Two Escherichia coli fructose-6-phosphate kinases. Preparative purification, oligomeric structure and immunological studies. Two isoenzymes of fructose-6-phosphate kinase (ATP: D-fructose-6-phosphate 1-phosphotransferase, EC 2.7.1.11) are present in Escherichia coli K12. One isoenzyme is allosterically inhibited by phosphoenolpyruvate and activated by nucleoside diphosphates, and is a tetramer composed of four subunits of molecular weight 35 000. A simple method for the purification of this enzyme is reported. Equilibrium dialysis indicates that there are four ATP sites and four GDP sites per tetramer. The second isoenzyme is present in low quantity in wild type bacteria. This enzyme is devoid of allosteric properties. A complete method of purification is described. Determination of its molecular weight under native and denaturing conditions indicates that this protein is a dimer composed of two subunits of molecular weight 36 000. Antisera have been produced against both isoenzymes. The antiserum against one isoenzyme does not cross-react with the other. Discrepancies between our results and those of other workers are discussed.


Rat alpha-fetoprotein heterogeneity. Comparative chemical study of the two electrophoretic variants and their Ricinus lectin-binding properties. Two electrophoretic forms of rat alpha-fetoprotein were purified using immunosorbent chromatography and preparative electrophoresis on polyacrylamide gel slabs. Some of their respective chemical properties and their affinity for the Ricinus communis lectin (RCAI) were compared. Electrophoresis on polyacrylamide gradient gel in the presence of sodium dodecyl sulfate indicated a slight difference in molecular 74 000 for the slow alpha-fetoprotein (AFPA) and 72000 for the fat alpha-fetoprotein (AFPB). no significant difference in amino acid composition between AFPA and AFPB was found. A residue of valine was identified at the C-germinal end of both alpha-fetoproteins. The analysis of the CNRr-cleavage products reveals slight differences between AFPS and AFPB. The slow moving alpha-fetoprotein could be further fractionated on RCAI-sepharose column in two components, AFPA1 and AFPA2 differing by their sialic acid content.


Studies on human alpha-fetoprotein. Isolation and characterization of monomeric and polymeric forms and amino-terminal sequence analysis. Human alpha-fetoprotein has been isolated from the serum and ascitic fluid of a patient with hepatoma by a combination of immunoadsorbent column chromatography and Sephadex G-150 gel filtration. Human alpha-fetoprotein is a sialylated glycoprotein with an estimated molecular weight of 67 500, composed of a single-chain polypeptide of approximately 580 amino acid residues and 3.6% carbohydrate. It is a negatively charged protein with an acid isoelectric point (pH 4.57). In addition to the monomeric form of alpha-fetoprotein, we have identified human alpha-fetoprotein polymers, including dimeric and trimeric forms, which dissociate to the monomer only upon exposure to disulfide-reducing reagents, implying that their formation is dependent upon intermolecular disulfide bonds. These polymers are found in human alpha-fetoprotein isolated by isoelectric focusing in both the major (pI 4.57) and minor (pI 5.2) alpha-fetoprotein fractions. The first 17 residues of the NH2-terminal amino acid sequence of the hepatoma-derived human alpha-fetoprotein have been identified. Fetal alpha-fetoprotein is indistinguishable from hepatoma alpha-fetoprotein by several criteria, including immunoelectrophoresis, acryalmide gel electrophoresis, and proclivity for dimerization.


Monosialoganglioside liposome-entrapped enzyme uptake by hepatic cells. Monosialoganglioside liposomes are rapidly taken up by the liver as compared to dicetylphosphate, phosphatidic acid or neutral liposomes. Asialoganglioside GM 1 liposomes are taken up with the same avidity as ganglioside GM 1 liposomes. Competition experiments with asialofetuin suggest that this uptake is mediated by specific recognition of the terminal galactose residues of the glyco-lipid liposomes by the receptor present on the plasma membrane of the parenchymal cells of liver. Thus liposomes containing glycolipids with terminal beta-galactosyl residues should provide an approach for specifically directing biologically active molecules to liver parenchymal cells.


Uptake and degradation of 125I-labelled asialo-fetuin by isolated rat hepatocytes. 125I-Labelled asialo-fetuin was taken up by isolated rat hepatocytes by a saturable process. Half maximum uptake was seen at about 3 - 10(-8) M asialo-fetuin. Non-parenchymal liver cells did not take up asialo-fetuin in vitro. Rate of uptake of asialo-fetuin exceeded rate of degradation at all concentrations of asialo-fetuin tested. Asialo-fetuin consequently accumulated in the cells until the extracellular supply was exhausted. Asialo-fetuin degradation could be studied without concurrent uptake by incubating cells, previously exposed to asialo-fetuin, in asialo-fetuin-free medium. Degradation, as evidenced by increase in acid-soluble radioactivity, was inhibited by NH4Cl and chloroquine. The change with time in the intracellular distribution pattern of radioactivity in cells that had been exposed to 125I-labelled asialo-fetuin for 10 min was examined by means of differential centrifugation. Initially, the radioactivity was found mostly in the microsomal fraction. 60 min after the exposure to labelled protein, the distribution pattern of radioactivity resembled that of the lysosomal enzyme beta-acetylglucosaminidase. The possibility that asialo-fetuin digestion takes place in lysosomes is discussed.


[Substrate specificity and some properties of free and immobilized animal alpha-L-fucosidase]. The effect of a partially purified preparation of pig kidney alpha-L-fucosidase on some glycoproteins--human and rabbit gamma-globulin, glycoprotein from sheep submaxillary gland and ceruloplasmin--was studied. It was shown that the action of the enzyme of the glycoproteins was not accompanied by a release of fucose. A comparative study of the properties of free and concanavalin A-Sepharose 4B-bound alpha-L-fucosidase was done. The experimental data is indicative of difference in the pH-dependenced and thermostability of these two enzyme forms. It was found that bound alpha-L-fucosidase, similar to the free form, did not split off fucose from the native blood group substances. The data of isoelectric fucosing of alpha-L-fucosidase suggests the existence of enzyme polymorphism.


Immunochemical study on serum proteins in systemic sclerosis. Forty one patients with systemic sclerosis were studied after separation into three groups according to Barnett's classification. A multi-dimensional statistical analysis eight serum proteins revealed a difference between control patients and patients with type I and type II scleroderma. Type I scleroderma was characterised by a rise in alpha 2 macroglobulin and in the C4 fraction of complement, whilst in type II scleroderma all the proteins studied were raised, with the exception of CO complement, which was normal, and transferrin which was markedly decreased.


Lymphoid cells of the normal man intestinal mucosa possessing both kappa and lambda light chain specifities. The presence of lymphoid cells possessing both kappa and lambda specificities has been observed in the intestinal mucosa of normal subjects. The variability in the number of such cells in different sections of the same sample and in different subjects seems to be a characteristic of this cell population and may reflect the high activity of the immune system in the gut.


alpha-fetoprotein in rats treated with anti-thymocytic serum. Serum alpha-fetoprotein was detected by immunodiffusion in rats treated with anti-thymocytic serum. Augmented synthesis of AFP is related to compensatory hyperplasia of liver cells secondary to a state of hypoproteinemia in the proteinuric nephritic animals. The nephritis was produced by the anti-basement membrane antibodies present in the thymocytic antiserum.


[Comparative immunochemical analysis of specific beta globulins of the "pregnancy zone" of the humans and other mammals]. By means of the immunodiffusion method it was shown that human pregnancy-specific beta-globulin, as well as analogous beta-globulins in the pregnancy serum of mammal animals (rat, guinea-pig) had an individual antigenic structure. During pregnancy the blood serum of the allied species (rat, mouse) contained immunologically similar beta-globulins which apparently were synthesized in the placenta.


[Macrophage migration inhibition by T-cells in rats after overcoming natural tolerance to alpha fetoprotein]. Cellular immunity to alpha-fetoprein (AFP) was studied in rats by the macrophage migration  inhibition (MMI) test after breakage of tolerance to homologous AFP of rat (RAFP). A single injection of cross-reacting AFP of mouse (MAFP) has induced the capacity of peritoneal exudate cells of rats to react in the MMI test to both MAFP and RAFP. The second injection of MAFP results in reduction of the MMI reaction to both antigens and the appearance of stimulation in the macrophage migration together with further elevation of antibody titer to MAFP and RAFP. The fractionation of rat peritoneal cells has shown the T-cell nature of MMI reaction to both MAFP and RAFP.


[Synthesis and localization of alpha fetoprotein in liver regeneration in mice]. Typical mature hepatocytes constituting not over several per cent of the total amount of preserved hepatocytes served as the principal site of the alpha-fetoprotein (AFP) localization in the liver of mice regenerating after the CCl4 poisoning or partial hepatectomy. Morphologically they failed to differ from the principal mass of hepatocytes and retained an antigen of the bile capillaries on the surface. A change id to the dynamics of the AFP level in the animal serum. Apparently in regeneration of the mouse liver the principal AFP production was realized by mature hepatocytes.


[Dependence of the cytogenetic effect on TEPA concentration in human lymphocyte culture]. The authors studied the cytogenetic action of TEPA (tris/2-methyl-1-azyridinyl) on the human lymphocyte culture. It was shown that the increase of the mutagen concentration from 0.125 to 16.0 microgram/ml the cytogenetic effect for the portion of the aberrant metaphases rose from 6.0 to 61.0%, and for the total number of ruptures - from 7.96 to 116.3. A method of finding the least effective concentration of the substance under study in comparison with control is suggested; for  TEPA it constitutes 0.120 microgram/ml. The percentage of chromatide ruptures remained constant in using different TEPA concentration and constitutes 51.72%. Cell distribution of chromosome ruptures is satisfactorily described by geometrical distribution.


[Sequential staining for G- and C-banding of chromosomes in the analysis of the morphology of the short arms of human acrocentric chromosomes]. Sequential staining for G- and C-banding of acrocentric chromosomes of 8 persons showed that the large heterochromatin region occurred more frequently in chromosome 15 than in chromosomes 13 and 14, and in chromosome 22 more frequently than in chromosome 21. There proved to be no correlation between the size of the heterochromatic region and the short arm of the acrocentric chromosomes. The frequency of occurrence of the satellites in the 8 persons was approximately the same for all the acricentric pairs. The C-banded satellite region of the homologous chromosomes is often heteromorphic.


[DNA content in the Purkinje cells of the rat cerebellum spontaneously infected with Kilham virus]. The DNA content in the cerebellum Purkinje cells of rats naturally contaminated with the Kilham virus and in the animals, in which no infection was observed, was determined cytospectrophotometrically on the Feulgen-stained preparations. The incidence of the cells with hyperdiploid nuclei was shown to be equal in the groups compared. It was concluded that the nonmultiple increase of the DNA content in the Purkinje cells of rat cerebellum was apparently unrelated to the virus infection.


