a single line. 4. On immunoperoxidase staining, anti-peak I''1 antiserum reacted to C cells in exactly the same way as anti-calcitonin antiserum. 5. When anti-peak I''1 antiserum was absorbed with calcitonin, the subsequent reaction of the C cells was greatly decreased. The absorption of anti-calcitonin antiserum with increased amounts of peak I''1 abolished the C cell reaction. On the basis of these observations, the possibility that C-thyroglobulin is a biosynthetic precursor of calcitonin exists.


The affinity of concanavalin A and wheat germ agglutinin for components of the muscle spindle. Sections through the soleus muscle of the rat were incubated with concanavalin A (Con A) or wheat germ agglutinin (WGA) conjugated to fluorescein isothiocyanate. Binding of these lectins to structures which comprise the muscle spindle was studied by fluorescence microscopy. The distribution of the lectins was heaviest in the outer capsule of the spindle and at the surface of intrafusal muscle fibres. The periaxial space in the equatorial region of spindles was unlabelled except in the immediate vicinity of the axial bundle. Binding by Con A was more extensive than by WGA, suggesting that more glucopyranoside units are accessible within the muscle spindle than are those of N-acetylglucosamine.


Immunofluorescent and biochemical studies on tyrosine hydroxylase and dopamine-beta-hydroxylase of the bullfrog sciatic nerves. Immunofluorescence specific for tyrosine hydroxylase (TH) or dopamine-beta-hydroxylase (DBH) was accentuated in both proximal and distal segments of the sciatic nerve after ligation. Estimations of the enzyme activities confirmed the above results. Mean axoplasmic flow rates of TH and DBH in bullfrog sciatic nerve were found to be 8 and 123 mm/day, respectively. They were decreased by colchicine or by cold temperatures (4 degrees C).


The inhibitory effect of xanthine derivatives on alkaline phosphatase in the rat brain. Histochemical and biochemical studies were carried out on the inhibition of alkaline phosphatase (A1-P) activity in rat cerebral cortex with various methylxanthine derivatives. The histochemical study revealed that A1-P activity was completely inhibited with 2 mM theophylline or aminophylline, only slightly inhibited with 5 mM of xanthine, and no way inhibited even with 5 mM of diprophylline or caffeine. The biochemical study showed that A1-P activity was markedly inhibited by 1 mM theophylline, to 36% of the control value, and equally markedly by 1 mM aminophylline to 26% of the control value. It was only inhibited to 99% of the control value even by 5 mM diprophylline and conversely slightly activated, to 110% of the control value, by caffeine. The relationship between the pharmacological activities of methylxanthine derivatives and A1-P was studied, and the biological role of A1-P in the central nervous system was also discussed.


Methods of heavy metal electron microscopic histochemistry applied to frog lung surfactant. Four heavy metal staining methods have been applied to frog lung surfactant. Among them, the iodoplatinate method is the only one that almost exclusively visualizes the phospholipid moiety being produced in the lamellated bodies of the pulmonary epithelial cells and forming the backbone of organized structures within the extracellular lining layer. The other three techniques-ruthenium red-osmium tetroxide, osmium tetroxide-ferrocyanide, acidic phosphotungstic acid in chromatic (Rambourg technique)--more or less give electron contrast to glycoproteins and to a lesser extent to the hydrophilic parts of phospholipids. They all show the extracellular lining layer to be a two component system: the content of the lamellar bodies form--when released--membranous configurations, similar to those observed in mammalian lungs; they unfold in an amorphous hypophase, which is apparently secreted by goblet cells of the pulmonary epithelium.


A new selective ultrahistochemical method for the demonstration of calcium using N,N-naphthaloylhydroxylamine. A new, simple, rapid, and highly sensitive and selective method for the ultrahistochemical detection of calcium is described. The reagent N,N-Naphthaloylhydroxylamine (1,8-C10H6CON(ONa)CO) sodium salt was employed in this study for the demonstration of calcium at the subcellular level in relaxed and contracted muscles (smooth muscle of the stomach, thoracic aorta, and myocardial muscle cells) of the rat (in vitro as well as in vivo) and in the human vascular smooth muscle of the aorta with atherosclerotic calcification. Direct evidence of the presence of calcium in the electron-dense reaction products (calcium N,N-Naphthaloylhydroxylamine) is given by X-ray microanalysis of 1,500-2,000 A thick sections. The significance of distributional differences in the localization of calcium in subcellular structures of relaxed and contracted muscles is discussed in relation to the role of calcium in the control of the muscle activity during the contraction-relaxation cycle.


Heat activation of Phycomyces blakesleeanus spores: theromdynamics and effect of alcohols, furfural, and high pressure. The thermodynamic parameters for the heat activation of the sporangiospores of Phycomyces blakesleeanus were determined. For the apparent activation enthalpy (DeltaH(#)) a value of 1,151 kJ/mol was found, whereas a value of 3,644 J./ degrees K.mol was calculated for the apparent activation entropy (DeltaS(#)). n-Alcohols (from methanol to octanol), phenethyl alcohol, and furfural lowered the activation temperature of P. blakesleeanus spores. The heat resistance of the spores was lowered concomitantly. The effect of the alcohols was a linear function of the concentration in the range that could be applied. When the log of the concentration needed to produce an equal shift of the activation temperature was plotted for each alochol against the log of the octanol/water partition coefficient, a straight line was obtained. The free energy of adsorption of the n-alcohols to their active sites was calculated to be -2,487 J/mol of CH(2) groups. Although still inconclusive, this points toward an involvement of protein in the activation process. The effect of phenethyl alcohol was similar to the effect of n-alcohols, but furfural produced a greater shift than would be expected from the value of its partition coefficient. When the heat activation of the spores was performed under high pressure, the activation temperature was raised by 2 to 4 degrees K/1,000 atm. However, with pressures higher than 1,000 atm (1.013 x 10(5) kPa) the activation temperature was lowered until the pressure became lethal (more than 2,500 atm). It is known that membrane phase transition temperatures are shifted upward by about 20 degrees K/1,000 atm and that protein conformational changes are shifted upward by 2 to 6 degrees K/1,000 atm. Consequently, heat activation of fungal spores seems to be triggered by a protein conformational change and not by a membrane phase transition. Activation volumes of -54.1 cm(3)/mol at 38 degrees C and -79.3 cm(2)/mol at 40 degrees C were found for the lowering effect of high pressure on the heat activation temperature.


New maltose Blu mutations in Escherichia coli K-12. Mutations in the genes pgi, pfkA, and ptsG resulted in a maltose Blu phenotype in Escherichia coli K-12, bringing the number of known Blu alleles to six. The Blu phenotype, as visualized by staining with iodine vapor, is a convenient mutant isolation technique.


What makes a difference in learning from media. Media design and selection decisions are often made solely on the basis of how information is to be delivered to the learner. With reference to recent and current research, this paper stresses the need to base such decisions on the compatibility of media with the content to be presented and with the learners. Application of these factors in the design of mediated instruction and the selection of materials is also considered.


What really makes a difference in learning from media, another view. Research has not produced an algorithm for the selection of media formats. More research is required on the effects of presentation techniques. Production decisions are influenced by many considerations.


Developing a new perspective for biomedical Communications. A three-dimensional model is utilized in an attempt to develop a complete picture of the many facets of biomedical communications. This model provides a suitable visual structure from which to explore the complex interactions of individuals, institutions, and activities in defining the field of biomedical communications. The first dimension of the model was developed through a historical overview of the environment of the biomedical communicator and suggests that changing organization goals, client needs, technology, and resources have influenced the evolution of different types of biomedical communicators. The second dimension of the model identifies five major work roles that may be performed by the biomedical communicator of today. The final dimension of the model is developed by a consideration of the major communication methods (illustration, photography, cinematography, television, audiovisual, and computer) of the biomedical communicator.


Radiolabeled sucrose covalently linked to protein. A device for quantifying degradation of plasma proteins catabolized by lysosomal mechanisms. A general method is described for assessing the degradation of proteins metabolized by lysosomal mechanisms. The method depends on the lysosomal trapping of sucrose which is covalently bound to the protein of interest and thus caried into the lysosome with it. The validity of the method was demonstrated in vitro in studies of the catabolism of low density lipoprotein (LDL) by cultured fibroblasts. Sucrose-derivatized LDL was not distinguished from 125I-LDL by fibroblasts, either in terms of surface binding or rate of uptake. 14C from [14C]sucrose-LDL accumulated in the cells as predicted; very little appeared in the trichloroacetic acid-soluble fraction of the medium (2% of total uptake). 14C-labeled metabolites in the cells (modal apparent Mr = 1000-2000) were separated from undegraded LDL by gel filtration. LDL degradation calculated from the 14C metabolites accumulating intracellularly was in excellent agreement with that calculated from paired studies using 125I-LDL. Finally, the validity of the method was demonstrated in vivo using asialofetuin, a protein previously shown to be selectively taken up and degraded by the liver. In principle, the method described should be applicable to the study of the sites of degradation of any of the plasma proteins.


