Serotonin nerve terminals in the locus coeruleus of adult rat: a radioautographic study. Serotonin (5-HT) nerve terminals in the locus coeruleus (LC) of adult rat were visualized by high-resolution radioautography, in order to examine their distribution, fine structural features and intimate relationships with norepinephrine neurons. In animals pretreated with a monoamine oxidase inhibitor, prolonged intraventricular perfusion of 10(-4) M [3H]5-HT resulted in a specific identification of most if not all 5-HT axonal varicosities in LC. These terminals were equally distributed between the dorsal and ventral divisions of the nucleus. Their density was approximated at 10(7) per cu.mm within the middle third of the LC. In electron microscope radioautographs, the labeled 5-HT varicosities averaged 0.9 micron in diameter. They all exhibited a distinctive storage organelle, in the form of microvesicles and microcanaliculi (15-25 nm in diameter) partly filled with electron-dense material and usually aggregated in association with several large dense-core vesicles. While this finding of intrinsic morphological characteristics appeared compatible with a special cellular origin or regional differentiation, it was also suggestive of particular functional properties and/or mode of action. In a sample comprised of some 500 sectional profiles from labeled 5-HT varicosities in LC, a small proportion only (less than 10%) exhibited morphologically defined synaptic junctions. These rare contacts were invariably made with dendritic processes and never observed on the noradrenergic perikarya. It is therefore concluded that, in the LC, non-synaptic as well as synaptic mechanisms might be involved in the modulation and transneuronal regulation of norepinephrine neurons by 5-HT afferents.


A comparison of the inhibitory effects of taurine and GABA on identified Purkinje cells and other neurons in the cerebellar cortex of the rat. The microiontophoretic application of taurine and GABA was studied in the cerebellar cortex of the rat. Both taurine and GABA produced a dose-dependent depression of spike frequency of cerebellar neurons. GABA (2-42 nA, mean 27 nA) induced an inhibition of spike discharge on all 138 cells tested, including 29 Purkinje cells. Taurine (60-200 nA, mean 108 nA) induced an inhibition of spike discharge on 93 of the 106 cerebellar neurons tested, including inhibition on 22 of 25 Purkinje cells. Iontophoretic application of bicuculline and picrotoxin antagonized the inhibitory effects of both GABA and taurine on Purkinje cells as well as on cerebellar neurons in general. Strychnine did not antagonize the inhibition of either GABA or taurine. Simultaneous application of taurine and GABA produced a synergistic inhibitory effect on the firing rate of Purkinje cells. Taurine, in contrast to GABA, appeared to be more depressant when applied in the Purkinje cell dendritic zone than when applied near the soma. The data are discussed in terms of taurine functioning as a neurotransmitter in the cerebellum of the rat and having receptor sites distinct from those for GABA.


Tissue culture analysis of neurogenesis: myelination and synapse formation are retarded by serum deprivation. Explants from newborn mouse cerebellum were cultured in nutrient media containing either adequate (30%) or low (15% or 7.5%) serum content. By light microscopic observation, delayed and inhibited myelination was detected in cultures fed with low serum media (experimental cultures). Specific activities of two enzymes related to myelin synthesis, 2',3'-cyclic nucleotide-3'-phosphohydrolase and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were also reduced in experimental cultures. Morphometric analysis of electron micrographs showed that the size of presynaptic endings and total area occupied by synapses in experimental cultures were substantially reduced, while synaptic density per unit area increased. Reflecting the results of synaptic underdevelopment, the levels of two neurotransmitter enzymes, choline acetyltransferase and glutamic acid decarboxylase, were also decreased in experimental cultures.


The levels of apolipoprotein-E in hypercholesterolemic rat serum. The levels of apolipoprotein-C (apo-E) in serum and isolated liproproteins from diet-induced hypercholesterolemic, and to some extent hypertriglycerdemic rats were measured by electroimmunoassay. The hypocholesterolemia was accompanied by a mild hypertriglyceridemia. The apo-E was increased by 60% in the hypercholesterolemic serum with a 5- and 50-fold increase in very low density lipoproteins (VLDL) and low density lipoproteins (LDL) respectively. However, the proportion of apo-E in nascent VLDL isolated from the hepatic Golgi apparatus of hypercholesterolemic rats was significantly decreased. In control serum, 40--50% of the apo-E is found in the density greater than 1.21 g/ml fraction, although this is at least partially due to ultracentrifugation. The aproprotein is absent from the density greater than 1.21 g/ml fraction from hypercholesterolemic serum, suggesting that it is bound more firmly to the lipoprotein complex. It is concluded that the large increases in apo-E in the VLDL and LDL density ranges of serum from hypercholesterolemic rats may in part be accounted for by the utilization of apo-E normally found at higher densities.