[Highly sensitive electroimmunodiffusion method of detecting antigens on cellulose acetate film]. A highly sensitive electro-immunodiffusion test suggested by the authors for antigen detection on cellulose-acetate films consists of three stages: antigen concentration in a discontinuous buffer system on cellulose-acetate films; antigen detection on the same films by immunodiffusion using standard test system; to detect the precipitation bands the washed films are stained with protein dyes in case the reaction takes place in the zone of vision, or subject to further treatment by means of "plating" the precipitates with antiglobulin antibody or by radioautography. The method permits one to reveal the nanogram levels of alfa-fetoprotein and it may be applied for detection of antigens with different molecular weights and electrophoretic mobility.


[Radioimmunoelectrophoretic determination of alpha-fetoprotein. Standard inhibition curves]. The author suggests an express method of radioimmunological determination of murine and human alpha-fetoprotein with the use of electrophoretic separation of immune-bound and free antigen. The method is comparatively simple, it permits to analyze up to 50 samples in the course of 5 hours. Behaviour of the antibody-antigen system in drawing the standard inhibition curves was analyzed. Inhibition of the antibody-antigen reaction in the zone of low and great concentrations was revealed. The effect described requires further study.


[Properdin and the protein composition of lymph and blood following resuscitation]. It was shown in experiments on dogs that the terminal blood letting with the subsequent reanimation by intrabone autoblood force pumping caused regular redistribution of proteins between blood, lymph, and tissues. Properdine and alpha-globulin retention in interstitium not eliminable by reanimation measures, and also stress secretion of gamma-globulins from the lymph nodes was noted.


[Conduct of Bacillus subtilis transformation in mice under conditions of immunologic suppression of exogenous DNAase 1 activity]. Bacillus subtilis transformation was conducted in the abdominal cavity of mice. The frequency of transformation was considerable decreased when bovine DNA-ase 1 (3-- 5microgram) was injected intraperitoneally to these animals. Immune rabbit gamma-globulins containing antibodies to bovine DNA-ase 1 inhibited in vivo the activity of DNA-ase 1, protected transforming DNA from the hydrolyzing effect of this enzyme. This model can be utilized in search for ways to preserve DNA injected into the animal organism for the purpose of genetical engineering.


[Detection of antigens of endogenous viruses of the C-type during mouse development]. Embryos of the 12th-20th day of gestation, newborn and adult AKR and BALB/c mice were investigated for the presence of mouse C-type virus major structural p30 protein (gs-1) and Gross leukemia virus  type-specific antigen AGLV) by means of radioimmunodiffusion with test systems. The p30 protein was distinctly determined from the 12th day of intrauterine development in both mouse lines; it persisted in the embryo tissues until birth and was detectable also in the AKR and BALB/c mouse tissues from the first days of postnatal development and during the whole life. AGLV was not revealed in BALB/c and AKR embryos and in adult BALB/c mice; however it was found in the AKR newborn mice since the 1st-2nd day after birth. Basing on these data a conclusion was drawn that p30 protein and AGLV were expressed independently according to the radioimmuno-diffusion method sensitivity.


[Histamine-liberating action of MCD-peptide from bee venom]. Histamine release (HR) from the isolated rat mast cells induced by MCD-peptide, mellitin (from the Apis mellifica venom) and compound 48/80 was studied. The dose-response curve, the latent period and temperature dependence of HR induced by MCD-peptide were similar to those of HR induced by compound 48/80. The HR induced by MCD-peptide proved to be an energy-dependent process that was independent of cyclic adenosine-3',5'-monophosphate.


[Dynamics of alpha-fetoprotein concentration in mice of different genotypes during the neonatal period]. A comparative assay of the alpha-fetoprotein (alpha-FP) level in mice of various genotypes (CBA, C3H, C57BL/Se/Sn, BALB/c, CC57W, AKR and nude--nu/nu) was conducted in the course of 3 weeks of postnatal period. The concentration of alpha-FP reached the following levels: the first day 2(-10)-2(-9); the 5th day 2(-8); the 8th day 2(-7); the 15th day 2(-4); on the 22nd day the level was zero. Nude mice which showed the alpha-FP concentration of 2(-2) on the 15th day were an exception. A conclusion was drawn that the alpha-FP synthesis was based not on the athymia of nude mice per se, but upon other unknown factors.


[Characteristics of hepatocytes containing alpha-fetoprotein in regenerating mouse liver]. Induction of the synthesis of the embryo-specific protein--alpha-fetoprotein (alpha-FP) was revealed in the SWR mice of different age after one or several CCl4 treatments. The alpha-FP level elevation was the greatest in mice aged under one month. Morphologically in the liver regenerating after CCl4 poisoning the alpha-FP-containing hepatocytes failed to differ from the principal mass of cells, i.e. in the small cells of young animals and in the large hepatocytes of high ploidy in mice repeatedly poisoned with CCl4. The only feature specific for the alpha-FP-containing hepatocytes in the liver regenerating after CCl4 poisoning was their preponderant localization in the region adjacent to necrosis. Analgous localization of the alpha-FP-containing cells was found in the liver of mice poisoned with two other hepatotoxins--paracetamol and allyl alcohol.


Effects of pesticide seed treatments on Rhizobium japonicum and its symbiotic relationship with soybean. Seventeen Rhizobium japonicum cultures isolated from soybean nodules induced formation of nodules on taproots of soybean plants. All isolates reduced acetylene to ethylene to different extents in vitro. Paper disc assay indicated that two insecticides, lindane (gamma-1,2,3,4,5,6-hexachlorocyclohexane), chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate), and a fungicide, thiram (tetramethylthiuram disulphide) individually or in combination caused significant inhibition of the growth of R. japonicum No. 16. The effects of insecticide-fungicide seed treatments on the nitrogenase activity of soybean plants in nitrogen-fixing capacity, weights of leaves, stems, and nodules were determined. Thiram, singly or in combination with lindane and/or chlorpyrifos, significantly delayed growth of the plants and affected the activity of nitrogenase in the fixation of nitrogen 3 weeks after treatments. No drastic effect of any of the pesticide treatments on soybean plant growth was observed after 8 weeks.


Effects of guanethidine on histamine release during reflex vasodilatation in the dog. 1 The effects of guanethidine pretreatment on the release of [14C]-histamine during the reflex vasodilatation induced in the atropinized gracilis muscle by rapid intravenous administration of noradrenaline, were studied in dogs. 2 After guanethidine treatment the haemodynamic reflex response was completely abolished and no appreciable modification of [14C]-histamine release from the gracilis muscle following intravenous noradrenaline was observed. 3 These results suggest the hypothesis that the withdrawal of the sympathetic discharge represents the mechanism of histamine release during the reflex vasodilatation. Therefore, guanethidine would suppress both the passive and the histaminergic component of the baroreceptor reflex through the abolition of the sympathetic tone.


Pulsion intubation for palliation of carcinoma of the oesophagus. One hundred and eighty-one patients with carcinoma of the upper thoracic oesophagus were intubated perorally using a Procter-Livingstone tube. The mortality was 16-6 per cent but, in the patients who survived, the palliation achieved, as judged by improved swallowing, was considered satisfactory. Factors influencing the success of intubation are also considered.


Axonal transport of [3H]fucosyl glycoproteins in noradrenergic neurons in the rat brain. The transport of [3H]fucosyl glycoproteins has been investigated in the noradrenergic pathway from the locus coeruleus (LC) to the hypothalamus in the rat brain by use of local injection of L-[6-3H]fucose into the LC. Two discrete waves of 3H-glycoprotein pass through the hypothalamus in a caudorostral direction traveling at 4 mm/h (96 mm/day) and 2 mm/h (48 mm/day) respectively. Both waves appear to originate from the LC at approximately 2 h after the injection of [3H]fucose, a time when incorporation into glycoprotein has not yet peaked in the LC. Lesioning the LC-hypothalamic pathway with 6-OHDA, but not carrier solution, blocks both waves of 3H-glycoprotein demonstrating that transport occurs exclusively in the catecholamine axons within the pathway. DEAE-Sephadex chromatography combined with 6-OHDA lesions demonstrates clear differences in the character of 3H-glycoproteins in the two waves undergoing transport. The relative lack of labelling of hypothalamic glycoproteins in the interval before and after these waves suggests that relatively few rapidly transported glycoproteins contribute to the non-terminal axon membrane and are probably primarily transported to the nerve terminal. No evidence for slow axoplasmic transport of [3H]fucosyl glycoproteins is found.


Axonal transport of putrescine, spermidine and spermine in normal and regenerating goldfish optic nerves. Radopactove putrescine, spermidine or spermine was injected into the right eye of normal goldfish and fish in which both optic nerves had been crushed 18 days earlier. Fish were sacrificed 0.25-21 days after injection. Trichloroacetic acid-soluble and -insoluble material was extracted from the right retina and both tecta and assayed for radioactivity (significant differences between left and right tecta suggesting axonal transport). The nature of the radioactivity in the TCA-soluble fraction was determined on an amino acid analyzer. Results indicate that putrescine is not axonally transported in intact goldfish optic nerves, but that during regeneration of the optic nerve large amounts of putrescine are axonally transported at rates similar to the fast component of protein transport. Spermidine appears to be axonally transported both in intact optic nerves and in regenerating optic nerves, and at an intermediate rate of transport; the amount of spermidine transported is significantly increased during regeneration. Spermine is also axonally transported in intact and regenerating nerves, at a rate similar to the rapid rate of protein transport. The amount of spermine transported appears to be slightly less in regenerating than in intact nerves during early stages of regeneration, but increases during later stages of nerve regeneration. The results suggest that putrescine and spermidine may be preferentially transported during nerve regeneration, while spermine and spermidine are transported extensively in intact nerves.


Evidence for direct synaptic interconnections between cat spinal alpha-motoneurons via the recurrent axon collaterals: a morphological study using intracellular injection of horseradish peroxidase. By using intracellular injection of horseradish peroxidase into cat triceps surae alpha-motoneurons we have been able to trace the motor axons as well as the recurrent motor axon collaterals with their synaptic terminals both with the light and electron microscope. In addition to the expected projection to the 'Renshaw cell area', the triceps surae motor axon collaterals were frequently found to terminate within the motor nuclei, where at least some of the terminals made direct synaptic contact with triceps surae alpha-motoneurons.