Transient in vivo protein adsorption onto polymeric biomaterials. The adsorption of albumin, gamma-globulin, and fibrinogen was measured on three ex vivo polymeric shunt surfaces [polyvinyl chloride (PVC), Silastic, and segmented polyether urethane (Biomer)] exposed to flowing heparinized, canine blood in vivo. Small amounts of radiolabeled proteins were infused into anesthetized mongrel dogs and the deposition of radioactivity on the walls of femoral arteriovenous shunts was followed with time for two hours following initial blood-polymer contact. Previously, transient in vivo platelet and fibrin deposition onto PVC, Silastic, and Biomer was measured by a similar technique in the absence of anticoagulant. A time-dependent phase of thrombus deposition followed by thromboembolism was observed on the PVC and Silastic shunt surfaces but not on the Biomer surface. In the studies reported here on PVC and Silastic, fibrinogen adsorption was found to predominate initially, though it subsequently desorbed somewhat and was replaced by albumin and gamma-globulin. On Biomer, the adsorption of all three proteins increased with time following initial blood contact and fibrinogen was less prominent initially. The PVC surface was found to become passivated with respect to further thrombogenesis after 60-min exposure to flowing blood, at which time a higher fraction of albumin was present on the surface compared to that at earlier blood contact times. These results indicate that rearrangement of adsorbed protein species occurs with time on polymer surfaces exposed to flowing blood in vivo. Early and predominant fibrinogen adsorption appears to be an important factor in the thrombogenic and embolic events observed on the PVC and Silastic shunt surfaces in vivo.


Epithelioid cell cultures from rat small intestine. Characterization by morphologic and immunologic criteria. Rat small intestinal epithelial cell lines have been established in vitro and subcultured serially for periods up to 6 mo. These cells have an epithelioid morphology, grow as monolayers of closely opposed polygonal cells, and during the logarithmic phase of growth have a population doubling time of 19--22 h. Ultrastructural studies revealed the presence of microvilli, tight junctions, an extensive Golgi complex, and the presence of extracellular amorphous material similar in appearance to isolated basement membrane. These cells exhibit a number of features characteristic of normal cells in culture; namely, a normal rat diploid karyotype, strong density inhibition of growth, lack of growth in soft agar, and a low plating efficiency when seeded at low density. They did not produce tumors when injected in syngeneic animals. Immunochemical studies were performed to determine their origin using antisera prepared against rat small intestinal crypt cell plasma membrane, brush border membrane of villus cells and isolated sucrase-isomaltase complex. Antigenic determinants specific for small intestinal epithelial (crypt and villus) cells were demonstrated on the surface of the epithelioid cells, but they lacked immunological determinants specific for differentiated villus cells. An antiserum specifically staining extracellular material surrounding the cells cultured in vitro demonstrated cross-reactivity to basement membrane in rat intestinal frozen sections. It is concluded that the cultured epithelioid cells have features of undifferentiated small intestinal crypt cells.


Life history of mouse sperm protein. Intratesticular stages. A basic protein fraction, migrating as a single band in acetic acid-urea gel, distinct from histones, was isolated from mouse sperm collected from vasa deferentia and caudae epididymides and was used to immunize female rabbits. The presence of antibodies to the mouse sperm protein (MSP) in the rabbit antisera was demonstrated by a cytoimmunofluorescence procedure using the cells of origin of the antigenic protein, the mature mouse sperm. The specificity of the antisera was verified by fluid and gel precipitation tests and by crossed immunoelectrophoresis. The latter procedure demonstrated the presence of two antigen-antibody systems, consonant with earlier reports that the basic chromosomal protein of mouse sperm is heterogeneous. MSP antigen in situ was recognized by the specific antibodies of the rabbit antisera only after the smear of mature sperm was treated with either of two reducing agents: 2-mercaptoethanol or dithiothreitol. However, when the immunofluorescence procedure was applied to untreated smears of mouse testicular cells, spermatids of all stages from 1 to 14-15 were positive, while spermatocytes, stage 16 spermatids and spermatozoa were negative. After treatment of testes smears with reducing agent, only spermatocytes remained negative. Those observations indicate the following: (a) MSP is immunogenic in a heterologous species; (b) its antigenic sites are detectable in spermatozoa and spermatids of all stages, but not in primary spermatocytes; (c) those antigenic sites become masked at about stage 15 of spermiogenesis and may be unmasked by treatment with a reducing agent. The interpretation is made, therefore, that one or more components of MSP are assembled at the beginning of spermiogenesis and undergo an alteration in the final intratesticular stage of spermatid maturation. That alteration may be presumed to be the formation of disulfide linkages between the cysteine residues.


A cinematographical study of the development of vitally stained Dictyostelium discoideum. We report observations from time-lapse films of the development of Dictyostelium discoideum (Dd) stained with the vital dye neutral red. We have used this dye to enable us to track individual cells, as they move through multicellular tissues in later Dd stages. Our observations lead us to new conclusions about the control of cell movement and cell contact during Dd development, including the tentative conclusion that the aggregation control competences regulate cell behaviour throughout the whole of Dd. development. We are led to specific hypotheses for the mechanisms of later morphogenetic movements and pattern formation.


Steroid receptor content in cytosol from normal and hyperplastic human prostates. Analyses of steroid hormone receptors were performed using a dextran-coated charcoal technique in cytosolic preparations from 40 cases of benign prostatic hyperplasia (BPH) and 10 normal human prostates. Binding data were calculated according to Scathchard. In all BPH specimens, receptors for the synthetic androgen methyltrienolone (MT) were found (mean maximum number of binding sites, 566 fmol/mg DNA; mean Kd, 0.61 nM), and 25 of 28 samples contained progestin [17 alpha, 21-dimethyl-19-nor-4.8-pregnadiene-3,20-dione (R5020)] receptors (mean maximum number of binding sites, 420 fmol/mg DNA; mean Kd, 0.39 nM). No specimen contained glucocorticoid [dexamethasone (9 alpha-fluoro-16 alpha-methyl-11 beta, 17 alpha, 21-trihydroxy-1,4-pregnadiene-3,20-dione); n = 16] or estrogen [17 beta-estradiol or 11 beta-methoxy-17 alpha-ethynyl-17 beta-estradiol (R2858); n = 26] receptors. No correlations were found between receptor content and age of the patients, weight of adenomas, or percentage of different cell types within the specimens. MT receptors were found in all normal prostates, while 5 of the specimens lacked progestin receptors. Estrogen receptors were found in 3 of the normal prostates, whereas none contained glucocorticoid receptors. The ligand specificity of the MT receptor in a normal prostate with minor amounts of progestin receptors was typical of an androgen receptor, and the ligand specificity of the R5020 receptor in a BPH specimen was typical of a progestin receptor. MT and R5020 had approximately the same affinity for the progestin receptor, whereas the relative binding affinity of R5020 for the androgen receptor was below 0.02 compared to that of MT. The androgen receptor was found to be more stable during repeated freezing and thawing than the progestin receptor.


Detection of adenovirus by enzyme-linked immunosorbent assay. A solid-phase direct enzyme-linked immunosorbent assay (ELISA) was developed for the detection of adenovirus antigen in extracts of infected cells by using antihexon serum. Results with simulated clinical specimens consisting of normal nasal wash specimens seeded with varying concentrations of adenovirus type 5 showed that antigen could be detected in extracts of HEp-2 cell cultures inoculated with 10(2.5) 50% tissue culture infective doses (TCID50) and 10(1.5) TCID50 after 2 and 4 days of incubation, respectively. Fifty-three clinical nasal wash specimens containing adenovirus type 5 (stored for 5 years at -70 degrees C) were used to evaluate antigen detection by ELISA in HEp-2 cell extracts and by manifestation of cytopathic effect in human embryonic kidney cells. After 2 days of incubation, 62% were positive by ELISA, whereas none was positive for cytopathic effect. After 4 days of incubation, 76% were ELISA positive and 47% were positive for cytopathic effect. The results according to infectivity titers indicated that clinical specimens containing 10(3.0) TCID50 or greater were all positive by ELISA after 2 days of incubation in HEp-2 cells, and by 4 days all but one specimen containing 10(2.0) TCID50 or greater were ELISA positive. ELISA and immunofluorescent methods for antigen detection were compared using 24 of the 53 clinical specimens containing adenovirus type 5. Nearly equivalent sensitivities were demonstrated. These results suggest that ELISA may provide an alternative method of detecting and identifying adenoviral infections in humans.


Antibodies detectable by counterimmunoelectrophoresis against Bacteroides antigens in serum of patients with inflammatory bowel disease. Heat-extracted antigens from seven species of Bacteroides were used in passive hemagglutination and counterimmunoelectrophoretic tests. Sera from 87 normal persons (group I) and 15 patients with ulcerative colitis (group II) were of low and equal reactivity in passive hemagglutination tests; all positive tests were eliminated by 2-mercaptoethanol reduction of the sera. When these same sera were tested by counterimmunoelectrophoresis with six of the Bacteroides antigens, no significant difference in the percentage of positive reactions was noted. However, using the chi-square test, the seventh antigen, prepared from Bacteroides vulgatus, successfully distinguished the two populations at the 0.025 level. Counterimmunoelectrophoretic tests with the B. vulgatus antigen also provided a means to separate the patients in group II with active disease from those in remission at a P value of 0.01. All the sera from 12 patients with defined Crohn's disease activity indexes reacted with the B. vulgatus antigen in counterimmunoelectrophoretic tests. Reduction and alkylation of patient sera with 2-mercaptoethanol and iodoacetamide removed detectable antibody in 78% of the samples, which suggested a dominant role of immunoglobulin M in the response to Bacteroides antigens.


Indirect immunofluorescence test for serodiagnosis of Legionnaires disease: evidence for serogroup diversity of Legionnaires disease bacterial antigens and for multiple specificity of human antibodies. Evidence obtained by others who used direct immunofluorescence staining to demonstrate serological differences among strains of Legionnaires disease bacterium prompted this study of parameters influencing the ability of the indirect immunofluorescence test to detect human antibodies to Legionnaires disease bacterium. A total of 25 Legionnaires disease bacterium strains, representing four serogroups, were used as immunofluorescence antigens to test selected human sera. The use of diethyl ether in preparing the antigens was discontinued when it was found that titers against ether-killed group 2 (Togus 1-like) antigens were impossible to determine. Instead, heat-killed suspensions of Legionnaires disease bacterium in 0.5% buffered normal chicken yolk sac were used to show the serogroup diversity of the strains and the serogroup specificity of the antibody response of some, but not all, patients with serological evidence of Legionnaires disease. These studies suggest that multiple antigens should be used in serological tests for Legionnaires disease. Furthermore, the fact that some sera contain antibodies that bind equally well to strains of all four serogroups implies that demonstration of a fourfold increase in titer of paired sera when tested with a single antigen should not be interpreted as evidence of infection with a strain of the same serogroup.