Comparison of rotavirus strains by hemagglutination inhibition. Rotaviruses have been shown to be of importance as aetiologic agents of gastroenteritis in infants and in domestic animals of several species. Hemagglutinins were prepared from two Canadian isolates of bovine rotavirus and from one isolate of a simian rotavirus. A United Kingdon isolate of bovine rotavirus was shown not to possess hemagglutinating activity, indicating a strain difference between a Canadian and United Kingdom bovine rotavirus. In hemagglutination-inhibition (HAI) tests a rabbit hyperimmune (two injections) serum, prepared to one of the bovine rotaviruses, was not helpful in distinguishing the two bovine viruses because of cross-reactions between the viruses. However, it was possible to distinguish the bovine viruses from the simian virus with this serum. When guinea pig immune sera were prepared to the four rotavirus strains and tested with the three hemagglutinins in the HAI test, antigenic differences between the four strains of rotavirus were demonstrated. Hyperimmune guinea pig serum prepared to a strain of human rotavirus did not inhibit any of three hemagglutinins indicating that the human strain is different from the three rotavirus strains which gave hemagglutinins.


Inhibition of protein synthesis in Streptococcus faecalis by ochratoxin A. A non-competitive inhibition of binding of cAMP to bovine protein kinase by ochratoxin A (OTA) is shown. Preliminary evidence of a protein kinase in Streptococcus faecalis is presented. The cAMP stimulation of this kinase is also inhibited by OTA. At the lowest OTA concentrations, RNA and protein synthesis are inhibited in S. faecalis. The inhibition of RNA synthesis is secondary, as in the presence of chloramphenicol no inhibition occurs for 10 min after the addition of OTA. The synthesis but not the induction of beta-P-galactosidase is inhibited by OTA. The polysomes of S. faecalis are stabilized after addition of OTA, showing an inhibition of peptide elongation. The model of action of OTA in bacteria is discussed and it is concluded that inhibition of protein synthesis is the process which might be closest to the primary target of OTA.


Metastatic cystosarcoma phyllodes. A report of 2 cases presenting with neurological symptoms. Two cases of cystosarcoma phyllodes of the breast are presented with central nervous system (CNS) metastases appearing several years after mastectomy for the primary lesion. Unusual features in these cases include the widespread metastases themselves, neurologic symptomatology and CNS involvement, metastases into a uterine leiomyoma and into an area of hepatic adenomatous hyperplasia, and glomus-like structures in one of the primaries with similar structures resembling glomus cells in metastases. The distant metastases were of stromal cells only and frequently surrounded epithelial cells to isolate the indigenous glandular structures. The concept that cytosarcoma is a peculiar stromal neoplasm rather than a tumor of dual neoplastic origin is discussed.


Histologic grading of Wilms' tumor as potential prognostic factor: results of a retrospective study of 26 patients. A histologic grading system based on tumor differentiation was applied in a retrospective study of 26 patients with Wilms' tumor to determine if it might provide an index to prognosis. The results were compared to those obtained by applying a histologic classification based on the presence or absence of individual histologic structures to the same tumors. Low grade tumors with predominance of differentiated structures--glomeruli and tubules--were associated with a better cure rate than high grade tumors composed mainly of undifferentiated spindle elements. The presence of undifferentiated large cells correlated with poor cure rate. The findings suggest that histologic grading may be valid as a prognostic factor in Wilms' tumor.


An immunologic basis for detection of occult primary malignancies of the head and neck. After extensive evaluation of patients with metastatic neck disease and clinically undetectable primary cancer of the head and neck, the clinician is often faced with the difficult question of subsequent management. In this study, sera from 11 patients with clinically occult carcinoma and metastatic lymphadenopathy were studied for Epstein-Barr virus-associated antigens. These were compared with 35 sera from patients with known nasopharyngeal carcinoma at all stages of disease and treatment and with 212 sera from control patients with other head and neck tumors, patients with lymphoma, and normal controls. There was a significant correlation between high antibody titers to Epstein-Barr virus, especially in the serum IgA fraction, and the presence of nasopharyngeal carcinoma. Thus, identification of occult nasopharyngeal carcinoma by immunologic means may have important application in the selective management of the patient with an unknown head and neck primary malignancy.


Hepatic artery ligation and adriamycin infusion chemotherapy for hepatoma. Two patients with hepatoma of the liver were treated with ligation of the hepatic artery and continuous infusion of adriamycin into the distal arterial branch. Both patients had tumor replacing 60-75% of the normal liver parenchyma. There was clinical improvement and objective shrinkage of the tumor by liver scan lasting for 5 and 7 months. But both cases died 7 and 11 months later, probably of cardiac toxicity (total dose of adriamycin = 500 mg/m2).


Inflammatory fibrous histiocytoma: case report. This report concerns a patient with inflammatory fibrous histiocytoma, who in contrast to previous reported cases, has had a long survival (20 years), without evidence of recurrent disease following treatment. An interesting but nonreproducible study was the development of leukemia in 2 of 3 Swiss strain mice following the intraperitoneal injection of a saline extract of the patient's tumor.


Wilms' tumor: prognostic factors for patients without metastases at diagnosis: results of the National Wilms' Tumor Study. Multivariate statistical analyses are used to evaluate a wide range of factors in predicting relapse and survival for 429 children enrolled in the National Wilms' Tumor Study. Anaplastic or sarcomatous histology, specimen weight over 250 grams, positive regional lymph nodes, treatment with only a single drug and age over two years are the most important predictors of relapse. The first three factors also predict mortality. Laterality, capsular penetration, intrarenal vascular invasion, direct regional extension and operative spillage have lesser effects which do not contribute significantly to the multivariate prediction equations, while sex, race and the presence of a tumor thrombus in the renal vein have essentially no effect. Treatment with combination chemotherapy is confirmed to be efficacious regardless of what prognostic factors are operating. A statistical model allows prediction of relapse and death for each of 24 patient subgroups, with the estimates of percentage relapsed ranging from 3% to 100%.