Role of gangliosides in the uptake and retrograde axonal transport of cholera and tetanus toxin as compared to nerve growth factor and wheat germ agglutinin. Previous investigations have shown that tetanus toxin is transported retrogradely in all peripheral neurons whereas the transport of NGF is confined to adrenergic and sensory neurons. Other macromolecules with molecular weights and general physiochemical properties similar to NGF and tetanus toxin (e.g., cytochrome C, insulin, horseradish peroxidase and bovine serum albumin) are not transported to a detectable extent if injected in comparable molar concentrations. For tetanus toxin, which is transported in all peripheral neurons, it has be assumed that it's retrograde transport depends on properties common to all neurons. In view of the relatively high ganglioside content of the neurons and the high affinity of tetanus toxin for the trisialoganglioside GT1, we studied the influence of gangliosides on the retrograde transport of tetanus toxin as compared to NGF. We included into the study cholera toxin which is known to have a high affinity for the monosialoganglioside GM1 and wheat germ agglutinatinin, a lectin with specific affinity for glycoproteins with N-acetyl-glucosamine residues. Both cholera toxin and wheat germ agglutinin were transported efficiently in all peripheral neurons. Preincubation of 125I-cholera toxin with monosialoganglioside GM1 completely blocked its retrograde axonal transport. The transport of NGF and wheat germ agglutinin was affected neither by various purified gangliosides nor by a mixture of bovine brain gangliosides. The transport of tetanus toxin was only reduced by 50% both by the trisialoganglioside GT1 and the bovine ganglioside mixture.


Retrograde axonal transport of horseradish peroxidase for determining motor projection patterns to the developing limb in Xenopus. Horseradish peroxidase (HRP) injected into developing limb buds of Xenopus laevis tadpoles is carried by retrograde axonal transport to the somata of motoneurones in the ventral horn. Small injection of 10% HRP were found to remain well localised to specified sites in the limb bud. Two types of labelled cells were found: diffusely labelled and granular labelled. Diffusely labelled cells result from axonal damage in the presence of HRP. Granular labelled cells result only from uptake of HRP from the region of the axon endings. No gradular uptake was found from axon shafts. It is concluded that the distribution of granular labelled cells accurately reflects the region of the ventral horn projecting to the site of injection in the limb.


The effect of the retrograde axonal transport of nerve growth factor on the morphology of adrenergic neurones. An injection of nerve growth factor (NGF) into one eye of neonatal rats results in an increase in the tyrosine hydroxylase activity of the ipsilateral superior cervical ganglion. This effect was seen maximally after the intraocular injection of a depot preparation of NGF linked to cellulose. The sympathetic neurones that innervate the eye can be identified by autoradiography after the retrograde axonal transport of either NGF or tetanus toxin labelled with [125I]iodine. It was only those cells having their terminals in the vicinity of the depot preparation. This demonstrates that NGF transported from the periphery to the cell bodies is effectively retained within the transporting cell and is not released to act on extracellular receptors within the ganglion. It is suggested that this specificity of action for NGF reaching the ganglion in this fashion is important during normal development in determining the survival of adrenergic neurones.


[Immunologic properties of various cell fractions of a wild strain and a mutant strain of Saccharomyces cerevisiae Hansen]. Some fractions of the Saccharomyces cerevisiae cell wall have been prepared by the action of Helix pomatia juice on intact cells. Immunosera were obtained by injecting rabbits with these fractions. Immunofluorescence reactions, obtained with these sera, show that some fractions of mannopeptides when extracted from a "smooth-colony" mutant strain, have lost antigenic determinants.


Carcinoma of the esophagus: a new proposal for the evaluation of treatment. An 8-year study of the records of 127 patients with carcinoma of the esophagus revealed that the data necessary to make an evaluation of various methods of palliative treatment frequently were not recorded on the charts. Therefore, it was not possible to recommend one particular paliative treatment for carcinoma of the esophagus. The authors believe that a prospective study is required to evaluate objectively the various modes of treatment for this disease. They propose a grading system with numbered values, so that future studies can be made more objective: in addition such a system would allow comparison of series from different treatment centres.


Serum alpha-fetoprotein levels after hepatic artery ligation and postoperative chemotherapy: correlation with clinical status in patients with hepatocellular carcinoma. Eight patients with hepatocellular carcinoma and positive serum alpha-fetoprotein (AFP) levels were treated by hepatic artery legation and postoperative chemotherapy. Three patterns of clinical response were discerned. First, a marked decrease in AFP levels was found in five patients following the treatment. Three of these patients are alive, all with lowered or normal serum AFP levels at 20, 22, and 60 weeks after operation. Second, two patients displayed only a transient decrease in AFP levels followed by a gradual increase. Third, there was a continuous increase in AFP levels in one patient who showed no clinical improvement. The serum AFP levels in each case appeared to correlate with the prognosis of these patients. Thus, serial measurements of AFP levels may provide an index to assess the clinical result of hepatic artery ligation in patients affected with hepatoma.


Combination chemotherapy of germ cell tumors of the testis with vincristine, adriamycin, cyclophosphamide, actinomycin D and medroxyprogesterone acetate. Forty-three patients with disseminated germ cell cancer were treated with a combination of vincristine, Adriamycin, cyclophosphamide, actinomycin-D, and medroxyprogesterone acetate. All the 43 patients were considered evaluable for response. Thirty-one patients (72%) achieved a complete or partial remission and 14 (32.5%) achieved a complete remission. The patients who attained an objective response obtained a significant prolongation of life compared with the nonresponders (median survival 55 vs. 23 weeks). Responses were seen in all histologic categories and most frequently in patients with metastases confined to the lungs. The major side effects were leukopenia and stomatitis. There were no deaths related to toxicity of the chemotherapy.


Serum alphafetoprotein in hepatocellular carcinoma. The serum alphafetoprotein level (AFP) was studies in 125 histologically verified cases of hepatocellular carcinoma, 66 other malignancies, 74 cases of cirrhosis of the liver, 60 of chronic aggressive hepatitis, 12 of chronic persistent hepatitis, 16 of subacute hepatitis, 36 of acute viral hepatitis, and 13 healthy hepatitis B-surface antigen (HBsAg) carriers. Double immunodiffusion and radioimmunoassay (RIA) were used in all cases. AFP greater than 10 ng-ml appeared in 90% of the cases, and was above 400 ng/ml in 69%. In 80% of those above 400 ng/ml, AFP could also be demonstrated by immunodiffusion. The AFP level in hepatocellular carcinoma was discovered to decline as the age increased. It also appeared to be related to the tumor cell type; the relatively immature cell type was more frequently associated with a higher AFP level. The presence of HBsAg did not influence the AFP level. Although the AFP  in other malignancies and liver diseases ranged abnormally from 14 to 69%, the level did not exceed 400 ng/ml as in our cases of hepatocellular carcinoma (except in one case). Thus, this figure provides a diagnostic serum level of AFP for the identification of hepatocellular carcinoma.


Induction of hepatomas secreting large amounts of alpha-fetoprotein. Hepatomas secreting large amounts of alpha-fetoprotein were induced in Fischer rats by the continuous feeding of 0.06% 3'-methyl-4-dimethylaminoazobenzene. Serum alpha-fetoprotein concentrations in the 59 hepatoma-bearing rats ranged from 92--3260 microgram/ml (mean, 972 microgram/ml; median, 830 microgram/ml) at the time of primary tumor transplantation.


Analysis of the interpolation phenomenon in man evaluated by premature atrial stimulation. After early premature atrial stimulations, returning cycles shorter than the basic cycle were observed in 25 patients. In 4 of them sinus echoes were seen to occur after premature atrial stimulation coupling intervals shorter than those able to induce completely interpolated beats. Such a sequence is unexpected if we suggest a sino-atrial entrance block for explaining the interpolation phenomenon. An alternative explanation can be postulated assuming that all these early premature beats are sinus node re-entries and that the completely interpolated beat is only a sinus re-entry fortuitously occurring at an appropriate interval.


Nucleotide sequence of the 3' terminal third of rabbit alpha-globin messenger RNA: comparison with human alpha-globin messenger RNA. Rabbit and human alpha-globin complementary DNA was synthesized using the primer (dT10)dGdCdC hybridized to globin mRNA with reverse transcriptase, and sequenced using the plus and minus gel sequencing procedure (Sanger and Coulson, 1975; Brownlee and Cartwright, 1977). The complete 3' noncoding region, 89 nucleotides in length, and one third of the coding region of rabbit alpha-globin mRNA have thus been sequenced. These data are compared with the near complete 3' noncoding region sequence of human alpha-globin mRNA obtained in these and previous studies. The two mRNAs are approximately 80% homologous in their 3' noncoding regions, except that the human sequence has an insert of 24 nucleotides. A very similar insert in sequence has been described in human beta-globin mRNA (Proudfoot, 1977).


Cytotoxic effect of a reverse transcriptase inhibitor, AF/ABDP cis, on ovaries of young Xenopus laevis--ultrastructural and autoradiographic study. Observations with the light and electron microscopes showed that a reverse transcriptase inhibitor, AF/ABDP cis which is a rifampicin derivative, had a toxic effect on ovaries of young Xenopus laevis. It induced mitochondrial damage in all observed cells and marked alteration of the ultrastructure of nucleus of the pachytene oocyte. Light autoradiography showed that AF/ABDP cis caused wide variation in nuclear labelling with [3H] thymidine from one pachytene oocyte to another. Somatic cells were more uniformly labelled. These results are discussed in relation to the interpretation experiments upon eucaryote cells in which this drug is used as a reverse transcriptase inhibitor.


[Long-term results of prolonged dicoumarol anticoagulant therapy in 327 peipheral arteriopathic subjects]. 327 patients with peripheral arteriopathy were treated solely with protracted dicoumarol anticoagulant therapy. The indication for such therapy was applied not only in patients in whom the high risk and poor run-off contraindicated revascularisation operations, but also in those in good general conditions at the 2nd or 3rd stage with iliac and femoral thromboses. Two-thirds of the arteriopathics observed were given the anticoagulant treatment for more than two years, with a maximum of eight years. Clinical evolution of the disease is discussed in relation to the treated patients' age and stage, while emphasis is given to the very low incidence of complications and contraindications, despite the long period of treatment.


[Projections of bulbar reticular nuclei and raphe nuclei to lobules VI and VII of cat cerebellar cortex]. Injections of horseradish peroxidase were made in lobules VI and VII of the cerebellar vermis (in the Cat). Serial frontal sections of the brain stem reveal that the lateral and paramedian nuclei were essentially projected onto lobule VI, and the projections from the raphe nuclei onto lobule VII.