Simple, differential staining technique for enumerating rickettsiae in yolk sac, tissue culture extracts, or purified suspensions. A differential staining method employing acridine orange-stained rickettsiae and safranin-stained standardized suspensions of Shigella dysenteriae is described for enumerating rickettsiae harvested during various stages of the growth cycle.


Extractable antigen shared by Peptostreptococcus anaerobius strains. Extracts from several species of gram-positive cocci were prepared by a modification of the Rantz-Randall autoclave method and tested for reactions with rabbit anti-Peptostreptococcus anaerobius (ATCC 27337 and VPI 5737) sera in a capillary precipitin test. Antigen preparations from two reference strains of P. anaerobius (ATCC 27337 and VPI 5737) and six clinical isolates of P. anaerobius reacted with the P. anaerobius antisera. These extracts formed a line of identity by immunodiffusion and displayed at least one precipitin line by immunoelectrophoresis. Absorption of the antisera with either the autoclaved extract or a 10% whole-cell suspension from each of the eight P. anaerobius strains removed the precipitin line(s) observed during immunodiffusion and immunoelectrophoresis. Extracts prepared to other species of Peptococcus, Peptostreptococcus, and Streptococcus did not react with the P. anaerobius antisera in a capillary precipitin test. In addition antisera to Lancefield groups A to G did not react with the extracts from the eight P. anaerobius strains. Preliminary chemical analysis of the extracts from the eight strains showed that they contained approximately 0.2 mg of carbohydrate per ml and 3.6 mg of protein per ml. The rabbit anti-P. anaerobius sera used in this study detected a common antigen(s) shared by strains of P. anaerobius, but did not react with autoclave extracts prepared from other species of gram-positive cocci. This extractable antigen could be used in a capillary precipitin test to rapidly identify P. anaerobius strains isolated in the clinical microbiology laboratory.


Sputum screening by Nomarski interference contrast microscopy. Gram-stained smears of specimens submitted for sputum cultures were compared with direct wet mounts examined by Nomarski interference contrast microscopy (NIM) for enumeration of squamous epithelial cells (EPC) and leukocytes (WBC). The results obtained by the two methods were comparable, but specimens were more rapidly screened and cell types were more readily differentiated by NIM. Specimens submitted for sputum culture over a 3-month period were examined for EPC and WBC by NIM. Twenty-two percent of the specimens had greater than 25 EPC/field or a predominance of EPC (class I), 30% had greater than 25 EPC and greater than 25 WBC/field (class II), and 48% had greater than 25 WBC/field or a predominance of WBC (class III). The clinical relevance of the culture results was determined by reviewing the records of patients whose specimens were included in the study. Class I specimens provided only 30% clinically relevant culture results. Specimens in class II provided useful culture results in 63% of the patients, and 96% of those in class III provided clinically relevant information. The results confirm the value of sputum screening and demonstrate that NIM provides a rapid, simple, and accurate method for sputum screening.


Sulfide silver stainability of a type of bouton in spinal cord motoneuron neuropil: an electron microscopic study with Timm's method for demonstration of heavy metals. Spinal cord motoneuron neuropil (cervical and lumbar enlargements) has been studied at the ultrastructural level after fixation with glutaraldehyde and staining with uranyl acetate and lead citrate. Based on the appearance of the synaptic specializations, different types of boutons were identified and correlated with the classification of bouton types based on osmicated tissue. The sulfide silver method for histochemical demonstration of heavy metals was applied to the same region. The localization of reaction products (silver grains) was predominantly in the terminals. Within the bouton, the grains were mainly in the specialized region of the synaptic contact, and the presynaptic network has also labelled, but to a lesser degree. All stained boutons had the same type of paramembranous synaptic specialization, but not all of the boutons with this type of specialization were stained. The stained boutons are interpreted as a fraction of the 'F' boutons.


A variant of the pyroantimonate technique suitable for localization of calcium in ovarian tissue. Osmium-pyroantimonate solutions for the precipitation of cations are unsuitable for use with delicate mammalian oocytes. A variant of the pyroantimonate technique employing a mixture of pyroantimonate and glutaraldehyde has been found to give successful and repeatable results if a fixation time of 4 hr is used. Calcium-containing antimonate precipitates were localized principally in nuclei, smooth endoplasmic reticulum, Golgi apparatus, mitochondria, and cytoplasmic processes of both oocytes and follicle cells, and along the plasma membrane in small oocytes. Deposits were also concentrated around the periphery of lipid droplets in the follicle cells. The presence of calcium in the precipitates was confirmed by x-ray microprobe analysis.


Immunocytochemical localization of plasminogen activator on porcine kidney cell strain: LLC-PK1 (LP100). An antibody to plasminogen activator (PA) produced by the cultured cells of the pig kidney cell strain LLC-PK1 (LP100) was used to localize PA on the cell's free (unattached) surface. Localization was accomplished by the unlabeled antibody enzyme method (PAP) at the light microscopic level and at the electron microsopic level. Localization was commonly more intense at cell to cell junctions and was associated with blebs and vesiculation in this area. We are proposing that membrane shedding by blebs and vesiculation may be the mechanism of PA release in the LLC-PK1 (LP100) cell strain.


Complement receptor binding of C3b-coated cells treated with C3b inactivator, beta 1H globulin and trypsin. Treatment of 125I-C3b bound to EAC1423b with C3b inactivator (C3bINA) and beta 1H globulin (beta 1H) cleaved the alpha-chain of C3b into 65,000- and 42,000-dalton fragments, both of which remained disulfide-bonded to the intact beta-chain (C3bi). Subsequent treatment with trypsin (0.1 microgram/ml) released 125I into the supernatant and yielded cells coated with a 33,000-dalton fragment of alpha-chain, presumably C3d. These results are in agreement with those obtained by others using fluid phase C3b. C3b-coated cells (EAC1423b) adhered to complement (C) receptors on human erythrocytes, glomeruli, and monocytes. C3bi-coated cells adhered to the receptors on glomeruli and monocytes, but not to those on human erythrocytes. C3d-coated cells adhered only to the monocyte receptors. The findings suggest that the glomerular C receptor recognizes portions of the C3 molecule different from those recognized by either the erythrocyte or monocyte receptors.


Role of surface IgM and IgD in the functional differentiation of human B lymphocytes: effect of papain treatment. Short-term treatment of normal human B lymphocytes with low concentrations of papain resulted in selective and reversible removal of sIgD determinants, whereas HLA and Ia-like antigens, sIgM as well as receptors for E, C3, and FcIgG were unaffected. When studied for their capacity to generate antigen-specific direct PFC, papain-treated (delta-) B cells were highly sensitive to inactivation by even low concentrations of antigen. In addition, these cells were impaired in their ability to cooperate normally with T-helper cells or their humoral product(s).


Demonstration of Fcgamma receptors on human basophil granulocytes. Fcgamma receptors were detected on human basophil granulocytes. The mononuclear cell fraction of human peripheral blood was incubated with heat-aggregated human IgG (HGG) followed by 125I-anti-HGG. Autoradiography of the cells showed that the majority of basophil granulocytes gave a significant number of grains. Basophils were not labeled by preincubation of the same cells with monomeric HGG followed by 125I-anti-HGG. However, the binding of aggregated HGG to basophils was inhibited by the presence of a high concentration of monomeric HGG or its Fc fragment but not by the Fab fragment. Evidence was obtained that Fcgamma receptors are distinct from IgE receptors on the same cells: i) Saturation of basophils with IgE did not affect the binding of aggregated HGG to the cells. ii) Preincubation with and the presence of aggregated HGG failed to affect the binding of 125I-IgE to basophils, or to block passive sensitization of the cells with IgE antibodies. iii) The Fcgamma receptors did not co-cap with IgE receptors. Aggregated HGG failed to induce histamine release from basophils even in the presence of D2O. It was also found that the presence of aggregated HGG on basophils did not modulate IgE-mediated histamine release from the cells.


Ia-bearing bone marrow-cultured macrophages induce antigen-specific helper T cells for antibody synthesis. In bone marrow cell (BMC) cultures supplemented with colony-stimulating factor (CSF), accessory cells develop that are capable of inducing specific helper T cells. These accessory cells become effective after 4 days in culture and can be found not only in the adherent but also in the nonadherent cell population. On the other hand, very few accessory cells with helper cell-inducing capacity are obtained in BMC cultures without CSF. The active BMC-derived cell type has been shown to carry Ia surface antigen, since pretreatment with anti-Ia serum and complement abolished the capacity of these cells to function like macrophages in helper T cell induction. Moreover, the appearance of functional accessory cells in these cultures coincided with the presence of Ia-bearing cells.


Histamine-releasing activity (HRA). I. Production by mitogen- or antigen-stimulated human mononuclear cells. Supernatants from 1- to 2-day cultures of human mononuclear cells induced the release of histamine from basophils. Generation of this histamine-releasing activity (HRA) was stimulated by addition of concanavalin A to the cell cultures. Mononuclear cells were also cultured with SKSD and Candida albicans antigens. Stimulation of HRA production by these antigens was correlated with positive delayed skin reactions. Serial dilutions of supernatants assayed for HRA provided a semiquantitative determination of the level of HRA in mitogen- or antigen-stimulated samples. Antigen increased HRA production when added during the first or second day of culture. Generation of HRA probably requires active protein synthesis, since puromycin was inhibitory, and since preformed HRA could not be recovered from lysed cells. HRA was detected in supernatants after 4 hr, and the effects of antigen stimulation were apparent after 8 hr of culture. Replacement of supernatants with fresh culture medium allowed continued synthesis of substantial quantities of HRA during the second day of culture. A linear correlation was observed between the amount of HRA produced and the mononuclear cell concentration. Our findings provide evidence for the interaction of lymphocytes and basophils via a soluble mediator.