Ocular granulocytic sarcoma (chloroma) with acute myelomonocytic leukemia in Turkish children. In a series of 166 leukemic children from Turkey, 56 had acute myelomonocytic leukemia (AMML). Seventeen boys and 3 girls presented with chloroma-like deposits (granulocytic or myeloid sarcomas) in the eye and orbit, all showing AMML on initial study of blood and marrow. The ocular lesions responded rapidly to antileukemic therapy. Laboratory studies of AMML cases revealed no cytogenetic or immune defects, and Epstein-Barr virus titers were normal. A group-specific (GS-3) antigen (type-C virus?) was identified in one patient by radioimmunoassay of orbital tumor extracts. It is not clear what factors contribute toward the myelomoncytic differentiation of leukemia and its localization in the eye and orbit, but opportunities for further study are enhanced by reports of a predisposition to ocular chloroma among leukemic children in Africa, Egypt, and Japan.


Abrogation of tumor rejection by trypan blue. Trypan blue treatment prevented tumor-specific rejection in three animal model systems. These included the spontaneous rejection of tumors of mice (UVT-2051) and guinea pig (line 1 hepatoma) as well as vaccine-induced rejection of a guinea pig tumor (line 10 hepatoma). Secondary immune reactions to line 10 cell challenges were not abolished by trypan blue treatment. Although trypan blue is a potent inhibitor of macrophage cytotoxicity in vitro, the mechanism by which it inhibited tumor-specific rejection has not been established.


Chromatin-associated glycoproteins of normal rat liver and Novikoff hepatoma ascites cells. Glycoproteins were demonstrated in the 0.6 M NaCl extract of chromatin of normal liver and Novikoff hepatoma cells by periodic acid-Schiff staining of sodium dodecyl sulfate-polyacrylamide gels. In chromatin extracts of Novikoff hepatoma cells, six major periodic acid-Schiff-positive bands coincident with Coomassie Blue-stained protein bands were found with molecular weights of 30,000, 54,000, 64,000, 75,000, 104,000, and 127,000. A corresponding extract from normal rat liver chromatin revealed the presence of four major periodic acid-Schiff-positive bands that migrated with protein bands at molecular weights of 16,000, 30,000, 54,000, and 75,000. The glycoproteins of these extracts contained either mannose or alpha-D-glucose, fucose, and N-acetylglucosamine as shown by the specificity of lectins in affinoelectrophoresis. One of the glycoproteins detected by affinoelectrophoresis was very basic.


Production of plasminogen activator by cells transformed by herpesviruses. Plasminogen activator is produced by hamster cells transformed by human herpesviruses. These cell lines have previously been shown to be oncogenic when injected s.c. into newborn syngeneic hamsters. Lysis of fibrin overlays by these cell lines was plasminogen dependent. Normal hamster embryo fibroblasts and a hamster cell line transformed by PARA-7 (an adenovirus-SV 40 hybrid) failed to produced lysis. In separate experiments fibrin overlay of lytically infected secondary rabbit kidney cells did not show induction of this activity during the normal course of productive infection. The human cell line TE-85 clone F-5, a clonal cell line from a human osteogenic sarcoma, failed to produce plasminogen activator, but two separate clones of these cells that were morphologically transformed after exposure to UV-inactivated herpes simplex virus type 2 produced rapid lysis of the fibrin overlay. Clonal variation was observed in herpes simplex virus types 1 and 2-transformed hamster lines and is under investigation. It is suggested that plasminogen activator detection may serve as a convenient assay system for transformation of normal cells by herpesviruses.


Marked stimulation of lymphocyte-mediated attack on tumor cells by target-directed liposomes containing immune RNA. A marked stimulation of normal guinea pig lymphocytes was obtained by incubating them with liposomes that contained both antibody to lymphocytes to provide "homing" of the vesicles and immune RNA isolated from guinea pigs immunized with syngeneic line 10 hepatocarcinoma cells. Tumor cell cytotoxicity was monitored by a 51Cr release assay. This target cell delivery of immune RNA by liposomes produced a dose-dependent stimulation up to 12 times that achieved by in vitro methods with naked immune RNA.


Electron microscopic studies of intracisternal virus particles in Soehner-Dmochowski murine sarcoma virus-induced bone tumors of New Zealand Black rats. Soehner-Dmochowski murine sarcoma virus (Moloney)-induced bone tumors of New Zealand Black rats carry two morphologically different types of virus particles, namely, extracellular type C and intracisternal virus particles, which have thus far not been reported. These two types of virus particles have also been observed in the tissue culture cells derived from normal prostate tissues of A/Dm and BALB/c/Dm mice after inoculation of cell-free extracts of these bone tumors. The intracisternal virus particles, 90 to 120 nm in diameter, have always been found in the rough endoplasmic reticulum; they have two inner concentric layers with a relatively electron-lucent center, frequently showing cylindrical, chain-like, or multipolar budding forms. Type C virus particles produced by Soehner-Dmochowski murine sarcoma virus (Moloney)-infected prostate tissue culture cells from A/Dm and BALB/c/Dm mice belong to the murine sarcoma-murine leukemia virus group, as revealed by the fixed immunofluorescence test and by immunoelectron microscopy. The morphological and immunological relationship of intracisternal virus particles and other types of virus particles (such as type C, type H, and intracisternal type A virus particles) and intracisternal virus particles in guinea pig leukemia are defined by routine electron microscopy observations and by immunoelectron microscopy studies.