The role of gamma-globulin in composition of macroamylase. A macroamylase complex has been artificially generated by combining purified pancreatic and salivary alpha-amylase solutions with physiological amounts of human gamma-globulin. This complex was only partially dissociable at pH 3.5. Salivary alpha-amylase was shown to have higher binding affinity to gamma-globulin than the pancreatic alpha-amylase. Small amounts of macroamylase complex were also formed after the addition of purified salivary or pancreatic alpha-amylase to normal serum. This effect was more distinct when salivary alpha-amylase was added. Electrophoretic examination of serum proteins from subjects with macroamylasemia showed a significant increase of gamma-globulin fraction. Seromucoid of these subjects was also above normal range. Electrophoresis of serum glycoproteins yielded raised values of the fraction moving with gamma-globulin.


Effects of albumin and other proteins during assay of amylase activity. Addition of albumin, gamma-globulin, alpha-casein or submaxillary mucin to the assay system for chromogenic measurement of human or rat amylase with blue starch increased the amylase activity, albumin having the most effect. These proteins seemed to increase the activity by protecting amylase from inactivation. Amylase activity was higher in urine samples showing proteinuria than in urine samples without detectable protein. It is concluded that amylase assay is more reliable when a final concentration of albumin of 1 mg per ml is added at the dilution step and at the incubation step.


Electroradioimmunoassay: a sensitive method for the quantitation of alpha-fetoprotein. A sensitive technique, electroradioimmunoassay has been adapted for the estimation of alpha1-fetoprotein in normal adult human plasma and serum. Commercially available antisera were used and a 10-fold increase in sensitivity was obtained in comparison with two conventional radioimmunoassays. Plasma alpha 1-fetoprotein levels of 2.6 +/- 1.2 microgram/1 (mean and standard deviation) were found in a normal population as compared with levels of 4.1 +/- 2.6 microgram/l in a group of 49 patients with alcoholic liver disease.


Cord serum levels of IgM, IgA, and alpha-fetoprotein and their interrelationships. IgM, IgA, and AFP concentrations were determined in a group of 30 umbilical cord sera chosen at random from the cord sera sampled over a period of one year. Because low levels were expected, use was made of sensitive techniques, which were standardized on the basis of international reference preparations. Geometric mean levels were 5.58 mg/l for IgA, 142 mg/l for IgM and 53.6 mg/l for AFP. Statistical analysis of the distinct IgM, IgA, and AFP levels showed no significant correlation with either birth weight or birth length. The difference between the sexes in the levels of these proteins was not significant either. The difference between the IgA concentration in sera from children of primipara and those of multipara mothers was significant. Moreover, a statistically significant negative correlation was found between the IgA and AFP levels. The results are discussed in relation to current knowledge about the ontogeny of humoral immunity in man and a possible biological function of AFP in ontogeny.


Methods for semi micro or automated determination of thrombin, antithrombin, and heparin cofactor using he substrate, H-D-Phe-Pip-Arg-p-nitroanilide-2HCl. Methods for the measurement of thrombin and plasma antithrombin, by end point determination at a semi micro level and also by rate assay measurement in a fully automated system have been devised using the thrombin specific chromogenic substrate, H-D-Phe-Pip-Arg-p-nitroanilide. Preliminary defibrination of plasma is avoided in both methods. The semi micro method has been correlated with antitrhombin measured in plasma of postoperative patients by established clotting and immunological assays. The automated method has been found to be highly reproducible and to have less scatter than the other procedures.


Serum FSH, LH and prolactin in normal males and patients with prostatic diseases. Serum FSH, LH and prolactin were measured in fifty-eight normal males between 30 and 80 years of age. At the same time similar estimations were performed on samples taken from 232 patients with benign prostatic hypertrophy (BPH) and twenty-six patients with prostatic carcinoma. The three groups were compared with respect to age, and it was observed that significant rises related to age occurred in the serum levels of FSH, LH and prolactin in normal men after the sixth decade. The patient groups did not differ significantly from each other, or from the normal age-matched population in respect to prolactin and FSH levels. Serum LH in both the carcinoma and BPH patient groups, however, differed significantly from the controls, and remained at the level associated with younger normal males. It is suggested that testosterone metabolites from the prostate exert a negative feedback on pituitary LH secretion.


Determination of cryoglobulins as lipoprotein-autoantibody immune complexes and antigenic determinants against antilipoprotein autoantibody. Serum IgG-antilipoprotein-autoantibody activity (at 4 degrees C) of a plane xanthoma patient was shown by double-immunodiffusion method. Cryoglobulins in the serum were dissociated to polyclonal IgG and alpha- and beta-lipoproteins by acidification and were reconstructed by neutralization. IgG fraction of the cryoglobulins precipitated with lipoproteins. The cryoglobulins were thus demonstrated to be immune complexes of polyclonal IgG-antilipoprotein-autoantibody and both alpha- and beta-lipoproteins. A part of the lipoprotein-autoantibody immune complexes was not cryoprecipitable. Antigenic determinants for the autoantibody existed in the lipid moieties of lipoproteins, in contrast to the apoproteins which determined the specificity to heteroimmune antilipoprotein antibody. The presence of more than nine different antigenic determinants against the autoantibody indicated that lipoproteins were immunologically heterogeneous depending upon the lipid moieties. Lipoproteins reactive with the autoantibody varied quantitatively in normal individuals and were not detected in a primary hyper-beta-lipoproteinaemia patient and in a primary biliary liver cirrhosis patient with much lipoprotein-X. The absence of antigenicity in the two patients' sera is most likely caused by abnormal lipid moieties of lipoproteins.


Identification of pregnancy-associated alpha-macroglobulin on the surface of peripheral blood leucocyte populations. Pregnancy-associated alpha-macroglobulin (PAM) was identified on the surface membranes of human peripheral blood leucocytes by immunofluorescent staining. Subjects having a high serum-level of the glycoprotein (pregnant and contraceptive steroid-treated women) were shown to possess a significantly greater proportion of PAM-positive cells than those (normal males) with a low concentration. However, no correlation was found between the serum values and the percentage of PAM-associated leucocytes. Studies employing purified leucocyte-types and the simultaneous detection of PAM and lymphocyte surface markers indicated that only subpopulations of T lymphocytes (23%) and monocytes (40%) were associated with this serum protein.


Antibody-dependent cellular cytotoxicity (ADCC) against Epstein-Barr virus-determined antigens. III. Reactivity in sera from patients with Burkitt's lymphoma in relation to tumour development. Sera from patients with Burkitt's lymphoma (BL) were tested for antibody-dependent cellular cytotoxicity (ADCC) against Epstein-Barr virus (EBV) determined antigens and for antibodies directed against various EBV-specific antigens. No correlation was found between EBV serology and ADCC in sera selected for high or low titres against the membrane antigens (MA) or in consecutive sera from three patients with isolated late tumour recurrences. Furthermore, no correlation was found between ADCC and tumour development.


Potentiation of experimental allergic thyroiditis in rats adipectomized at birth. The effect of neonatal extirpation of the interscapular brown adipose tissue (adipectomy) on the development of experimental allergic thyroiditis in 8-week-old inbred Lewis rats was studied. The incidence and intensity of disease were much higher, and delayed skin reactions significantly stronger, in adipectomized rats than in control sham-adipectomized rats.


Cytomegalovirus-directed lymphocyte reactivity in healthy adults tested by a CMV-induced lymphocyte transformation test. A cytomegalovirus (CMV) induced lymphocyte tranformation test (LTT) performed as a microtechnique assay is described. Investigations were done in a group of twenty-five healthy adults with normal lymphocyte reactivity to phytohaemagglutinin and previously encountered antigens. Lymphocyte reactivity was established in CMV-seropositive persons using heat-inactivated cell-free CMV strain AD 169 in an optimal stimulating dose of 10(1-5) TCID 50. Positive CMV-LTT results were shown to be induced by purified inactivated CMV. Most persons (twelve out of seventeen) with antibodies against CMV-induced late or virus-structure antigens (LA) showed positive (stimulation index greater than 2-0) CMV-LTT results. Magnitude of CMV-LTT responses was positively related to the CMV-LA Ab titres (P less than 0-01). All persons (six) with antibodies against CMV early or non-structural antigens showed positive CMV-LTT results and highest stimulation rates were found in this group. Generally CMV-seronegative persons were non-responsive. Only in one case (LA Ab titre 10) a borderline positive response (stimulation index 2-3) was seen. The CMV-LTT was shown to have virus-specific properties because it was not related to simultaneously performed LTTs with Epstein-Barr virus and Herpes simplex virus in the same persons. It is concluded that CMV-LTTs are useful virus-specific in vitro techniques for the study of CMV-directed lymphocyte reactivity. Its possible relationship to other cell-mediated immunity tests and its practical applications are discussed.


Monoclonal IgG3-kappa and IgG1-lambda proteins with different idiotypic determinants present in a single patient. Two monoclonal proteins belonging to IgG3-kappa and IgG1-lambda subclasses were detected in a single patient. Immunochemical analysis revealed that the idiotypic determinants of the two proteins were different. Examination of bone marrow by an immunofluorescence technique indicated that these two proteins were produced in different cells, although both kappa and lambda chains were present in a minute proportion of the cells. J chain was present in all cells that produced IgG.


Immune responses of BALB/c mice to the idiotype of T15 and of other myeloma proteins of BALB/c origin: implications for an immune network and antibody multispecificity. Normal adult BALB/c mice are virtually unresponsive to the idiotype of myeloma protein T15 when immunized with purified T15. However, antibodies to the T15 idiotype can be elicited by T15 in BALB/c mice that are reared in a germ-free environment or injected as newborns with an alloantiserum to the idiotype. Some germ-free and neonatally suppressed mice also produce helper cells (presumably T helpers) that enhance B-cell production of anti-DNP antibodies in response to DNP-T15. Taken together with previous studies, the present results mean that so far there are no exceptions to the rule that BALB/c mice have B and T cells that can respond to idiotypes of myeloma proteins of BALB/c origin. There appears to be a reciprocal relation between the natural prevalence of an immunoglobulin's idiotype and its immunogenicity in isologous individuals. The findings support proposals for an immune network of idiotypes and anti-idiotypes. Besides binding one or more extrinsic antigens, it is likely that each immunoglobulin also binds the idiotype of some other immunoglobulin. An immune network therefore implies multispecificity of antibodies.


Has small-scale audio-visual dental health education a viable future? The achievement of an adequate oral hygiene status in large groups of people remains an unfulfilled objective of the dental profession. In this study an attempt was made to motivate 180 pregnant women by showing them a tape-slide program. The subjects were randomly allocated into four statistically equal experimental groups. Two examiners, who were calibrated before commencement of the survey, measured Gingival and Plaque Indices of each subject and a questionnaire was used to assess the educational potential of the tape-slide sequence over a period of 4 weeks. There was a 100% incidence of gingivitis in the study population. Those women who viewed the tape-slide program in addition to being dentally examined, achieved the greatest reduction in their GI and retained the most information. The tape-slide sequence alone did not initiate a satisfactory degree of behavioral change; similar studies have also had discouraging results and it would appear that individual attention and motivation is the only sure way to date of achieving adequate personal plaque control.