T-T interactions in the induction of antigen-specific human suppressor T lymphocytes in vitro. We intended to investigate whether the suppression of antigen-induced antibody responses in vitro in man by T suppressor cells required contact of T suppressor cells with target cells or whether this effect was mediated by factors released by T suppressor cells. To this end supernatants of antigen-induced T suppressor cells were tested (by a plaque forming cell assay) for their capacity to suppress antibody responses of autologous and allogeneic human peripheral blood lymphocytes. We have shown that supernatants of antigen-specific T suppressor cells, designated as TsF24: a) can suppress an antibody response of autologous but not allogeneic lymphocytes to the inducing antigen; b) are antigen-specific in their effect; and 3) are produced by radiosensitive T cells. Furthermore, the target of the factor is a radiosensitive T cell. These findings taken together indicate that, in the generation of T-effector suppressor cells in man, T-T interactions occur, and in addition, that cellfree factors may be involved in these interactions.


Tumor-specific immunity induced by somatic hybrids. I. Lack of relationship between immunogenicity and tumorigenicity of selected hybrids. Hybrid clones were derived from fusion of TEPC-15 plasmacytoma cells of BALB/c mice with mouse L cells of C3H origin. The morphology, tumorigenicity, and immunogenicity of three representative clones were extensively studied. One clone (LTC-1) showed a morphology intermediate to that of either parental cell and possessed the highest tumorigenic and immunogenic properties. The other two clones displayed a "flat" morphology which differed significantly from that of either parent. One of these two, LTC-4, eventually induced tumors in some (BALB/c X C3H)F1 mice but failed to stimulate protective immunity against TEPC-15 tumor cells in BALB/c mice. The other hybrid clone, LTC-2, has a "very flat" morphology and did not induce tumors, although it was capable of stimulating a significant level of tumor immunity. Histologically, all the tumors induced by hybrid cells were fibrosarcomas rather than plasmacytomas. These results indicate that the morphology of hybrid cells may be correlated with the tumorigenicity as well as the histologic appearance of tumor. In addition, the degree of tumorigenicity of individual hybrid clones does not correspond to their immunogenicity in the host, suggesting that major antigens responsible for immunogenicity may not play an important role in induction of tumors.


Tumor-specific immunity induced by somatic hybrids. II. Elicitation of enhanced immunity against the parent plasmacytoma. Hybrid cells derived from fusion of a BALB/c plasmacytoma (TEPC-15) and L cells (C3H origin) were used to stimulate tumor-specific immunity against the parental plasmacytoma cells. Live hybrid cells induced tumor-specific immunity against TEPC-15 more effectively than mitomycin-treated hybrid or TEPC-15 tumor cells. Adoptive transfer of immunity with spleen cells of mice immunized with hybrid cells was also more effective than that with mitomycin-treated tumor cells. The immunity induced by the hybrid cells was specific to the TEPC-15 tumor because the mice that received immune spleen cells were not protected against challenge with either HOPC-8 or McPC-603 plasmacytomas. T cell populations were primarily responsible for the transfer of specific immunity based on the sensitivity of immune cells to anti-Thy 1.2 and complement. Mice that had established solid tumors were treated with 5 x 10(7) spleen cells to evaluate the therapeutic value of the hybrid-induced immune cells. Tumors in the mice that received immune cells gradually regressed over a 40-day period, whereas tumors on the control mice continued to grow. These results suggest that a rearrangement of tumor-specific antigens on allogeneic hybrid cells can enhance their immunogenicity.


T cell recognition in the mixed lymphocyte response. I. Non-T, radiation-resistant splenic adherent cells are the predominant stimulators in the murine mixed lymphocyte reaction. The ability of subpopulations of murine spleen cells to stimulate a mixed lymphocyte response (MLR) was studied. It was found that T cells (nylon-nonadherent spleen cells) and B cells [G-10 passed and treated with rabbit anti-mouse brain serum (RAMB) and complement (C)] were poor stimulators of an MLR. In contrast, whole spleen cells or B cells plus adherent cells (RAMB +C-treated spleen cells) produced good stimulation. However, a non-T, radiation-resistant splenic adherent cell (SAC) population was up to 20 to 50 times more efficient as a stimulator of an MLR on a per cell basis than an unseparated spleen population. These SAC were shown to express Ia determinants encoded by genes in I-A and I-E/C. These results suggest that Ia+ SAC may be the predominant stimulating cells in spleen cell populations, and the preferential target for T cell recognition in cell interaction events.


Immunologic cross-reaction of stratum corneum basic protein and a keratohyalin granule protein. Antiserum to the stratum corneum basic protein (SCBP) of newborn rat epidermis was used to test for cross-reactive proteins in extracts of skin and in tissue sections. The antibody reacts strongly with epidermal extracts but very poor with dermal extracts. Buffer extracts of epidermis give a reaction of partial identity or identity with the antigen, SCBP. Urea extracts of isolated stratum corneum give a reaction of identity with SCBP. When the proteins of these extracts are separated by SDS polyacrylamide gel electrophoresis, the immunoreactive peak in the 4 M urea extract corresponds to the 49,000 MW SCBP. The immunoreactive peak in the 1 M potassium phosphate extract corresponds to a 52,000 MW protein. This protein is rapidly and transiently labeled after injection of 3H-histidine into newborn rats, in contrast to the SCBP which is labeled after a 5-hr lag. The 52,000 MW protein appears to be an immunologically related precursor of the SCBP. Immunoreactive proteins were localized in tissue sections by the indirect immunoperoxidase method. A strong positive reaction was seen in keratohyalin granules and in the stratum corneum. The reaction of keratohyalin granules corroborates the extraction of a cross-reactive protein by 1 M potassium phosphate, a method for extraction of keratohyalin granules from epidermis. These results are consistent with the hypothesis that a protein of 52,000 MW is present in keratohyalin and is converted to the SCBP (49,000 MW) concomitantly with the conconversion of a granular cell to a stratum corneum cell.


Effects of aging and chronic sun exposure on melanocytes in human skin. Both aging and sun exposure have well-documented effects on the human melanocyte system. Paired biopsies of habitually exposed and nonexposed skin from adjacent anatomic sites were obtained from 8 donors aged 28 to 80 yr in order to study the combined effect of chronic actinic irradiation and chronologic aging. Density of dopa-positive melanocytes was roughly twofold higher in the exposed than in the nonexposed skin at all ages, suggesting an irreversible effect of sun exposure. Melanocyte density declined approximately 6 to 8% of the surviving population per decade in both sites. Dopa-positivity of individual melanocytes was consistently greater in the chronically exposed skin than in the nonexposed skin of the same subject and did not vary with age. These data strengthen and expand earlier observations of age-related melanocyte changes, and explain the apparent paradox of a generalized increase in pigmentation and simultaneous decrease in melanocyte density which frequently accompany advancing age. In addition, the present study suggests that the principal effect of chronic sun exposure on the human pigmentary system is not premature "aging" as currently recognized histologically, but rather activation and/or proliferation of the exposed melanocytes.


Abrogation of cell-mediated immunity by a serum blocking factor isolated from patients with infectious mononucleosis. Lymphocytes from 80% of patients with infectious mononucleosis in this study failed to produce macrophage migration-inhibition factor in response to partially purified early antigen of Epstein-Barr virus or to tetanus toxoid, whereas lymphocytes from normal subjects did produce this lymphokine. Subsequent analysis of serum from the patients with infectious mononucleosis revealed a serum factor that completely abrogated antigen-specific inhibition of migration by human leukocytes as well as lymphocyte blastogenesis. The serum blocking factor was present in sera from 11 (73%) of 15 patients with infectious mononucleos but only in sera from two (13%) of 15 normal subjects. Samples of serum from five of the patients with infectious mononucleosis and five normal subjects were fractionated with use of Sephadex G-200 gel filtration, and the eluants were assayed for several substances known to inhibit cell-mediated immunity. Serum blocking factor activity could be demonstrated only in fractionated sera from patients with infectious mononucleosis. The serum blocking factor is postulated to be either a soluble immune complex or some as yet unidentified immunoregulatory globulin contained in the IgG fraction of human serum.


Antibody to early antigens of varicella-zoster virus during varicella and zoster. IgG antibody to the early antigens of varicella-zoster virus (VZV) was studied during both varicella and zoster by the indirect immunoperoxidase antibody technique. In parallel, complement-fixing, immune-adherence hemagglutinating, IgG, and IgM antibodies to VZV were studied. In both varicella and zoster infections, antibody to the early antigens of VZV appeared three to five days after onset of infection, reached a peak during the second week, and progressively decreased in titer until it disappeared, usually within two months. This antibody usually appeared slightly later than IgG or IgM antibody and grossly correlated with IgM antibody in varicella. In zoster infections, IgM antibody to VZV was not found by the immune-adherence hemagglutination assay at a titer of greater than or equal to 1:4, whereas antibody to the early antigens showed a curve similar to that found in varicella. It is suggested that antibody to the early antigens of VZV be considered as a marker of acute VZV infection, which is associated with a specific and significant IgM antibody response in varicella but not in zoster.