Prevalence of non-T-cells in the replication of the N-tropic, type C virus of young AKR mice. The XC infectious center assay was used to study the nature of the lymphoid cells producing N-tropic C-type viruses in preleukemic AKR mice. Viral production by thymic cell suspensions was very low and was possibly due to contaminating cells. Production at least 100-fold higher was found in spleen cells and was probably due to non-T-cells. The significance of these results is discussed briefly, including the possibility that the N-tropic XC syncitia-inducing type C virus of young AKR mice is not the leukemogenic agent.


Abnormal distribution of O-alkyl groups in the neutral glycerolipids from human hepatocellular carcinomas. The direct comparison of O-alkylglycerol composition in the neutral lipids fractions prepared from human hepatocellular carcinomas with that in corresponding preparations from nonneoplastic liver is reported. Tumor-bearing liver and noncancerous liver specimens were obtained either during surgery or at autopsy. Thirty different tissue specimens obtained from 18 cases were analyzed. Representative samples from each specimen were examined microscopically to confirm the pathological diagnosis. Gas chromatographic analysis of alkylglycerol derivatives showed that hexadecylglycerol, octadecylglycerol, and octadecenylglycerol were the principal components. Compared to the noncancerous liver, hepatocellular carcinomas contained higher proportions of hexadecylglycerol and lower proportions of both C18-glyceryl ethers. Associated with this change was an increase in the proportion of saturated to monoenic alkylglycerols. These abnormalities appeared to be more severe in the necrotic areas of the tumors. Higher concentrations of neutral alkyl glycerolipids and of cholesterol were found in the tumors; no differences between the two groups could be found in the levels of ether-linked phosphoglycerides, triglycerides, and lipid phosphorus.


Induction of sister chromatid exchanges by transformation with simian virus 40. The frequency of sister chromatid exchange (SCE) has been followed sequentially after the addition of SV40 to human diploid fibroblast cultures. The SCE frequency was nearly the same in uninfected controls and in infected cultures before they became tumor antigen positive. When cells exhibited tumor antigen, the SCE frequency increased over a wide range, and changes in chromosome number and structure were observed simultaneously. Cells with induced chromosome abnormalities without increased SCE's and the reverse present the possibility that the two phenomena have different viral mechanisms. This increase in SCE can be added to the previously demonstrated change in chromosome number and increase in chromosome breakage and rearrangement as indicators of genetic damage associated with viral transformation.


An epithelial cell line with chronic polyoma infection established from a spontaneous mouse pancreatic adenocarcinoma. The establishment of an epithelial cell line from a mouse pancreatic adenocarcinoma is described. The cell line, designated LTPA, was aneuploid and exhibited many transformed growth properties (rapid growth rate, failure to show density-dependent inhibition of growth, ability to grow in defined medium). A type C oncornavirus was isolated from the culture medium, and electron microscopy also revealed the presence of intracisternal type A particles. LTPA cells carried a persistent polyoma infection which produced only low levels of cytopathic effects. A mycoplasmal contamination was also carried. When injected s.c. into Swiss nu/nu mice, LTPA cells formed ductular structures which were destroyed by inflammatory reactions within 3 weeks.


Increased susceptibility to feline leukemia virus infection in cats exposed to methylnitrosourea. Exposure of adult specific-pathogen-free cats to methylnitrosourea resulted in increased susceptibility to infection by feline leukemia virus. A greater proportion of cats exposed to methylnitrosourea and feline leukemia virus (69%) became persistently viremic than those exposed to feline leukemia virus alone (17%). Segmented neutrophils were reduced by 90 to 99% within 3 days following exposure to methylnitrosourea, (15 to 20 mg/kg) whereas the effects on lymphocytes and erythrocytes, although less obvious, were also detected.


Growth of transplanted rat tumors following administration of cell-free tumor antigens. The variable effect of immunizations with cell-free tumor extract (TE) in a weakly antigenic (Morris hepatoma 3924a) and a highly antigenic (glioma 9L) inbred rat model system was described. Tumor enhancement was noted with the weakly antigenic tumor following either immunization with a low dose of TE admixed with Freund's adjuvant or immunization with high doses of TE. Enhancement was observed at four tumor cell challenge levels. Tumor enhancement with the highly antigenic tumor was found at only one level and was seen following immunization with intermediate doses of TE and a high tumor cell challenge. Tumor protection was only detected with the weakly antigenic tumor following immunizations with an intermediate dose of TE and challenge with a tumor cell dose 5-fold over threshold. This protection was tumor specific, as judged by an amputation and rechallenge experiment.