Contact urticarial skin responses to cinnamaldehyde. A patient suffering from an eczema of the right hand showed after routine patch testing an immediate urticarial reaction to cinnamaldehyde. The same reaction could be noted in three out of eight control persons. Laboratory investigations revealed cinnamaldehyde to have a histamine liberating effect, a fact not previously mentioned in reports on untoward reactions to this compound.


The antigenicity of soluble porcine elastins: I. Measurement of antibody by a radioimmunoassay. Immunization of rabbits with either porcine tropoelastin, lung alpha-elastin, or aortic alpha-elastin resulted in the production of antibodies against the respective antigens. The assay of antibody activity with a radioimmunoassay indicated a high degree of cross reactivity between these three soluble elastin preparations. Through the use of competitive protein binding experiments it was possible to detect antigen-specific differences between tropoelastin and the two alpha-elastins. No antigenic differences were observed between lung or aortic alpha-elastin by any of the assay procedures uded in this investigation. Absorption of the various antisera with either insolubilized tropoelastin, lung elastin or aortic elastin allowed the preparation of non-crossreactive antibodies and provided further evidence for the antigenic differences between tropoelastin and the soluble alpha-elastins. The implications of the findings, as they relate to our current understanding of the molecular and structural arrangement of elastin and the elastin precursor, are discussed.


Hoechst 33258 banding of Drosophila nasutoides metaphase chromosomes. Hoechst 33258 banding of D. nasutoides metaphase chromosomes is described and compared with Q and C bands. The C band positive regions of the euchromatic autosomes, the X and the Y fluoresce brightly, as is typical of Drosophila and other species. The fluorescence pattern of the large heterochromatic chromosome is atypical, however. Contrary to the observations on other species, the C negative bands of the large heterochromatic chromosome are brightly fluorescent with both Hoechst 33258 and quinacrine. Based on differences in the various banding patterns, four classes of heterochromatin are described in the large heterochromatic chromosome and it is suggested that each class may correspond to an AT-rich DNA satellite.


Visualization of ribosomal gene activity: silver stains proteins associated with rRNA transcribed from oocyte chromosomes. Cricket oocyte chromosomes were stained with silver at pachytene when certain chromosome regions are active in rDNA amplification and rRNA transcription. The silver preferentially stained the known locations of 18S + 28S ribosomal cistrons. Cytochemical tests revealed that the silver binds neither to the rDNA nor transcribed rRNA, but rather to proteins which rapidly associate with the freshly-transcribed rRNA. As rRNA transcription proceeds, the quantity of silver stainable proteins progressively increases. The silver procedure can be used to visualize gene activity at the rDNA sites with conventional light microscopy.


[Pre-stained slides for differential blood counts (author's transl)]. A new method for differential blood counts using pre-stained slides showed it to be suitable for routine use. To differentiate white blood cells the same experience was needed as for conventional stainings. Normal and abnormal blood pictures can be reliably distinguished. The particular advantage of the method is in the simple preparation of the smear and in avoiding the need for time-consuming staining.


Autoradiographic and tissue distribution studies on a nitrovinylfuran derivative (SQ 18506 14C) in S. mansoni infected mice. Autoradiographic and tissue distribution studies of SQ 18506 14C were carried out on 13 Swiss albino mice. Infection was done by I.P. route with 50 to 60 cercaria of S. mansoni (puerto Rican strain) per animal. The safe single i.v. dose of a solution containing one mg of SQ 18506 dissolved in 0.06 ml dimethylsulfoxide was 0.003 ml/g b.w. of mice. Schistosome autoradiograms were clearly demonstrated one day after that dose injected in each mouse 50 days post-infection. However, hepatic autoradiograms were visible 14 days after 2 doses of our drug to each infected mouse. The higher the concentration of SQ 18506 14C in the culture medium the darker were the schistosomes in the autoradiograms and the greater their total d.p.m/mg. Tissue distribution studies after 2 1/2 doses of our drug/mouse revealed that schistosome total d.p.m/mg dry-weight was 50 times more than that of its liver. The latter d.p.m/g wet-weight was slightly higher than that of one ml of mouse blood.


Biotransformation of morphine to dihydromorphinone and normorphine in the mouse, rat, rabbit, guinea pig, cat, dog, and monkey. Biotransformation of morphine to dihydromorphinone and normorphine was studied in several mammalian species. Free and total dihydromophinone, morphine, and normorphine in the urine were determined, as propionyl derivatives, with a gas-chromatographic technique. Dihydromorphinone was detected as a morphine metabolite in the acid-hydrolyzed urine of all species studied except the dog and morphine-dependent man. Normorphine in both free and conjugated forms was detected in the urine of all species studied. The degree of biotransformation of morphine to dihydromorphinone in the guinea pig did not change during chronic administration of morphine sulfate, 25 mg/kg, daily for 28 days. The small amounts of dihydromorphinone and normorphine produced as metabolites make it unlikely that they play any significant role in the modification of the pharmacologic effects of morphine.


Immunogenic contaminants in mouse nerve-growth factor. Experiments are described that confirm the presence of gamma-globulin in standard preparations of mouse nerve growth factor (7-S complex and beta subunit) and show that antibodies to this protein are present in horse antisera to the growth factor. These antibodies may be partially removed by adsorption with soluble antigen and totally removed by affinity chromatography on columns of insolubilised antigen. These procedures do not affect the potency of the antiserum in vitro or in vivo. Contaminating globulin may be removed from samples of the isolated beta subunit by gel filtration to give an immunochemically pure preparation which will permit more meaningful studies on nerve growth factor and its antiserum.


Single-drug chemotherapy of bladder cancer with adriamycin, VM-26 or bleomycin. A phase II multicentric, co-operative study. Single-drug chemotherapy was employed in a multicentric co-operative trial. Adriamycin was given to 18 patients, VM-26 to 30 and Bleomycin to 33. Almost all patients were suffering from a T3 or T4 bladder cancer with known metastases in almost 40% of cases. The vast majority of patients had already received previous treatment either chemotherapy, surgery of irradiation. Complete regression was obtained in 5% of cases; partial objective regression in 11.1% of patients treated with Adriamycin, 26.6% with VM-26 and 33.3% with Bleomycin. The results were uniformly better when high dosages of drug were employed. No clear-cut superiority of one drug over the others can be demonstrated.


Sequential analysis of supraventricular arrhythmias and its value in the differentiation of irregular rhythms. Distinctive electrocardiographic patterns for atrial fibrillation, sinus rhythm, and atrial extrasystoles and sinus arrhythmia were found by analysing the sequential distribution of cycle lengths. Sequential analysis demonstrated similarities between those arrhythmias, accounting for confusion between them, but it could also be used to demonstrate important differences. Utilizing information from both the frequency distribution and the sequential distribution of cycle lengths has improved the accuracy of computer identification of 20-sec arrhythmia recordings from 58.4 to 70.5%, when all supraventricular arrhythmias recorded on acute medical admissions were considered. It is suggested that such information will improve arrhythmia identification based on P wave analysis. From this analysis a classification of irregular rhythms has been proposed. An irregular rhythm may or may not be totally irregular. If there is lack of sequential dependency an arrhythmia should be described as nonperiodic. If there are distinct cycle lengths which are most frequent, the rhythm may be described as modal and there may thus be bimodal or trimodal rhythms which are irregular. Sinus rhythm with atrial extrasystoles may show a combination of these features. This can be described as a bimodal nonperiodic rhythm. If there is neither sequential dependency nor distinct modality the rhythm may be described as an irregular nonperiodic rhythm, e.g. some examples of atrial fibrillation. Sinus arrhythmia is an irregular periodic rhythm.


Macroamylasaemia after treatment with hydroxyethyl starch. After infusion of 500 ml of 6% hydroxyethyl starch into fifty-four patients an increase of serum amylase was observed which in fifty-one cases exceeded the upper limit of normal (190 U/l). In most cases serum amylase reached twice the basal value. Renal function influenced the duration of the increase in serum amylase, but not the maximum increase (201+/-15 U/l; mean+/-SEM). In patients with advanced renal failure (glomerular filtration rate (GFR) = 2-10 ml/min) serum amylase was still markedly elevated after 72 h (298+/-24 U/l; mean+/-SEM). In patients with normal renal function (GFR greater than 90 ml/min) serum amylase decreased to 183+/-40 U/l (mean+/-SEM) within 72 h without reaching basal values. After infusion of HES no changes were observed in serum lipase or in amylase or lipase activities in duodenal secretion. Amylase excretion in the urine decreased. The assumption of a macroamylasaemia caused by formation of an HES-amylase complex was confirmed by gel filtration. The elimination from plasma of this high molecular enzyme-substrate complex is slow and causes hyperamylasaemia. In no case was the macroamylasaemia associated with signs or symptoms. An awareness of this causal relationship seems to be important, to avoid the erroneous diagnosis of a pancreatic disease.


In vitro secondary mixed leukocyte reaction (MLR). II. Interaction MLR determinants expressed by F1 cells. T cells from strain A primed in vitro to (C57BL/6 x A/JF1 [(B6 x A)F1] cells respond better to restimulation by (B6 x A)F1 than by B6 or a 1:1 mixture of A and B6 cells. The increase in the response to F1 cells is specific and due to MLR determinants present on (B6 x A)F1 cells but not on either of the parental cell types. (B6 x A)F1 cells express more than one F1-specific MLR determinant, and this expression is dependent upon products of alleles of at least two loci within the major histocompatibility complex (MHC). Responsiveness to these F1-MLR determinants is apparently controlled by more than one locus within the MHC.


Transfer of idiotype suppression and idiotypes by leukocytes from ascitic fluids. A procedure is described for producing ascites containing both high concentrations of antibodies and immunologically active leukocytes. In mice that were immunologically suppressed with respect to a cross-reactive idiotype, the ascitic cells were found to be very active in adoptively transferring the suppressed state to mildly irradiated, syngeneic recipients and simultaneously transferring the capacity to produce the "private" idiotype characteristic of the donor's anti-p-azophenylarsonate antibodies. The participation of both T and B cells is therefore suggested. As few as 2 x 10(6) cells were sufficient for suppression and for inducing the production of the private idiotype in the recipient, in concentrations comparable to that of the donor mouse. A single donor can thus be used to colonize many recipients with the private idiotype, without sacrifice of the donor. The method should be useful in structural studies of antibodies of a given specificity with different idiotypes, and for investigations of the mechanism of idiotype suppression, as well as other studies in cellular immunology.