Lipopolysaccharide-derived serotype polysaccharides from Neisseria meningitidis group B. Three immunologically distinct types of polysaccharides have been isolated by diethylaminoethyl (DEAE) chromatography from the lipopolysaccharide extracts of group B Neisseria meningitidis. All types contain a set of common determinants, as well as distinct ones; all of these determinants are detectable by either immunodiffusion or enzyme-linked immunosorbent assay (ELISA). The polysaccharides elute from a Sepharose 4B column in the range of 2-3 x 10(5) daltons and have isoelectric points from 4.2 to 4.3. Their antigenicity is destroyed by oxidation but is unaffected by neuraminidase, lysozyme, or trypsin. One type of polysaccharide cross-reacts with the Gc2 polysaccharide of Neisseria gonorrhoeae in immunodiffusion systems. Chemical analysis indicates that these polysaccharides contain hexoses, hexosamines, 2-keto-3-deoxyoctonate, ethanolamine, and heptose; analysis of amino acids indicates protein contents of less than 0.05%. In contrast to the lipopolysaccharide from which they are derived, these polysaccharides contain no lipid A and less than 0.5% fatty acids. All three types are precipitated by wheat germ agglutinin but not by concanavalin A or fucose-binding protein. Specific inhibition of this precipitation can be achieved with N-acetyl glucosamine. These antigens may be the bases of a lipopolysaccharide-derived typing system for group B N. meningitidis.


Mouse protection test for group B Streptococcus type III. The mouse model has been used previously for the study of protection against challenge with strains of group B Streptococcus types Ia, Ib, and Ic, but investigators have not yet been successful in causing lethal disease when type III strains are inoculated. In this study, six clinical isolates belonging to serotype III were lethal in seven-week-old outbred albino mice; the 50% lethal dose (LD50) for mice tested ranged from 1.8 x 10(4) to 4.6 x 10(6). In contrast, a laboratory prototype strain of serotype III was not lethal. The lethality observed with this protection test depended on both the volume of the inoculum given and the medium in which the streptococci were suspended. LD50 values decreased as the volume of the inoculum, given intraperitoneally, increased, up to a volume of 1.5 ml. Hyperimmune rabbit antiserum to a clinical isolate of type III protected mice against an estimated 90% lethal dose of the isolate. Normal rabbit serum and heterologous antisera, except for one of two antisera to type Ia organisms, were not protective. Absorption of type III antiserum with whole organisms of serotype III or with purified, type III-specific polysaccharide inhibited protection of mice against type III organisms.


The amnion regulates movement of fetally derived alpha-fetoprotein into maternal blood. The present investigation documents that, under normal conditions, most fetally produced AFP reaches the maternal circulation via diffusion across the amnion from amniotic fluid. This has been determined by comparing maternal serum AFP levels with amniotic fluid albumin concentrations in paired samples. The proportionally demonstrated between them indicates a proportional, transamniotic exchange of the two proteins, each originating on opposite sides of the amnion. Albumin is known to reach amniotic fluid by transamniotic diffusion from maternal blood. All amnions restrict AFP movement into maternal serum, but some are distinctly more restrictive than others; in such cases, a relatively greater increase in amniotic fluid AFP concentration would likely have to occur from a fetal lesion before being reflected in maternal serum. Inconsistencies found in several paired samples identify that other variables may also influence passage of AFP to the mother.


Concept of cytostatic therapy in advanced tumours of the head and neck. In cases with inoperable tumours of the head and neck some benefit may still be derived from cytostatics and radiotherapy. One treatment concept is based on intraarterial infusion therapy (bleomycin, methotrexate), while the systemic intravenous application of cytostatic substances (bleomycin, cyclophosphamide) is considered as an alternative of the intra-arterial approach is ruled out. Of the 31 intra-arterially and the 30 intravenously treated patients (all except 2 had squamous cell carcinomas), 64,5% and 20%, respectively, went into remission. This documents the validity of our treatment concept.


Studies on antihistaminic action. II: In vitro and in vivo effect of chlorpheniramine and its analogs on the uptake and retention of 5-hydroxytryptamine by rabbit blood platelets. We have previously shown that the antihistaminic drug, chlorpheniramine maleate (CTM) increased blood histamine levels more rapidly than it did the blood 5-hydroxytryptamine (5-HT) levels in the rabbit (Sankar, Li and Santare, 1974). The present studies show that, while the in vitro addition of CTM to isolated rabbit blood platelets inhibited the platelet uptake of labeled 5-HT-14C, its administration to rabbits increased such in vitro uptake by platelets isolated from recipient animals. This is possibly due to the enhancement of release (depletion) of 5-HT from platelets in vivo. Such depleted platelets, will take up larger amounts of 5-HT on in vitro incubation. Our studies also show increased levels of blood 5-HT within minutes after administration of CTM to rabbits, further indicating that CTM deplets the platelets of their 5-HT on in vivo administration. Our present studies show that D-CTM is more active than brompheniramine in this system, while the fluorine derivative is least active. It is postulated that the antecedent release of histamine and 5-HT from tissues by antihistamines, renders further anaphylactic challenge or hypersensitivity episode less severe.


Maturation of the inhibitory feedback action of oestrogen on follicle-stimulating hormone secretion in the immature female rat: a role for alpha-foetoprotein. The maturation of the inhibitory feedback action of oestrogen on FSH secretion in the immature female rat was studied from 5 days of age until after the first ovulation. To study the role of the oestrogen binding alpha-foetoprotein (AFP) which is present in the blood of young animals, the effects of various doses of oestradiol and of the synthetic oestrogen R2858 (11 beta-methoxy-17-ethynyl-oestradiol), which is not bound by AFP, were compared in ovariectomized rats. A rise in the serum concentration of FSH within 2 days of ovariectomy was first observed in rats ovariectomized at 8 days of age. Between 8 and 28 days of age the rise in FSH after ovariectomy could be prevented by oestrogen injections in such a way that the resulting FSH concentration amounted to 50% of that in ovariectomized control rats. This was achieved with a constant dose of 0.00015 microgram R2858/100 g body weight, whereas the dose of oestradiol needed decreased from 0.05 to 0.01 microgram/100 g body weight indicating an increased sensitivity to the feedback action of oestradiol. After day 28, sensitivity to the feedback action of both R2858 and oestradiol decreased progressively up to the time of the first ovulation. In contrast to results at earlier ages, none of the doses of either oestrogen was capable of maintaining near-physiological concentrations of FSH after 20 days of age. It is concluded that the apparent increase in sensitivity to the feedback action of oestradiol occurring before 28 days of age reflects the disappearance of AFP from the blood, whereas the subsequent decrease in sensitivity is independent of AFP. Moreover, it is concluded that up to about 20 days of age oestradiol could be, though not necessarily is, the sole ovarian factor involved in regulating FSH secretion, whereas at later ages additional steroids and/or factors must be involved.


In vitro model for natural tolerance to self-antigens. Inhibition of the development of surface-immunoglobulin-negative lymphocytes into T-dependent responsive B cells by antigen. Neonatal and adult splenic cell suspensions were labeled with fluorescein isothiocynate-anti-Ig and fractionated into surface-immunoglobulin- (s-Ig) positive and s-Ig-negative subpopulations by the fluorescence-activated cell sorter. The subpopulations were then tested by splenic focus assay for both frequency and tolerance susceptibility of clonable 2,4,-dinitrophenol (DNP) precursors. It was shown that both adult, and neonatal, s-Ig-negative subsets contained clonable DNP-specific B-cell precursors. However, because these precursors result in fewer clones secreting IgG, they appeared to be less mature than the s-Ig-positive precursors. In the absence of helper T cells, it was found that exposure of s-Ig-negative lymphocytes to tolerogen during the process in which they were acquiring surface receptors resulted in nearly total abrogation of potential DNP clones. This finding provides compelling evidence for clonal abortion.


Ia determinants on stimulated human T lymphocytes. Occurrence on mitogen- and antigen-activated T cells. Human T-cell blasts were generated by stimulation with mitogens and antigens. A proportion of these blasts expressed Ia antigens detectable by immunofluorescence with both allo- and hetero-antiserums. The maximal expression of Ia antigens was delayed and usually occurred after the peak of blastogenesis. Among the three mitogens used, pokeweed mitogen (PWM) was most effective in giving a high percentage and intense Ia staining of T-cell blasts. Phytohemagglutinin and concanavalin A blasts gave weaker and lower percentages of Ia staining. Activation by alloantigens and soluble antigens such as tetanus toxoid and purified protein derivative resulted in Ia expression on T cells comparable to PWM stimulation. Depletion of Ia+ cells from freshly isolated T cells with anti-Ia and complement decreased subsequent Ia expression, suggesting that a proportion of Ia+ blasts were derived from Ia-bearing peripheral blood T cells. When the specificities of the Ia antigens on T-cell blasts were examined with alloantiserums, it was evident that the T blasts expressed similar HLA-DR determinants to those on B cells from the same donor; occasional minor differences between stimulated T cells and autologous B-cell lines or fresh B cells were encountered.


Fine specificity of regulatory T cells. II. Suppressor and helper T cells are induced by different regions of hen egg-white lysozyme in a genetically nonresponder mouse strain. We have examined the ability of two purified peptide fragments derived from hen (chicken) egg-white lysozyme (HEL); N-terminal, Co-terminal peptide (a.a. 1--17:cys 6--cys 127:120--129) and mixed disulfide LII peptide (LII) (a.a. 13--105) to induce antigen-specific suppression or help in B10 (H-2b) nonresponder and B10.A (H-2a) responder mice. An anti-HEL primary in vitro antibody response can be obtained in either strain by stimulation with HEL coupled to erythrocytes (RBC). Preimmunization with HEL-complete Freund's adjuvant-(CFA) or N-C-CFA-induced suppression of the anti-HEL PFC response to HEL-RBC in spleen cell cultures from B10 mice, whereas helper activity was demonstrated in cultures from B10.A mice similarly immunized. LII-CFA priming elicited helper cells in both C57BL/10 Sn (B10) and B10.A/SgSn (B10.A) mice. The genetic nonresponsiveness of B10 mice to HEL can therefore be attributed to the activation of suppressor T cells by a limited portion of the molecule (e.g., N-C) which prevent the potential response directed against other epitopes on the same molecule (e.g., LII). One manifestation of major histocompatibility complex gene activity appears to be the intramolecular selection of different antigenic determinants leading to activation of functionally different T-cell subpopulations.