Calcium requirements for the proliferation of cells infected with a temperature-sensitive mutant of Rous sarcoma virus. Between 32 degrees and 39 degrees, uninfected normal rat kidney (NRK) cells proliferated minimally, or not at all, in medium containing 0 to 0.01 mM extracellular free calcium. Transformation of NRK cells by wild-type avian sarcoma virus B77 enabled them to proliferate equally rapidly at all temperatures in high (1.25 mM)- or low (0.001 mM-calcium medium. Transformation by the temperature-sensitive LA-23 mutant of Rous sarcoma virus also enabled NRK cells to proliferate in low (0.001 mM)-calcium medium at 32 degrees or 37 degrees. However, the LA-23 NRK cells could not proliferate in low (0.001 mM)-calcium medium at a nonpermissive temperature such as 39 degrees, which prevents expression of other aspects of the transformed phenotype.


Intratumor chemoimmunotherapy with mitomycin C and components from mycobacteria in regression of line 10 tumors in guinea pigs. Intratumor chemotherapy with the use of mitomycin C and/or immunotherapy caused regression of line 10 carcinomas in strain 2 guinea pigs and resulted in development of tumor-specific immunity. The immunotherapeutic preparation consisted of oil-in-water emulsions containing Mycobacterium cell walls or residues of cell walls termed cell wall skeletons. The latter preparations were combined with trehalose dimycolate, which was isolated by microparticulate chromatography from whole cells of mycobacteria. Reducing mitomycin C to a single intratumor injection of 50 microgram produced little necrotizing effect and a mean tumor regression rate of 17%. Intratumor immunotherapy 1 day after treatment with 50 microgram of mitomycin C resulted in regression of 90% of the treated tumors as compared to mean regression rates of 30 to 50% for immunotherapy alone. In addition, chemoimmunotherapy was more effective than either chemotherapy or immunotherapy alone in producing regression of relatively large, as well as smaller, tumors.


Activities of key gluconeogenic enzymes and glycogen synthase in rat and human livers, hepatomas, and hepatoma cell cultures. The activities of pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G6Pase), and glycogen synthetase (GS) were determined in the cancerous and in the apparently uninvolved (host) regions of livers from primary hepatoma patients as well as in normal adult human livers and human fetal livers. The activities of these enzymes were also assayed in a fairly fast-growing, 3'-methyl-4-dimethylaminoazobenzene-induced transplantable rat hepatoma and in hepatoma cell lines derived from both rat and human tumors. In the human hepatoma, as in the rat hepatoma, the activities of PC, PEPCK, and G6Pase were considerably reduced, compared to those in the host liver. The activities of both the a (glucose 6-phosphate-independent) and b (glucose 6-phosphate-dependent) forms of GS were also lower in human and rat hepatomas than in the respective host livers. Activities of PC, PEPCK, and G6Pase in the human hepatomas were often comparable with those of fetal livers. In rat and human hepatoma cells, the activities of PC, PEPCK, and G6Pase were similar to or lower than the activities in the respective hepatomas; the activities of GS a were also similar to those in the hepatoma, whereas the activities of GS b were somewhat higher.


Alteration of sodium transport in mouse mammary epithelium associated with neoplastic transformation. Using electrophysiological techniques we have examined the apical membrane ionic permeabilities of primary cell cultures of the mouse mammary gland in the midpregnant, preneoplastic, and neoplastic states. Membrane Na+ permeability changed with tumorigenesis, whereas K+ and Cl- permeabilities were unaltered. With tracer flux techniques the unidirectional efflux rate constant of 22Na was found to be greater in tumor cells than it is in normal cells. This increase in 22Na efflux was eliminated by the addition of ouabain. The results are interpreted as an increase in Na+ permeability and in Na+-K+-ATPase activity with the neoplastic transformation. The presence or absence of the virus in midpregnant cells does not seem to affect Na+ permeability.


Characterization of a unique cell line (LAZ 221) from human acute lymphocytic ("null" cell) leukemia. A unique human cell line designated LAZ 221 has been established from the peripheral blood of a patient with acute lymphocytic leukemia of the "null" cell type. The cell line does not possess the Epstein-Barr virus nuclear antigen and has a karyotype of 45,XX,-9,-12,+(9q12q). Both the established cell line and the patient's uncultured blast cells share the same phenotypic markers. They both lack T-cell markers. They fail to form sheep erythrocyte rosettes and do not react with T-cell-specific antisera (TH1-, HTL-), nor do they possess B-cell markers. They do not form rosettes with erythrocytes sensitized with complement, and they are surface immunoglobulin negative. However, they do possess an HLA-D-related glycoprotein complex of 23,000 to 30,000 daltons, an la-like antigen. Thus, LAZ 221 shares the phenotype of the patient's uncultured blasts and is a cell line representative of about 75% of all human acute lymphocytic leukemias. In this respect it differs from previously described human hematopoietic cell lines.