In vitro studies on H-2-linked unresponsiveness to synthetic polypeptides. III. Production of an antigen-specific T helper cell factor to (T,G)-A--L. The in vitro production, assay and partial characterization of an extremely potent antigen-specific T helper cell factor to poly-L-(Tyr, Glu)-poly-DL-Ala--poly-L-Lys [(T,G)-A--L] which may be an important physiological mediator of T cell function in vivo is described. The factor has an antigen-binding site, Ia and Ig-like determinants on the same molecular complex. The response to (T, G)-A--L is under H-2-linked Ir gene control, some but not all low responder haplotypes will produce the factor.


Ia-bearing cells promote the concanavalin A mitogenic response of Ia-negative T cells. Nylon wool column-purified T cell populations depleted of Ia-bearing cells do not respond to the T cell mitogen concanavalin A (Con A). The response of this population can be restored by admixture of untreated nylon wool-nonadherent T cells. These promotor cells express Thy-1, are not sensitive to mitomycin C treatment and cannot be replaced with adherent spleen cells. Experiments using the T6 chromosome marker demonstrate that Ia-depleted populations respond to ConA only in the presence of the Ia-positive population.


Detection of bursa and thymus-specific alloantigens in the chicken. CH strain chickens selected from the F2 progeny of a cross between CH (B10/10) and B14B (B14/14) strains carried bursa (BA) and thymus (TA)-specific alloantigens of B14B parental origin. The BA marker was present on 95% of bursal cells but only on 10% of splenic and blood B lymphocytes. The TA marker was expressed by 80% of thymus cells and weakly by 45% of splenic and 70% of blood T lymphocytes. The adopted immunological and genetic protocol offers a feasible approach towards detection of differentiation antigens in major histocompatibility complex-homozygous but 'minor' alloantigen-segregating populations.


Failure of T cells specific for strong histocompatibility antigens to cooperate with B cells for a humoral response. Thymus-derived lymphocytes (T cells) selected for reactivity to strong histocompatibility antigens over a period of one to twelve months in vitro were tested for their ability to cooperate  with bone marrow-derived lymphocytes (B cells) for a humoral response. If cultured with normal syngeneic or allogeneic spleen cells and sheep erythrocytes (SRBC) as immunogen, inhibition of the anti-SRBC response was observed. Similarly, in T cell-free spleen cells the alloreactive T cells did not stimulate a humoral response, indicating that they cannot exert cooperative activity on B cells. Since it was possible that the alloreactive T cells may cooperate with B cells in a humoral response to immunogens carrying histocompatibility antigens, hapten-coupled tumor cells were also used as immunogen. Though it is demonstrated that the alloreactive T cells do recognize the tumor cell immunogen, no stimulation of the B cells for a humoral response against the tumor cell is observed. This result, as well as the finding that the antigenic requirements for T helper cell priming and cell proliferation of the alloreactive T cells are different, suggests that these alloreactive T cells and helper T cells belong to different T cell subsets.


Restriction in the immune response to flagellar proteins in inbred mice. Anti-idiotypic sera prepared in both AL/N mice and rabbits identify specificities common to the IgA MOPC 467 myeloma protein and day-7 affinity-absorbed antibodies to Salmonella milwaukee polymerized flagellin (S. mil-POL) raised in BALB/c and C57BL/Ka mice. Isoelectric focusing (IEF) of reduced and alkylated BALB/c anti-S. mil-POL gave a banding pattern of L and H-chains identical to MOPC 467. C57BL/Ka anti-S. mil-POL had a similar L-chain pattern but a different H-chain pattern. Both BALB/c and C57BL/Ka contain predominantly IgA. The IEF patterns in the two strains are consistent with a monoclonal response at day 7.


The nature of prostaglandin-like substances released from guinea-pig lung in anaphylaxis. The pulmonary inactivation of infused prostaglandin E2 by isolated perfused lungs of guinea pigs was inhibited with di-4 phloretin phosphate (DPP) without significantly altering the antigen-induced release of prostaglandin-like substances or histamine. Similar data were obtained when the lungs were perfused in a reverse direction. These results suggest that parent prostaglandins are not major constituents of the prostaglandin-like substances released from guinea-pig lungs during anaphylaxis.


Allergic reactions, cyclic AMP and histamine release. Both isobutylmethylxanthine and theophylline increased the level cyclic AMP in the mast cell. Theophylline reduced the allergic histamine release, whereas isobutylmethylxynthine caused a pronounced potentiation of the histamine release. This indicates that the hypothesis of an inverse relationship between the level of cyclic AMP in mast cells and histamine release is too simple.


Radiation-released histamine in the rhesus monkey as modified by mast-cell depletion and antihistamine. 4000 rads of mixed gamma neutron radiation administered to rhesus monkeys released a significant amount of histamine into their circulation. When the monkeys were treated with a mast-cell histamine depleter (compound 48/80) for 4 days and then irradiated, no increase in circulating histamine was seen. When 48/80 was give 20 min after irradiation, only a slight increase in histamine was seen, indicating that 4000 rads had released most of the mast-cell histamine.


An electrophoretic investigation of the binding of 3-14C coumarin to rat serum proteins. The binding of coumarin to serum proteins of the rat has been demonstrated. Of the total bound coumarin (37% of injected dose), 36% was bound to slow and fast oc1 globulins, 11% to the post albumins, 10% to globulin and 9% to albumin.


HLA antigens in diabetic children.

HLA antigens in diabetic children. Ninety insulin-dependent diabetic children were HLA-typed in order to elucidate the role played by HLA complex-linked genes in the pathogenesis of diabetes mellitus of childhood. HLA-Aw30 and HLA-Bw35 were significantly increased and decreased, respectively, in the diabetic group as compared with controls. In relation to age at onset of diabetes, HLA-B8 was significantly increased in the 0-5-year group. By dividing the patients according to the season at onset of the disease, only HLA-Aw30 in the October-January group reached the level of significance.


Early lesions and development of primary hepatocellular carcinoma in man--association with hepatitis B viral infection. It is well known that primary hepatocellular carcinoma could be derived from chronic hepatitis and liver cirrhosis in epidemiologic studies. However, it is still not clear what kinds of hepatocyte are premalignant cells. Recently we have focused on liver cell dysplasia as a possible premalignant cell, and showed localization of alpha-fetoprotein in the cytoplasma of these cells. Although the dysplastic cells were often seen in the liver of chronic active hepatitis, hepatitis B virus associated DNA polymerase activity was also significantly high in the sera from the patients with chronic active hepatitis. In this paper, we discuss the possible role of hepatitis B virus through hepatocarcinogenesis in human.


Argentaffin and argyrophil reactions of human gastrointestinal carcinoids. Forty-five carcinoids from the gastrointestinal tract, classified on morphological grounds, were stained with an argentaffin technique (Masson-Hamperl, Virchows Arch [Pathol Anat] 266:509-548, 1927) and three argyrophil techniques (Grimelius, Acta Soc Med Upsal 73:243-276, 1968; Sevier-Munger, J Neuropathol Exp Neurol 24: 130-135, 1965; and Hellerström-Hellman, Acta Endocrinol (Kbh) 35:518-532, 1960). Most but not all carcinoids from the embryonal midgut were argentaffin, whereas most fore- and hindgut carcinoids were nonargentaffin. The argyrophil stain of Sevier-Munger stained all argentaffin tumors and in addition a few nonargentaffin foregut carcinoids. The Grimelius stain stained all and the Hellerström-Hellman stained none of the tumors, irrespective of their location. In the argentaffin carcinoids the frequency of argyrophil cells exceeded that of argentaffin cells.


Gastrin cell immunofluorescence in conventionally fixed and stained tissue sections. We describe a method for visualizing gastrin cells by immunofluorescence that can be applied to tissues conventionally fixed and previously stained with hematoxylin and eosin. It results in a clear demonstration of surrounding structures. This technique permits the use of a broad of histological specimens for gastrin immunofluorescence and may also prove useful in the immunohistochemistry of other gastrointestinal endocrine cells.


Distribution of nuclei in macroconidia of Microsporum gypseum. The distribution of nuclei in macroconidia was investigated in 20 strains of the dermatophyte Microsporum gypseum by fluorescence microscopy. The macroconidia were characterized by the right-left and the central-marginal symmetry indices. In 18 strains the significant majority of the nuclei were situated in the basal part of the macroconidium, the remaining 2 strains showed a more or less uniform distribution of nuclei. All strains had higher numbers of nuclei in the central part of the macroconidium as compared with the periphery.


Exclusion of the HLA locus from a large portion of the long arm of chromosome 6. HLA antigens were determined in two infants with multiple congenital anomalies and in their healthy parents and one sibling. One infant had a deletion of a major portion of the long arm of chromosome 6. The other child had a translocation of a similar piece of chromosome 6 to the short arm of chromosome 3. The mother and the maternal grandmother showed this translocation in a balanced state. The HLA types of both children and their parents exclude the localization of the major histocompatibility locus from the deleted or translocated portion of the long arm of chromosome 6.


Cellular requirements for the induction of cytotoxic T cells in vitro. The stimulator and accessory cell requirements for the induction of cytotoxic T cell responses to alloantigens in vitro are reviewed. The ability of lymphocytes to stimulate was acquired early in development and was found to be a property of both Ig+ and Ig- cells. The presence of Fc receptors, complement receptors or Ia antigens on lymphocytes did not confer superior stimulator capacity. In contrast to lymphocytes mature macrophages were poor stimulators. When resting lymphocytes were used as a source of stimulator cells there was an additional requirement for an adherent accessory cell population for the induction of cytotoxic T cells. The accessory cell population functioned whether syngeneic or allogeneic with the responder and was required for the induction rather than the maintenance of the response. It was further characterized as an Ig+, theta, FcR+/-, CR +/-, Ia- cell which was not a mature macrophage. Accessory cells could be replaced to some extent by supernatants from spleen cell cultures. A model for T cell induction is proposed where only cells which produce unique inductive stimuli are able to function as stimulator cells. Stimulator cells are defined as those cells capable of stimulating cytotoxic T cell precursors directly and independently. Stimulator capacity is suggested to be a function of activated cells only, thus lymphocytes only stimulate following activation by accessory cells. Cells lacking innate stimulator capacity but exhibiting Ia antigens are suggested to stimulate via an alternative means.


Responsiveness to HY antigen Ir gene complementation and target cell specificity. The H-2 restricted nature of the cytotoxic T cell responses to the HY antigen in mice has been reviewed, together with mapping data for the H-2 K and/or D association of responses to male cells of the following haplotypes: H-2b, H-2d, H-2k, H-2s. Ir gene complementation is implicated in the HY response of a number of F1 mice derived from non-responder parental haplotypes, and in one recombinant strain. Thus, at least two and possibly more Ir genes are involved in the anti-HY response, in addition to the K and/or D gene products with which HY is obligatorily associated, certainly at the target cell level.