Release of prolactin and LH and histamine-containing cells in brain. The effects of histamine released from mast cells by the compound 48/80, a degranulating agent, on plasma prolactin and LH levels, were examined in rats. Compound 48/80 given systemically (s.c.) at a dose of 2 mg/100 g b.w. significantly increased at 15 and 60 min after injection, both prolactin and LH release in ovariectomized, steroid-primed rats. Prolactin but not LH was also augmented in the plasma of male rats following s.c. administration of compound 48/80. There were no modifications of plasma prolactin levels in male rats at 15, 30 or 60 min after intraventricular injection of 50 microgram compound 48/80. Results suggest that the hormone effects were due to histamine released from median eminence mast cells after systemic injection of compound 48/80.


Taurine in the developing rabbit visual system: changes in concentration and axonal transport including a comparison with axonally transported proteins. [35S]Taurine injected intravitreally into rabbits was transported axonally to the optic nerve terminals. Considerably more [35S]taurine was transported in young rabbits than in mature rabbits. The time course of taurine transport did not parallel that of proteins labeled with [3H]proline in the same system. The concentration of taurine in all components of the visual system, except retina, was greater in young animals than in mature animals, and was especially high in optic nerve. The possible functions of the high concentrations of taurine and the greater amount of axonally transported taurine in developing mammalian CNS are discussed.


Muscle fibrillation caused by cytochalasin-B applied to the motor nerve. Cytochalasin-B, a drug known to interfere with axoplasmic transport, evoked fibrillary potentials in the geniohyoid muscle when applied to its motor nerve. Despite this denervation-like effect, neuromuscular transmission remained normal. Some contractile characteristics of the muscle were studied. It was found that contraction time, isometric twitch tension, and half-relaxation time were not altered by the drug treatment. The present findings show that neurogenic molecular factors conveyed by axoplasmic transport to the nerve terminal are involved in the regulation of some muscle membrane characteristics but do not modify the muscle contractile features.


Striato-nigral astrocytic melanization.

Striato-nigral astrocytic melanization. Abnormal pigmentation was found in the striatum, pallidum and substantia nigra of two unrelated patients. Both had suffered from debilitating joint disease; there was no evidence of a progressive neurologic disorder. The pigment was localized in astrocytes and had the histochemical and electron microscopic features of melanin. The tissue changes suggest an anomaly of catecholamine metabolism either endogenous or induced by a still unknown exogenous factor.


Ischemic lesions of the hippocampus and their relation to Ammon's horn sclerosis. A neuropathological study of two cases and a comparison to the vascular anatomy. Two adult brains with small ischemic lesions in the hippocampus, due to impairment of the supply from the posterior cerebral artery, are presented. The first case corresponds to what is described in the literature as "incisural sclerosis" and shows no difference in vulnerability between the Sommer and the Spielmeyer sector. In the second case the hippocampal lesion is due to an embolic occlusion of the posterior cerebral artery and consists of selective necrosis of the subiculum, the Sommer sector and part of the endfolium of the pyramidal layer, the Spielmeyer sector remaining noninfarcted. The postmortem angiograms of 12l hippocampi of adults, as well as full term born and premature infants, show that the h1 and h2 sectors and part of the h3 sector of the hippocampus are supplied by the same "sulcus" arteries. Although there is a selective vulnerability to ischemia in some sectors of the hippocampus, which is typical for Ammon's horn sclerosis, this cannot be explained by a difference of arterial supply or by compression of arteries during the process of birth.


Nerve conduction in the Guillain-Barré-Strohl syndrome. Fifty cases of the Guillain-Barré-Strohl syndrome were investigated clinically and electrophysiologically--20 in the acute phase, and 30, as a matter of followup, many years after. The sural nerve was biopsied in six cases. There was no evident correlation between clinical symptoms and slowing of motor and sensory conduction. Nerve conduction velocity became slower after the beginning of clinical improvement. The electrophysiological abnormalities concerned both sensory and motor fibers despite the frequent absence of clinical sensory manifestations. The so-called long nerves were involved earlier and more markedly than the so-called short nerves. Conduction velocity and distal latency were equally affected. A slight electrophysiological defect was noticeable even many years after the acute phase of the syndrome, in completely symptoms free patients. Some correlation existed between conduction velocity changes and histological findings.


Effect of furosemide (lasix) on acute severe experimental cerebral edema. The effect of furosemide (Lasix) therapy on a standardized experimental cerebral edema, induced in rats by applying a cooling stamp to the right side of the skull over the right coronal suture by means of a stereotactic instrument, was examined. The hemispherically separated water and electrolyte contents of the brain were analyzed after 24 h. Following furosemide therapy, the behavior of these edema parameters was compared statistically with dexamethasone, glycerol and albumin. An increase of the water and sodium content, and a decrease of potassium was observed 24 h after the trauma, especially in the right hemisphere. Furosemide did not improve either the water content or the electrolyte balance. By contrast, the administration of dexamethasone, glycerol and albumin was followed by a significant improvement of the edema. In experiments with cats, the course of the edema and the effect of furosemide on the cold brain injury of the right hemisphere were observed by measuring the intracranial pressure (ICP) values, and by continuous monitoring of the EEG. The ventricular CSF pressure and epidural pressures were also recorded. The electrical brain activity was continuously compared with the course of the ICP by means of computer analysis. In addition, the blood osmolality and diuresis were monitored. The ICP increased rapidly after the trauma, establishing considerable pressure gradients, and the EEG power intensities decreased markedly on the right side. Histologically, there was an extended edema of the white matter of both hemispheres. The ICP was not lowered by single injections or high dose infusions of furosemide, and the EEG power intensities also did not improve. Infusions of large volumes of furosemide even resulted in an increase of ICP, but infusion of 40% sorbitol effected a rapid decrease of ICP and EEG recovery over the left hemisphere. Sorbitol infusion also caused a marked rise in the blood osmolality, whereas furosemide had no such effect. The results raise considerable doubts as to the propriety of the exclusive use of furosemide for cases of acute cerebral edema with raised ICP. The diuretic effect is insufficient to establish an osmotic gradient, and its general dehydrating effect does not acutely influence the ICP. The absence of effect on the experimental tissue edema would not appear to commend furosemide as basic therapy for cases of traumatic cerebral edema.


The response of focal ischemic cerebral edema to dexamethasone. Twenty-four h after permanent occlusion of the middle cerebral artery (MCA) in the cat, the hemispheric swelling due to edema is markedly reduced under treatment with large doses of dexamethasone than is the case with the untreated group. The increase of regional water and sodium content in the MCA territory is less in the dexamethasone treated group, whereas the potassium changes in the ischemic tissue showed only small differences between the two groups. The potassium content of the non-ischemic tissue is slightly increased in the dexamethasone treated animals when comparing with the untreated group. RISA activity in the tissue is increased in the grey and the white matter of both groups. The less marked RISA-131 activity in the cortical grey matter of the treated animals indicates blood-brain barrier damage of a smaller degree due to dexamethasone. These findings indicate a beneficial effect of dexamethasone on local ischemic edema. Regarding our results and the pharmacokinetics of this steroid the dexamethasone loading of a patient has to be in the range of about 100 mg per day for the adult, and has to be started immediately after the onset of a stroke.


Epileptic laughter: electroclinical and cinefilm report of a case. A woman, aged 30, experienced attacks of pathological laughter which began during the first months of life; they meet the qualifying criteria of epileptic laughter. Several attacks were recorded by polygraphy and cinefilm. The attack begins with a feeling of unreality associated with a forced, involuntary smile progressing to full laughter. The patient tried to mask the laughter which was not accompanied by euphoria but was followed by loss of consciousness and automatisms. Only during the latter phase of the previously normal EEG did an ictal discharge appear over the right hemisphere. On the basis of the electroclinical pattern and of a review of the literature, the seizure is tentatively explained in terms of a progressive ictal involvement of temporodiencephalic structures.


5-Azacytidine hydrolysis kinetics measured by high-pressure liquid chromatography and 13C-NMR spectroscopy. Hydrolysis of 5-azacytidine, an experimental anticancer drug, in aqueous buffers was measured using a high-pressure liquid chromatographic (HPLC) procedure and a 13C-NMR method. The former utilized a 17.5-micron Aminex A-6 strong cation-exchanger column eluted with 0.4 M, pH 4.6 ammonium formate buffer at a flow rate of 0.4 ml/min. The hydrolysis sequence as well as the existence of a labile intermediate, N-formylguanylribosylurea, was unequivocally established using 6-13C-5-azacytidine and NMR spectral techniques. A reversible ring opening step to the N-formylguanylribosylurea with an equilibrium constant of 0.58 +/- 0.03 between pH 5.6 and 8.5, followed by an irreversible formation of guanylribosylurea, was found by HPLC. The data confirm previous assumptions on the hydrolytic kinetics. The pH dependency of hydrolysis was examined, and the hydrolysis profile gave a normal V shape with the most stable pH at 7.0. Rather stable intravenous dosage forms can be formulated.