Immunization of mice against murine mammary tumor virus infection and mammary tumor development. Formalin-inactivated whole murine mammary tumor virus (MuMTV), VuMTV membranes, the acid-soluble component of MuMTV, and purified MuMTV glycoprotein with a molecular weight of 55,000 (gp55; also designated as gp52) were used as vaccines in an attempt to identify the MuMTV antigen(s) that can protect mice from exogenous MuMTV infection and subsequent tumor development. Formalin-inactivated whole MuMTV, MuMTV membranes, and purified MuMTV gp55 were effective immunogens, whereas the acid-soluble component of MuMTV (which consists mainly of MuMTV gp55) failed to protect mice from challenge with live virus. These results suggest that (a) MuMTV gp55 is the major immunizing antigen and (b) its native conformation must be maintained for it to be an effective vaccine.


Fine structure of crystalline inclusions in B-cells of the islets of Langerhans in the alligator. The ultrastructure of crystalline beta granules of the islets of Langerhans in the alligator has been investigated. From optical diffraction analysis and serial sectioning, the existence of four distinct types of crystalline inclusions was established in ultrathin sections. The first type is the most frequent and is interpreted as a rhombohedron with a base, the ortho-hexagonal unit-cell edges being a equal to 18.9 nm, c equal to 23.0 nm. The second type of crystal (not observed in serial sections) is found compatible with a rhomb-dodecahedron which indexes on a cubic cell with a equal to 9.6 nm. The third type of crystal was assigned to dipyramids. Dipyramids are extremely rare, and only two diffraction patterns were obtained; their crystal system could not be determined. Prisms, which are second in abundance, represent the fourth type of crystal. Spacings as well as the symmetry differ from those of the above three crystal types and indicate a tetragonal cell with a equal to 4.2 nm, c equal to 14.2 nm. The data for the prismatic crystals are strikingly similar to those of proinsulin and may represent the first case of agreement between crystals (i) formed in vitro and studied by X-ray diffraction and (ii) those investigated in situ by electron microscopy.


Lack of effect of receptor blockers on the formation of long-lasting associations between sympathetic nerves and cardiac muscle cells in vitro. Sympathetic nerves in vitro form long-lasting, intimate, functional relationships with cardiac muscle cells, but not with fibroblasts. In the presence of an adrenergic beta-blocker and a cholinergic muscarinic blocker, long-lasting relationships still take place. It was concluded that neurotransmitter "receptors" are not involved in the mechanism of "recognition" of cardiac muscle cells by sympathetic nerves.


Evidence for coupled synthesis of mRNA for ribosomal proteins and rRNA. Two-dimensional gel electrophoresis revealed quantitative differences in the 35S-methionine labeled proteins synthesized in vitro in the wheat germ system containing poly A(+) mRNA of normal liver, regenerating liver or Novikoff hepatoma cells. A group of low molecular weight proteins which comigrated with proteins of 40S ribosomal subunits were synthesized in a much greater concentration with the poly A(+) mRNA from the latter two sources. This correlates with increased synthesis of ribosomal proteins in regenerating liver or tumors which is temporally coupled with the increased nucleolar synthesis of rRNA precursors.


Periodate induced cross-linking of concanavalin A-reactive membrane proteins of rabbit thymocytes. Purified plasma membranes of rabbit thymocytes are exposed to sodium periodate and galactose oxidase at conditions similar to those used to induce mitogenic transformation of lymphocytes. The membrane proteins are then fractionated by dodecyl sulfate poly-acrylamide gel electrophoresis. At concentrations of 0.005 M, Na IO4 cross-links 55,000 D and 110,000 D glycoproteins which are known to specifically bind concanavalin A. Galactose oxidase has a similar cross-linking effect, but, at the same time causes proteolytic degradation of membrane proteins. Our data indicate that oxidizing agents, like NaIO4 and galactose oxidase, can indeed cross-link receptors of the thymocyte plasma membrane as has often been proposed as a possible mechanism of their action.


The effect of experimental uremia on potassium-activated phosphatase from erythrocyte and cardiac membranes. Unilateral nephrectomy and partial resection of the contralateral kidney in rats causes uremia that lasts for several weeks and influenced overall (Na+, K+)-ATPase activity as well as p-nitrophenyl phosphatase (p-NPPase) in the plasma membranes of erythrocytes and heart. Both enzymatic activities are inhibited to a similar extent in the uremic animals, regardless of whether the membranes were investigated one or four weeks after surgical treatment.


Detection of Fabry's disease heterozygotes by hair root analysis. The alpha-galactosidase/beta-hexosaminidase ratio was measured for individual hair roots as a method for heterozygote detection in Fabry's disease. Hair root analysis in control individuals revealed no striking sex difference in alpha-galactosidase/beta-hexosaminidase ratio when five males and five females were compared. The values for the ratio X 100, calculating both enzyme activities in nmol of product per min per microliter of hair extract, ranged from 0.8 to 9 for controls and from less than 0.1 to 0.4 for two hemizygous males. Hair root analysis in four heterozygotes with clinical evidence of disease gave values for each individual in the control range, in the range for hemizygotes and in an intermediate range. The experience using hair root analysis for heterozygote detection in the X-linked Lesch-Nyhan syndrome suggests that this approch will be a sensitive heterozygote detection method which takes advantage of the occurrence of hairs with a deficient phenotype on the basis of Lyonization. We observed an affected male who was born to a female without clinical or biochemical evidence (examination included extensive hair root analysis) of Fabry's disease, thus documenting a likely instance of new mutation.