[Hyperamylasemia following infusion of hydroxyethyl starch with different molecular weight distributions]. The infusion of hydroxyethyl-starch leads to hyperamylasemia due to the complex structural interrelation of the amylase molecules in the serum and the hydroxyethyl-starch molecules. The amylase clearance is reduced because glomerular filtration of the macromolecules thus formed is not possible. The extent of hyperamylasemia is dependent on the intravascular remission duration of the hydroxyethyl-starch preparation used. The question, whether and to what extent an enzyme induction in the liver also contributes to the observed hyperamylasemia, remains open.


Deposition in the lung and uptake to blood of motor exhaust labelled with 203Pb. Inhalation of airborne lead from automobile exhaust is a significant source of intake. In these experiments a radioactive isotope of lead was used to label tetraethyl lead consumed in petrol engines. The resulting aerosols were inhaled by human volunteers to determine the percentage deposition in the lung, the pattern of removal from the lung and the resulting levels in the blood and excreta. Deposition in lung was shown to depend on particle size, varying from 40% for fresh, highly diluted aerosols to 14% for mature, aggregated aerosols, at a breathing rate of 15 breaths per minute. For aggregated aerosols the influence of rate of breathing, tidal volume and reserve volume was investigated. Uptake from the lung was rapid (T 1/2=6.6 h) and virtually complete with little, if any, ciliary clearance. Fifty per cent of the labelled lead remained in the blood, the concentration peaking after 2 days and subsequently declining exponentially (T 1/2 = 16 d). Total excretion was of the order of 1% of the intake per day up to 15 days.


Adjuvanticity (immunity-inducing property) of cord factor in mice and rats. Cord factor, a glycolipid in mycobacteria, was found to make a tolerogenic protein antigen immunogenic when injected 0 to 2 days before tolerogen injection in mice and rats. Cord factor was also found to increase phagocytic function of the reticuloendothelial system of mice, and the relationship between the immunity-inducing capacity and the phagocytic function is discussed.


Immunological analysis of plasma membranes of a T-strain of mycoplasma (Ureaplasma urealyticum). The cell membranes of a T-strain of mycoplasma, obtained by ultrasonic disruption, were as effective as whole organisms in eliciting metabolism-inhibiting and complement-fixing antibodies. The soluble fraction separated from cell membranes by centrifugation at 35,000 X g showed a minor ability to elicit an antibody response as measured by metabolism inhibition and complement fixation tests. After a further centrifugation at 100,000 X g, the immunogenic activity of the soluble fraction was completely lost. Immunogenic determinants in mycoplasma membranes could also be demonstrated by adsorption tests; cell membranes were more effective than soluble fractions in adsorbing antibody capacity from the immune sera against whole cells. It has been shown by further experiments that cell membranes have at least two major antigenic determinants, which differ either in chemical nature or in capacity to adsorb and evoke antibodies, characterized by different serological behaviors.


Synthesis and cleavage processing of oncornavirus proteins during interferon inhibition of virus particle release. The effect of interferon on the rate of synthesis and the cleavage processing of viral proteins in mouse cells, chronically infected with Rauscher murine leukemia virus, has been studied by immunoprecipitation of newly synthesized viral proteins from virus-infected cells pulse-labeled with [35S]methionine. Immuno-precipitated, labeled polypeptides were resolved by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and then examined by autoradiography. Cleavage processing was studied in the same manner with cells that had been pulse-labeled and then incubated with non-radioactive media for a sufficient time to allow normal cleavage processing to occur. At a concentration that strongly inhibited the release of virus particles, interferon had no effect on the synthesis of proteins carrying antigenic determinants of the major core protein p30 or of the envelope glycoprotein gp69/71. Nor did it affect the post-translational cleavage processing of the precursors to these proteins. Similarly, interferon did not affect labeling or chasing of precursor protein carrying the p15 determinants; labeling of p15 itself could not be studied because it does not contain methionine.


Quantitative granulocyte chemiluminescence in the rapid detection of impaired opsonization of Escherichia coli. Previous work has demonstrated that 40% of clinically isolated Escherichia coli are resistant to in vitro killing by normal human polymorphonuclear granulocytes (PMN) due to ineffective opsonophagocytosis. Using these E. coli isolates, we have demonstrated the usefulness of measuring the chemiluminescence (CL) of granulocytes undergoing phagocytosis in detecting this impaired opsonization. CL correlated well with several other methods to assess PMN function including in vitro killing assays, postphagocytic O2 consumption, and morphological phagocytic indexes. Of the available methods to assess granulocyte function, CL is the most rapid and simple and would appear to be a potentially useful screening method to determine possible opsonic deficiencies of human PMNs and serum. Impaired opsonization to these resistant E. coli isolates was demonstrable in several different donors and could be reversed by type-specific rabbit antibody to the particular resistant isolate. Bacterial levels of superoxide dismutase and catalase, enzymes which have been implicated as possible virulence factors in some microorganisms, did not correlate with resistance in these E. coli isolates. However, heat denaturation of these isolates reversed this resistance and would suggest heat-labile antigenic determinants present in E. coli as possible resistance factors.


Purification and characterization of group A streptococcal T-1 antigen. A method is described for the recovery of purified T-antigen from crude trypsin extracts of an avirulent strain of M-1 protein deficient, T-type 1 group A Streptococcus. The purified T-antigen was resistant to enzymatic degradation with trypsin and pepsin, formed a single precipitin line with standard T-1 antiserum, failed to react with antisera for teichoic acid, group A carbohydrate, and cross-reactive protein antigens, stimulated only a single precipitin system when rabbits were immunized, contained glycine, aspartic acid, glutamic acid, lysine, and serine as the five most predominant amino acids, and consisted of subunit size isomers.


Susceptibility of murine transfer factor to dimerized ribonuclease A. Dialyzable transfer factor was prepared from the spleens of CF1 mice actively sensitized with killed Coccidioides immitis antigen. The transfer factor was administered to normal mice either intraperitoneally or into the hind footpads. The recipient mice were tested for reactivity to the coccidioides antigen and to Candida albicans antigen by means of the footpad swelling test. The transfer factor conferred antigen-specific reactivity upon normal recipient mice when given by the intraperitoneal and footpad routes. This capacity of the transfer factor was destroyed by in vitro pretreatment with dimerized ribonuclease A, an enzyme active against double-stranded, as well as single-stranded, ribonucleic acid. In contrast, monomeric ribonuclease A, which is active against only single-stranded ribonucleic acid under the conditions used here, was without effect upon the transfer factor. These data provide evidence that murine transfer factor contains ribonucleotides that are essential for immunological activity. In addition, the data are consistent with the hypothesis, advanced by others, that the ribonucleotides may be double-stranded or uniquely looped configurations.


Protective capacity of antibodies against Escherichia coli and K antigens. Antibodies to Escherichia coli O and K antigens were raised in rabbits by repeated immunizations with whole, Formalin-killed and, later, liver bacteria. The serum antibody levels were determined with the ammonium sulfate precipitation technique after radioiodinating the antigens. The K antigens had to be conjugated to proteins before labeling. Such conjugations were performed using cyanogen bromide for the K1 antigen and bisdiazobenzidine for the K13 antigen. The protective capacities of the rabbit antisera were tested in intraperitoneally infected mice. The protective capacity of the antisera was expressed per ammonium sulfate precipitation titer. The results showed a significantly higher protective effect for the antibodies against the K1 and K13 antigens than for the antibodies against the O2 and O6 lipopolysaccharides.


Inhibition of leptospiral agglutination by the type-specific main antigens of leptospiras. Microscopic agglutination of leptospiras was inhibited by the homologous type-specific main antigens. The result may indicate that the substance participating in the agglutination of leptospiras with antiserum is the type-specific main antigen.


Identification and solubilization of iodinated cell surface human melanoma associated antigens. To identify soluble cell surface melanoma-associated antigens (MAA), human melanoma cells in culture were radioiodinated by the lactoperoxidase technique and solubilized in non-ionic detergent (NP-40). Labelled MAA were identified by a quantitative double-antibody antigen binding assay and unrelated labelled macromolecules by trichloroacetic acid precipitation. Detergent solubilized 95% of the macromolecule-associated radioactivity. Approximately 8%, presumably MAA, was bound specifically by anti-melanoma serum. In contrast, anti-melanoma serum bound specifically only 0.5 to 1.5% of the acid precipitable radioactivity in control cells iodinated in a similar manner. Specificity was further studied by quantitative serum absorption. Two different melanoma lines were equally effective in inhibiting specific binding of iodinated melanoma lysate, whereas 50-100 times more normal fresh lymphocytes, liver and spleen cells, cultured HeLa or colon adenocarcinoma cells, and 8 times more cultured fetal cells were required to produce similar reductions in specific binding. These studies demonstrate that cell surface human melanoma antigens that differ qualitatively and/or quantitatively from those on normal or malignant allogeneic tissues can be solubilized and identified. These antigens are shared with other melanomas, and some are also present on fetal cells.


Terminal deoxynucleotidyl transferase activity and cell surface antigens of two unique cell lines (NALM-1 and BALM-2) of human leukemic origin. Two unique cell lines, NALM-1 and BALM-2 derived from lymphoblast-like cells of chronic myelogenous leukemia and rare B cell acute lymphoblastic leukemia patients, respectively, were compared with fresh parent cells from the patients and with a Philadelphia chromosome positive K-562 cell line previously established from a chronic myelogenous leukemia patient in blastic phase. NALM-1 resembled the parent cells in the presence of Philadelphia chromosome, non-T/non-B acute lymphoblastic leukemia specific antigens and lack of T or B cell markers, whereas BALB-2, like the parent cells, had two chromosome markers and bore kappa, delta and mu immunoglobulins. NALM-1 lacked Epstein-Barr virus genome, whereas BALM-2 showed the presence of Epstein-Barr virus genome. K-562 cells lacked all the antigen markers examined. All cells had high DNA polymerase alpha activity and low DNA polymerase gamma activity. NALM-1, like the parent cells and unlike K-562 cells, had high terminal deoxynucleotidyl transferase activity of about 200 mu/mg DNA, whereas BALM-2, like its parent cells, had terminal deoxynucleotidyl transferase activity of 1-2 mu/mg DNA (1 u = 1 nmole Mn++-dGTP/h on dA12-18 initiator). Terminal deoxynucleotidyl transferase was characterized by its chromatographic and sedimentation behavior, thermal sensitivity and specific inhibition by streptolydigin and terminal deoxynucleotidyl transferase antisera. These results indicate that NALM-1 and K-562 may represent different phenotypes of cells in CML blastic crisis. Moreover, NALM-1 and BALM-2 seem to have retained the characteristics of original leukemic cells from which they may have been derived.