Stabilization of 5-azacytidine by nucleophilic addition of bisulfite ion. 5-Azacytidine (I) stability was increased approximately 10-fold over its stability in water or lactated Ringer injection by the addition of excess sodium bisulfite and the maintenance of pH approximately 2.5. The increased stability in the presence of bisulfite at pH 2.5 was attributed to the addition of bisulfite across the 5-6 protonated imine bond of I, which prevented the hydrolytic attack on this labile double bond. However, above pH 4, bisulfite increased I degradation. At higher pH, the compound was no longer protonated and bisulfite did not form the stable addition product. The addition compound quickly decomposed above pH 6 to give back the parent compound and, thus, acted as a I prodrug. The intact drug remaining was assayed by high-pressure liquid chromatography (HPLC), and the reversibility of the bisulfite-I addition product above pH 6 was demonstrated by UV spectrophotometry and HPLC. The potential utility of the bisulfite-I addition product as a I prodrug in parenteral and possible oral dosage forms is discussed.


Neural control of skeletal muscle cholinesterase: a study using organ-cultured rat muscle. 1. It has been proposed that the influence of innervation on the cholinesterase activity (ChE) of skeletal muscle and on end-plate ChE in particular is mediated by trophic substance(s) moved by axonal transport and released from nerve. We have tested this hypothesis using rat extensor digitorum longus (e.d.l.) and diaphragm muscles denervated in vitro for several days and then maintained in organ culture to assay putative trophic substance(s). 2. The cholinesterase activity (ChE) of rat extensor digitorum longus (e.d.l.) muscles decreased dramatically after 5 days of denervation in vivo as previously reported. The ChE of rat e.d.l. muscles denervated in vivo for 3 days and then maintained in organ culture for 2 days was essentially identical to that of muscles denervated 5 days in vivo. 3. The ChE OF E.D.L. MUSCLES DENERVATED IN VIVO FOR 3 DAYS AND THEN MAINTAINED FOR 2 DAYS IN CULTURE MEDIUM SUPPLEMENTED WITH SCIATIC NERVE OR INNERVATED MUSCLE EXTRACT WAS SIGNIFICANTLY HIGHER THAN THAT OF MUSCLES DENERVATED IN VIVO FOR 5 DAYS OR DENERVATED IN VIVO FOR 3 DAYS AND THEN CULTURED FOR 2 DAYS IN CULTURE MEDIUM ALONE. Supplementing the culture medium with brain or spinal cord extract also significantly increased the ChE of organ-cultured e.d.l. muscles. 4. Supplementing the culture medium with liver or spleen extract or with the extract of muscle denervated for 3--7 days in vivo before extraction did not increase the ChE or organ-cultured e.d.l. muscles. 5. The effect of muscle extract on the ChE of organ-cultured e.d.l. muscles was dose dependent and occurred gradually reaching a maximum after approximately 24 h of culture. 6. Substance(s) which increased the ChE of organ-cultured e.d.l. muscles were found to accumulate in transected sciatic nerve in the region just proximal to the site of transection where substances moved by axonal transport are known to accumulate. 7. Media conditioned with neurally stimulated e.d.l. or diaphragm muscles caused a substantial and highly significant increase in the ChE of e.d.l. or diaphragm muscles denervated in vivo and then maintained in organ culture. Media conditioned in the same way with unstimulated muscles did not increase the ChE OF ORGAN-CULTURED MUSCLES. 8. The active substance(s) released by neural stimulation continued to be released when muscle contraction was blocked by adding D-tubocurarine to the medium during conditioning but the release of these substance(s) was significantly reduced when magnesium (10mM) was added to the medium during conditioning. 9 The substance(s) released by neural stimulation selectively increased ChE in the end-plate region. In diaphragm segments denervated in vivo and then maintained in medium conditioned with neurally stimulated muscle, there was a 102% increase in end-plate ChE but no detectable increase in background ChE. 10...


Synthesis of 3,4-dihydro-4-oxothieno[2,3-d]pyrimidine-2-carboxylates, a new series of orally active antiallergy agents. A series of novel 3,4-dihydro-4-oxothieno[2,3-d]pyrimidine-2-carboxylic acid derivatives has been prepared and tested for antiallergenic activity. Members of the series, including both carboxylic acid salts and esters, have been found to exhibit oral activity in the rat passive cutaneous anaphylaxis (PCA) test. Activity is optimized by H or CH3 substitution at the 5 position and lower alkyl groups at the 6 position. Ethyl 6-ethyl-3,4-dihydro-4-oxothieno-[2,3-d]pyrimidine-2-carboxylate and 3,4-dihydro-5-methyl-6-(2-methylpropyl)-4-oxothieno[2,3-d]pyrimidine-2-carboxylic acid dipotassium salt were the most potent of the esters and salts, respectively. Such compounds have been shown to have a duration of action of up to 4 h in the PCA test and to inhibit both histamine release from rat peritoneal mast cells in vitro and allergen-induced bronchospasm in the rat lung.


Pyranenamines: a new series of antiallergic compounds. Condensation of 3,5-diacylpyrantriones with various aromatic amines gave a new class of potent, orally active, antiallergic compounds, the 3-[(arylamino)ethylidene]-5-acylpyrantriones, hereafter referred to as pyranenamines, as evaluated not only in the traditional rat passive cutaneous anaphylaxis (PCA) assay but also in the in vitro fragmented rat and primate lung assay. Potencies in the PCA system, when measured intravenously, reached a maximum ID50 of 0.9 mu/kg (1000 times more potent than disodium chromoglycate) with 5-acetyl-4-hydroxy-3-[1-[(3,5-bis-glyceramoylphenyl)amino]ethylidene]-2H-pyran-2,6(3H)-dione (100), as predicted by structure-activity relationship (SAR) analysis. Potencies in the iv PCA system correlated well with potencies in the in vitro rat lung system but not with potencies in the oral PCA system or the in vitro primate lung system. Several compounds had good oral potency, and one analogue, 3-acetyl-4-hydroxy-3-[1-[3-amino-4-hydroxyphenyl)amino]ethylidene]-2H-pyran-2,6(3H)-dione hydrochloride (78), reached an oral ID50 of less than 1 mg/kg and was better than 10 times more effective than disodium chromoglycate at inhibiting the release of histamine and slow-reacting substance of anaphylaxis in the fragmented primate lung assay.


Antiallergy activity of substituted 11-oxo-11 H-pyrido[2,1-b]quinazoline-8-carboxylic acids. A series of substituted 11-oxo-11H-pyrido[2,1-b]quinazoline-8-carboxylic acids were prepared and evaluated as antiallergy agents. Several analogues were orally active. 2-Methyl-11-oxo-11H-pyrido[2,1-b]quinoazoline-8-carboxylic acid (6) was superior to cromolyn sodium and doxantrazole orally and intravenously in the rat PCA test and a rat allergic bronchospasm model.


A technique for investigating the antigenicity of cultured rheumatoid synovial cells. Antisera to rheumatoid and nonrheumatoid synovial cells were prepared by: intraarticular injection of rabbits with these cells, subsequent synovectomy, and autologous immunization of each animal with the cultured rabbit synovial cells, or homogenized whole rabbit synovial membrane. Resultant antisera were studied by immunofluorescence and 51chromium cytotoxicity tests against other cultured rheumatoid and nonrheumatoid cells. Activity of positive sera could be removed by absorption with nonrheumatoid cells, except for one antiserum which appeared to produce immunofluorescent staining specific for rheumatoid cells.


A modified levanol fast cyanine 5RN staining method for myoepithelial cells. The specificity of the tannic acid--phosphomolybdic acid--Levanol Fast Cyanine 5RN procedure for myosin-like proteins in myoepithelial cells is hampered by the affinity of the dye for other structures. The selectivity of myoepithelial cell staining is enhanced by oxidation of sections, nuclear staining by Safranin-O, and differentiation with Tartrazine.


The clinical significance of autoantibodies to a soluble cytoplasmic antigen in systemic lupus erythematosus and other connective tissue diseases. Seventy-two patients were studied whose sera contained antibodies to a cytoplasmic antigen, RO. The majority had clinical features typical of systemic lupus erythematosus and there appeared to be an overlap with Sjögren's syndrome. Of interest is that certain patients with lupus respond predominantly to cytoplasmic antigens and in some cases were ANA negative. Demonstration of this system is important in enlarging the serologic spectrum characteristic of SLE patients, and also may be an important marker for systemic disease in patients with predominantly cutaneous involvement.


Role of radiotherapy in the management of breast cancer. There is no subject as controversial as the treatment of early breast cancer in spite of 90,000 new cases per year and more than 30,000 deaths per year in the USA alone. Radical mastectomy has been in practice for over 80 years. Radiotherapy, alone, was used 50 years ago. Today, however, we are unable to state which is the method of choice in the management of early breast cancer. Radiotherapy did not become popular as the primary treatment modality during the radium and orthovoltage era because of technical problems. These technical problems have been overcome during the afterloading and supervoltage era. Unfortunately, we do not have a large series in radiotherapy to compare with those of 80 years of radical mastectomies. Until such time, radiotherapy will play an adjuvant and palliative role in the management of breast cancer patients.


Patterns of inflammation in prostatic hyperplasia: a histologic and bacteriologic study. In a series of 162 cases of surgically resected hyperplastic prostates the incidence of inflammation is 98.1%. Six morphologic patterns of inflammation are described: 1) segregated glandular inflammation, 2) periglandular inflammation, 3) diffuse stromal inflammation, 4) isolated stromal lymphoid nodules, 5) acute necrotizing inflammation and 6) focal granulomatous inflammation. The most common pattern, segregated glandular inflammation, is characterized by intraluminal neutrophils and foamy macrophages and by chronic inflammatory cells in the surrounding stroma. No significant morphological differences are found among groups of cases with positive and negative evidence by culture of bacterial prostatic infection. Quantitative but not qualitative morphologic differences are found between cases of gram-negative infections and infection by gram-positive organisms that often are considered non-pathogens.