Calcitriol in dialysis patients.

Calcitriol in dialysis patients. We conducted a 7-month randomized, single, double, single-blind comparison of calcitriol (1,25(OH)2D3) with vitamin D3 in 22 hemodialysis patients to study the effects on the biochemical abnormalities associated with osteodystrophy. Calcitriol was given for 3 mo. All patients had initial prestudy calcium values less than or equal to 9.5 mg/100 ml, and phosphate values less than or equal to 4.5 mg/100 ml. Data were analyzed using the Normalized Trend Index (NTI). Calcitriol induced a rise in calcium (8.7 to 10.25 mg/100 ml) (p less than 0.001) and a fall in alkaline phosphatase (p less than 0.005), while D3 had no appreciable effect. The mean dose of calcitriol during treatment was 0.579 microgram/day while that for D3 was 706 IU/day. The effect on serum phosphate concentration was variable. Hypercalcemia as high as 13.2 mg/100 ml occurred in 2 of 13 patients on 1,25(OH)2D3, but in every instance promptly returned to normal with dose reduction. No other adverse effects were noted with therapy. We conclude that calcitriol reverses the biochemical abnormalities of osteodystrophy. Since its effects are rapidly reversed with discontinuation, the drug is probably safe as well as effective.


Radiology of needle biopsy of pulmonary lesions. A technique for percutaneous aspiration needle biopsy is described and discussed. It is concluded that it forms a simple and easy way of obtaining histological material and it can be carried out in most radiological departments.


Lecithin--cholesterol acyltransferase and the lipoprotein abnormalities of obstructive jaundice. 1. We have studied the plasma lipoprotein abnormalities in obstructive jaundice to test the hypothesis that the abnormalities would correlate with plasma lecithin--cholesterol acyltransferase activity. 2. Very-low-density lipoproteins (VLDL) were normal in composition and had a normal pre-beta electrophoretic mobility when lecithin--cholesterol acyltransferase activity was high. When it was low VLDL had abnormal composition and ran in the beta position. 3. With high lecithin--cholesterol acyltransferase activity, low-density lipoprotein (LDL) was normal and cholestatic LDL (LP-X) was not found. With low lecithin--cholesterol acyltransferase activity up to three LDL fractions were found: (i) large triglyceride-rich particles, (ii) LP-X and (iii) a triglyceride-rich cholesteryl ester-poor particle of normal dimensions. 4. High-density lipoprotein (HDL) concentrations correlated with lecithin--cholesterol acyltransferase activity and HDL were normal in composition and electron-microscopic appearance when the activity was high. When lecithin--cholesterol acyltransferase activity was low HDL was abnormal in composition and 'stacked discs' were seen on electron microscopy as well as normal spherical particles. 5. These results suggest that low lecithin-cholesterol acyltransferase activity may cause at least some of the lipoprotein changes of obstructive jaundice.


Effect of antitonin on blood pressure in the one-kidney hypertensive rat. 1. The tonin concentration of saliva and submaxillary glands was studied in one-clip hypertensive rats with or without the contralateral kidney. 2. Salivary tonin concentration was elevated in one-kidney hypertensive rats, but not in one-kidney normotensive or two-kidney, one-clip hypertensive rats. In contrast, an elevated submaxillary gland tonin concentration was found only in uninephrectomized animals, whether normotensive or hypertensive. 3. A single intravenous administration of rabbit tonin antiserum into one-kidney hypertensive rats restored blood pressure to normal in seven out of ten animals. There was little change in blood pressure in two-kidney, one-clip hypertensive, uninephrectomized or sham-operated rats. 4. These findings suggest a connection between the physiology of the kidney and of the submaxillary gland in the rat, and indicate that tonin may play a significant role in maintaining high blood pressure in one-kidney hypertensive aniamls.


The importance of adrenocortical steroids in the pathogenesis of ketosis in diabetic guinea pigs (Caviae porcellus). The roles of adrenocortical steroids in the pathogenesis of diabetic ketosis were investigated in guinea pigs. Groups of normal and Streptozotocin-induced diabetic animals were either given saline as a control, methylprednisilone as an exogenous steroid, or adrenocorticotrophin to stimulate endogenous steroid production. Plasma and urine ketone determinations were made daily for six days at which time animals were necropsied. The qualitative procedure used failed to detect ketonemia, and ketonuria was only found in diabetic guinea pigs treated with steroids. The results support the hypothesis that the absence of ketosis in spontaneously diabetic guinea pigs may be the result of a relative glucocorticoid insufficiency secondary to fatty degeneration of the adrenal glands, a common lesion in spontaneously diabetic guinea pigs.