Expression of alien H-2 specificities of a chemically induced BALB/c fibrosarcoma. The presence of alien histocompatibility antigens on the cell surface of the 3-methylcholanthrene-induced BALB/c (H-2d) fibrosarcoma C-1, was investigated by serological and transplantation studied. Absorption experiments with monospecific alloantisera showed that C-1 cells expressed their original private (H-2.4 and 31) and public (H-2.3, 8, 28, and 35) specificities. C-1 cells were also able to absorb monospecific antisera directed to the private specificity H-2.23 of the H-2k haplotype, as well as antisera to the public specificities H-2.1, 5, 11 11 and 25 (H-2k and in part H-2q, H-2a and H-2b haplotypes), which are absent from H-2d normal cells. Conversely, other alien specificities (H-2.2, 17, 30, 32, and 33) were not detected on C-1 cells. The C-1 cells were also unable to absorb the activity of an anti-Ia serum (1-28) directed to 1a.1, 2 and 19 (lak) specificities. Transplantation studies showed that resistance against the challenge of C-1 cells could be induced in syngeneic BALB/c mice by preimmunization with normal tissues from C3Hf and AKR (H-2k), A (H-2a) and C57BL/6J (H-2b) strains (expressing all or some of the extra H-2 antigens of the tumor) whereas no protection was obtained with DBA/2 (H-2d) or with W/Fu rat tissues. The anti-tumor activity could be passively transferred by BALB/c lymphoid cells immune to normal C3Hf, AKR, A, and NIH (H-2q) tissues, but no protection was achieved with lymphoid cells immune to DBA/2 or to W/Fu normal rat tissues. These data indicate that foreign H-2 antigens are expressed on C-1 tumor and that they might function as tumor-associated transplantation antigen which was shown to be present and individually distinct on this sarcoma by appropriate in vivo tests.


Spontaneous and induced appearance of murine leukemia virus antigen containing cells in organ cultures of embryonic mouse thymus. The potential of embryonic thymus organ cultures for studies on relations of endogenous MuLV, lymphoid cells and thymic microenvironment to lymphoma development were evaluated. Four main findings are reported. First, thymuses of 14-day-old CBA and AKR embryos could be maintained in organ cultures for at least 9 weeks with sustained production of lymphocytes. Lymphopoiesis in CBA and AKR thymuses were not grossly different. Secondly, an indirect immunofluorescence (IF) technique demonstrated spontaneous appearance of MuLV-antigen-containing cells in AKR, but not in CBA thymuses. Such spontaneous MuLV expression first occurred after 16 days of organ culture, thereafter infrequently and at random in individual thymus cultures. Thirdly, incubation of AKR and CBA thymuses in lymphoma extract containing AKR-type MuLV at initiation of organ cultures induced MuLV-antigen-containing cells. These were first detected after 7-14 days in culture, somewhat earlier and initially more frequently in AKR than in CBA thymuses. In the former, induction was accompanied by a clear reduction in the number of lymphocytes per thymus. Fourthly, iododeoxyuridine (IdUrd) treatment of AKR thymuses on cultute day 0, 3 or 7 decreased the number of lymphocytes per thymus and induced appearance of MuLV-antigen containing cells, assayed 8-20 days later. The IdUrd effect was most marked on day 0, and decreased sucessively on days 3 and 7. IdUrd had a much slighter effect on CBA thymuses, causing a lower reduction in cell numbers and inducing few MuLV-antigen cells. These main results clearly demonstrate the potential usefulness of the organ culture system for studied on leukemogenesis. It may be directly applied to answer several questions raised by detailed findings in our study.


Aggregation, dissociation and denaturation of sesame (Sesamum indicum L.) alpha-globulin in cetyl trimethyl ammonium bromide solution. The behaviour of the major protein of sesame seed (Sesamum indicum L.) alpha-globulin has been studied in a cationic detergent, cetyl trimethyl ammonium bromide solution. Up to a critical detergent concentration the protein is precipitated from solution, above which redissolution of the protein is observed. Sedimentation velocity patterns indicate the presence of higher aggregates in the detergent concentration range 5 X 10(-5)--1 X 10(-3) M. These are considered to be the soluble precursors of the insoluble aggregates. Fluorescence measurements show that tryptophanyl groups of the protein which are in contact with the aqueous phase are perturbed by the detergent. The difference spectra of the protein in higher concentration of detergent indicate considerable red shift in the spectrum. Spectrophotometric titration of phenolic groups in 1 X 10(-2) M CTAB indicate that a conformational change in the protein has taken place.


Millin's retropubic prostatectomy. A clinical study. On the basis of two series of retropubic Millin's prostatectomies (426 operations) the value of this method of approach is evaluated. The retropubic procedure seems to offer the best chance for a smooth postoperative course when an open operation is needed.


Complications of transurethral resection of the prostate. A series of 632 patients undergoing one or more transurethral resections of the prostate gland at Wesley Pavilion of Northwestern Memorial Hospital is presented. Group characteristics, associated illnesses, and complications are reviewed and compared to an equally sized group of patients undergoing transurethral prostatic resection at the same hospital some twenty years earlier. This study has shown improvement in incidence of several complications, especially those related to infection and pyelonephritis, and urinary incontinence. Mortality rate has improved significantly and is likely due in great part to more comprehensive care of cardiovascular complications and control of infection. One can perhaps anticipate even further improvements in technique and results with the development of reliable fiberoptic instruments and improved lens systems which have been introduced to our institution since the period of time covered by this review.


[Electron microscopical rapid diagnosis of cutaneous viral diseases]. Electron microscopy by negative staining is a well-known, simple and quick diagnostic method to demonstrate virus particles. A method for routine work is described. The morphology of important viruses of the herpes and pox group is shown. In herpes simplex and zoster, the viruses can be detected in most cases from fresh vesicular fluid, whereas older erosions and crusts are not suitable. Viruses have been shown in all cases of chicken pox, vaccinia inoculata, molluscum contagiosum, and ecthyma contagiosum (orf).


Modification of the acid elution technique for quantitation of fetal hemoglobin in individual erythrocytes. A modification of the acid elution procedure for the demonstration of fetal hemoglobin in red cells using fast green stain is described. This technique not only permits improved visual estimation of the amount of fetal hemoglobin in red cells, but allows for the direct quantitation of fetal hemoglobin content of individual cells, using a scanning and integrating microdensitometer. As an application of this method, the distribution of fetal hemoglobin in populations of red cells of individuals was determined. Distributions were examined in sickle cell anemia patients with different proportions of fetal hemoglobin.


Antibody to the D-alanine carboxypeptidase of Bacillus subtilis does not cross-react with other penicillin-binding proteins. The fact that antibody to d-alanine carboxypeptidase of Bacillus subtilis does not cross-react with other penicillin-binding proteins suggests that these proteins are not precursors or multimers of the enzyme.


Characterization of antiserum directed against form II ribulose 1,5-bisphosphate carboxylase from Rhodopseudomonas sphaeroides. Antiserum directed against form II ribulose 1,5-bisphosphate carboxylase from Rhodopseudomonas sphaeroides showed no cross-reactivity towards the form I enzyme as evidenced by a lack of immunopreciptation. In addition, this antiserum failed to inhibit form I enzymatic activity.


Purification and properties of beta-lactamase from Proteus morganii. The cephalosporin beta-lactamase was purified from a strain of Proteus morganii that showed resistance to beta-lactam antibiotics and produced the enzyme constitutively. The purified enzyme preparation gave a single protein band on polyacrylamide gel electrophoresis and consisted of a single polypeptide of molecular weight 38,000 to 40,000 from gel filtration of Sephadex G-100 and sodium dodecyl sulfate-acrylamide gel electrophoresis, its isoelectric point being pH 7.2 No cysteine residue was found in its amino acid composition. The specific activity was 190 mumol/min per mg of the purified enzyme protein for the hydrolysis of cephaloridine, the optimal pH was about 8.5 and the optimal temperature was 50 degrees C. Antibodies against the purified beta-lactamase inhibited not only the enzyme activity of the purified preparation, but also the enzyme activity of all of the other strains of P. morganii so far tested, regardless of whether the modes of their production were inducible or constitutive. None of the beta-lactamases produced by beta-lactam antibiotic-resistant strains of other species of Proteus was affected at all by the antibodies, thus showing that the purified cephalosporin beta-lactamase was of the species-specific type. The enzymological properties of the preparation have been compared with those of beta-lactamases derived from other gram-negative enteric bacteria.


Immunospecificity of non-histone chromosomal proteins in 89Sr-induced osteogenic sarcoma (mouse). Antibodies against non-histone chromosomal proteins for 89Sr-induced osteogenic sarcoma (mouse) were prepared by immunization of rabbits. The immunoreactivity of this antigen was then compared with those of non-histone chromosomal proteins from Ehrlich ascites tumor, normal mouse liver, and calf thymus by the method of quantitative microcomplement fixation. The non-histone chromosomal proteins of 98Sr-induced osteogenic sarcoma, fractionated by hydroxylapatite chromatography, exhibited significant affinity for the antibodies. Similar proteins from Ehrlich ascites tumor, normal mouse liver, or calf thymus were virtually inactive, indicating the tissue-specificity of 89Sr-induced osteogenic sarcoma proteins.


The RNA-dependent DNA polymerase of avian sarcoma virus B77. Binding of viral and nonviral ribonucleic acids to the alpha, beta2, and alphabeta forms of the enzyme. The extent of binding of various RNA species to the three forms of avian sarcoma virus B77 RNA-dependent DNA polymerase was determined using a sensitive nitrocellulose filter binding technique which was capable of detecting binding reactions with association constants as low as 3 X 10(6) liters X mole-1. All three enzyme forms, alphabeta, beta2, and alpha, bound to all single-stranded RNA species that were tested, including nonviral RNAs. 70 S viral RNA exhibited the highest association constant (about 10(11) liters X mole-1), and a population of virus-derived tRNA molecules from which tRNATrp had been removed, the lowest (about 3000 times lower). The affinity for other RNAs was roughly proportional to their size. The affinity of RNAs for the alphabeta enzyme form always exceeded that for the two others by a factor that depended on the particular RNA, never exceeded 6 and was sometimes as low as 1.2. The association constant of the alphabeta enzyme form with viral 70 S RNA was about 15-fold higher than that with viral 35 S RNA. 35 S RNA annealed to tRNATrp had an association constant that was only 2.5 times higher than that of 35 S RNA alone. This finding suggests that the tertiary structure of 70 S RNA plays a significant role in its affinity for B77 DNA polymerase.