Falsely positive specific human chorionic gonadotropin assays in patients with testicular tumors: conversion to negative with testosterone administration. We report false elevations of serum human chorionic gonadotropin levels in 4 patients with testicular germ cell tumors. Elevated circulating luteinizing hormone levels that resulted from unilateral orchiectomy were responsible for the falsely positive human chorionic gonadotropin activity measured in commercial radioimmunoassays. In 3 patients tested aliquots of serum evaluated in reliable human chorionic gonadotropin assay systems revealed no elevation. We administered testosterone to 3 patients who had elevations of luteinizing hormone and human chorionic gonadotropin levels. Luteinizing hormone was suppress-d to normal levels 1 week later in 2 patients and in 1 it was diminished but still slightly elevated. In all 3 cases the falsely positive human chorionic gonadotropin results converted to negative. Recognition of falsely positive elevations of human chorionic gonadotropin can spare patients unnecessary operations and/or chemotherapy.


Specific binding of prolactin by the prostate gland of the rat and man. Specific binding sites for 125iodine-prolactin are present in membrane particles obtained from the rat ventral prostate, human benign prostatic hyperplasia and prostatic adenocarcinoma. In the ventral prostate glands of young rats (1 to 4 months old) specific binding of 125iodine-prolactin is higher than in older animals (greater than 8 months old). Subcellular distribution studies revealed that specific 125iodine-prolactin binding activity is associated primarily with the 15,000 and 100,000 g particulate membrane fractions of the rat ventral prostate and human prostate glands. In rats between 2 and 4 months old significant increases in the prolactin binding activity in the 100,000 g membrane fraction of the ventral prostate are observed to occur without concomitant increases in prolactin binding activity in the 15,000 g fraction. The level of specific 125iodine-prolactin binding activity present in the human prostate gland is lower than that observed in the rat ventral prostate gland. Localization of specific prolactin binding sites in the rat ventral psotate and the human prostate gland suggests that prolactin could influence the function of these tissues directly.


Techniques for evaluating humoral and cell-mediated immunity in mule deer fawns (Odocoileus hemionus). Twenty mule deer fawns (Odocoileus hemionus) were removed from their dams 48 h after birth, and hand-reared. Methods for monitoring their immune capability are described. Passive humoral immunity was determined by serum protein electrophoresis. Active humoral immunity following Clostridium toxoid vaccination was determined by immunodiffusion. Cell-mediated immunity was assayed using contact sensitization to 1-nitro, 2,4-dichlorobenzene (DNCB).


Sensitization of leukocytes of cancer patients against fetal antigens: leukocyte migration studies. Hypertonic extracts from human fetuses (10--22 wk of gestation) were used to test the sensitizaton of leukocytes from cancer patients against fetal antigens in a direct, microcapillary tube assay system. Leukocytes were simultaneously exposed to a panel of allogeneic tumor extracts and a panel of fetal extracts. Leukocytes from 24 gastric cancer patients, 43 colorectal cancer patients, and 13 lung cancer patients were assayed with extracts obtained from gastric, colorectal, and oat cell carcinomas, respectively, and these extracts were also used with leukocytes from 41 patients bearing tumors of various other organs. Significant migration inhibition by tumor extracts was observed in 81.6% of the tests with gastric cancer, 67.4% of the tests with colorectal cancer, 69.0% of the tests with lung cancer, and 51.2% of the tests with other types of cancer. With fetal extracts, significant migration inhibition occurred in 58.3, 58.7, 59.6, and 54.9% of the tests, respectively. Reactivity against fetal extracts did not depend on the gestation age of the fetuses used for extraction. The conclusion was reached that the leukocytes of most of the cancer patients were sensitized against substances contained in fetal extracts irrespective of the type of tumor of the leukocyte donor. The cross-reactivity pattern suggested that 3-M KCl extracts of whole human fetuses contained a complex mixture of specificities related to the various fetal organs and tissues, which may have represented counterparts to most of the tumor-associated specificities.


Production and characterization of xenogeneic antisera to a human renal cell carcinoma-associated antigen. Antisera to human renal cell carcinomas were produced by the immunization of goats and rabbits with dissociated tumor cells and/or tumor homogenates from single donors. After absorptions with human red blood cells and homogenates of human liver, lung, spleen, and heart, all the immune sera reacted on immunofluorescence with the brush border of the proximal convoluted tubules of adult and fetal human kidneys and with the proximal convoluted tubular epithelia of rabbits, guinea pigs, rats, and mice. After further absorption with pooled normal human kidney homogenates, the immune sera on immunofluorescence showed cytoplasmic staining of smears and sections of 21 of the 22 human renal cell carcinomas tested. These sera did not show any staining of normal adult human tissues including normal kidney adjacent to the carcinomas, perirenal fibroblasts and peripheral blood leukocytes from the patients, human fetal kidneys, transplanted renal adenocarcinoma of BALB/c mice, and several human tumors tested, i.e., transitional cell carcinoma of the bladder, adenocarcinomas of the breast and colon, squamous cell carcinoma of the lungs, malignant lymphoma, and melanoma. Autoradiography of tissue sections with 131I-labeled antitumor globulins revealed greater localization of radioactivity in tumors than in adjacent normal kidney. Membrane immunofluorescence with the immune sera rendered tumor-specific after appropriate absorptions revealed tumor-associated antigens on the surfaces of all 5 human renal carcinoma cell lines tested.


Immunospecificity of nuclear nonhistone protein-DNA complexes in colon adenocarcinoma. Tumor-specific antisera against dehistonized chromatin isolated from transplantable colon adenocarcinoma (from male noninbred Sprague-Dawley rats) were produced. The specificities of these antisera were determined by complement fixation. In the presence of these antisera, only chromatin from colon adenocarcinoma significantly fixed complement, whereas chromatins isolated from normal rat colon epithelia were inactive. Administration of 1,2-dimethylhydrazine to rats produced an early change in the immunospecificity of colon epithelial chromatin similar to that for colon adenocarcinoma. Several lines of experimental evidence indicated that nuclear antigen was not a carcinoembryonic antigen-like substance. Common antigens were also present in human colon adenocarcinomas.


Tumor-associated antigens in cervical cancer tissues and in sera from patients with cervical cancer or with head and neck cancer. Hyperimmune New Zealand White rabbit sera prepared against partially purified tumor-associated antigens (TAA) of invasive cervical cancer tissues (CaCx's) were used to demonstrate TAA in CaCx's and circulating TAA (C-TAA) in sera from patients with cervical cancer or with head and neck cancer. Anti-CaCx serum adsorbed with pooled normal cervical tissue (NCx) antigen preparations and with lyophilized pooled normal human plasma gave precipitin in gel reactions with CaCx but not with NCx, which indicated the presence of TAA in CaCx. The adsorbed antisera reacted with sera from patients with cervical and head and neck cancers, which indicated the presence of C-TAA in such sera. False-positive reactions with control sera from normal humans or from patients with benign gynecologic diseases were much less frequently observed. Statistical analysis of data obtained by the testing of 96 coded serum samples from the National Cancer Institute-Mayo Clinic Serum Bank (Rochester, Minn.) revealed that the results were significant and that specificity was high but sensitivity of the assay was relatively low.


Specific immune response genes of new inbred strains of guinea pigs. Distribution of specific immune-response (Ir) genes controlling responsiveness to synthetic polypeptide antigens, homopolymer of poly-L-lysine (PLL), copolymer of L-glutamic acid and L-alanine (GA) and copolymer of L-glutamic acid and L-tyrosine (GT), and limiting doses of 2,4-dinitrophenyl guinea pig serum albumin  (DNP-GPA) was surveyed in new inbred strains of guinea pigs, JY 1, JY 2, JY 9  and JY 10, established in this Institute. The PLL gene was not found in any of the guinea pigs. The GA gene was found in JY 1 and JY 2 guinea pigs and the GT gene in all the guinea pigs. The gene controlling responsiveness to low doses (1 microgram) of DNP-GPA was found in JY 1, JY 9 and JY 10 guinea pigs. The associated (Ia) antigens was discussed.


Oncofetal antigen: a tumor-associated fetal antigen immunogenic in man. Oncofetal antigen (OFA) has been defined with the use of human natural antibodies as a membrane antigen of human cancer cells that cross-reacts with human fetal brain tissues. The immunogen that elicits the antibody is unknown. The present study was undertaken to examine the immunogenicity of the OFA found on tumor cells. Postoperative melanoma patients were immunized with OFA-positive melanoma cells. Anti-OFA reactivities in the immunized sera were titrated by the immune adherence assay with the use of a known OFA-positive cultured melanoma cell line, M14, as target cell. Alloantibodies were excluded by absorption with lymphoblastoid cells autologous to M14. Anti-OFA antibody then was identified by absorption with fetal brain. In 6 months of immunization, 19 of 23 patients produced increased anti-OFA antibodies. The peak titers ranged from 1:16 to 1:2,048. Sera from 18 patients who were not immunized also were tested for 6 months postoperatively, and none had significant increases in antibody titers. The increase of anti-OFA antibody titer in response to the immunization with OFA-positive tumor cells suggests the immunogenic capability of tumor-related OFA in man.


[Clinico-biochemical principles of healing of myocardial infarct]. Individual glyco- and mucoproteins and collagen metabolites were determined in dynamics in 297 patients with transmural, macrofocal nontransmural, and microfocal myocardial infarction. It is shown that these laboratory findings enable one to appraise the course of myocardial infarction healing. This may serve as one of the criteria for the elaboration of individualized rehabilitation programs.