Quantitative ESR-studies of decay-produced radicals in 5-iododeoxyuridine labeled with 125I, 131I or tritium: role of the Auger effect. In view of the enhanced biological damage caused by the "Auger nuclide" iodine-125, we have carried out quantitative ESR-studies of the radical formation in polycrystalline 5-iododoeoxyuridine (IUdR) resulting from the following internal or external radiation sources: (1) Decay of 3H, 125I or 131I in labeled IUdR; (2) Lanthanum K-photons corresponding to the K-edge of iodine; (3) 60Co gamma-rays. The results clearly indicate that inner shell ionization with its accompanying Auger effect as caused by the lanthanum K X-ray produces about 30% more free radicals per unit dose absorbed than 60Co gamma-rays, when considering the long-lived secondary radicals. Similarly, the concentration of free radicals is by about 30% higher in 125I- than in 131I-labeled IUdR at comparable doses. In the case of 3H-labeled IUdR the dose curve is almost identical with that observed for 125I-labeled IUdR. The results are discussed in terms of a localized radiation damage from low energy electrons.


The swollen limb: cutaneous clues to diagnosis and treatment. We have reviewed a wide spectrum of disorders producing a swollen limb, some common, others uncommon. Cutaneous signs are often seen in association with these conditions and careful attention to them by the dermatologist may be helpful in determining an accurate diagnosis and appropriate therapy.


Fine structure of wall bodies in large cells of an Anabaena species. A new type of inclusion in blue-green algae termed a wall body is described. These are elongate bodies of medium electron density, from 6 to 70 nm in thickness and at least 0.2 micrometer in depth found in large cells of Anabaena sp. B387. The bodies are up to 12 micrometer long, extending the complete length of the cell in some cases. Some of the bodies are bifurcated and the ends appeared to come in contact with the plasma membrane. The wall bodies lie at various angles in the cells. Up to six have been observed in a single thin section of a cell. The large cells are found in greater numbers in cultures grown in a medium solidified with agar. Comparison of the bodies to the mucopolymer layer (layer 2) of the cell wall is made.


Yunnan Bai Yao-induced platelet release in suspensions of washed platelets. The effects of Yunnan Bai Yao on platelets was investigated in the absence of plasma co-factors. Platelets were washed with Tyrode's solution and incubated with this herbal drug at 37 degrees C. It was found that this drug can, to a lesser extent, also induce the release of platelet constituents in the absence of the plasma co-factors. The possible action of this drug is also discussed.


Endocrine and pharmacological factors which influence the onset of labour in rhesus monkeys. Indomethacin administration in late pregnancy prolonged gestation in caged rhesus monkeys and inhibited premature labour and postponed delivery in chronically catheterized monkey fetuses. Chronic indomethacin treatment was associated with a reduction in the urinary excretion of a prostaglandin metabolite, a potent inhibitory effect on myometrial cyclic AMP phosphodiesterase, and severe oligohydramnios in pre-term and post-term fetuses. Experimental anencephaly (functional hypophysectomy) of the rhesus fetus results in lowered concentrations of maternal oestradiol and loss of the precise control of gestational length, with 40% of fetuses delivering beyond term. Corticotropin (ACTH) infused into the fetus results in raised concentrations of fetal and maternal cortisol, progesterone and oestrogens. Progesterone concentrations in peripheral blood apparently have little bearing on uterine quiescence in the rhesus monkey, since the concentrations of progesterone in maternal and fetal blood vary directly with uterine activity. The results of chronic infusion of corticotropin in the fetal monkey support the theory that in the monkey parturition is mediated by increased oestrogen production by the fetoplacental unit and by a rise in the concentrations of oestrone and prostaglandin in the amniotic fluid.


The fetal hypothalamus and pituitary in the initiation of labour. Evidence is presented for a number of events in the fetal hypothalamic-pituitary axis which may play a key role in the onset of labour: (1) In the sheep fetus a progressive rise in the fetal circulating concentrations of corticotropin in the days preceding delivery; (2) In the human fetus a switch from the production of corticotropin-like fragments (melanotropin and corticotropin-like intermediate lobe peptide) to authentic corticotropin in the last weeks of gestation; there is evidence also for a placental origin of corticotropin; (3) In the human fetus, a release of oxytocin and vasopressin associated with the process of spontaneous labour.


Fetal cortisol and the initiation of labour in the human. The role of the fetal adrenal activity in the initiation of parturition in the human has been investigated. Women were studied in the last trimester of pregnancy during treatment with betamethasone for prevention of the idiopathic respiratory distress syndrome. Although betamethasone caused a considerable drop in the cortisol concentrations of fetal plasma and amniotic fluid, the time to spontaneous delivery in this group was similar to that in matched controls. Moreover, cortisol in fetal scalp blood at the onset of parturition in untreated women did not differ between those with spontaneous and those with induced labour. Evidence is given that increase of circulating cortisol in the fetus during the course of parturition predominantly reflects a rise in maternal cortisol under the influence of labour. The strain of labour seems to partly override the betamethasone-induced inhibition of maternal cortisol release. The possible ability of the fetal adrenals to respond to stimuli is illustrated by comparison of cortisol concentrations in cord plasma after various forms of complicated deliveries.


Parturition in the sheep.

Parturition in the sheep. Parturition in sheep is initiated by a sharp rise in the rate of secretion of cortisol by the fetal adrenal. Increased secretion is due partly to enhanced responsiveness to corticotropin (ACTH) and partly to increased fetal concentrations of corticotropin. Cortisol acts on placental enzymes active in the biosynthesis of oestrogens from progesterone. Thus placental secretion of oestrogen increases and that of progesterone decreases. This change in the ratio of oestrogen: progesterone, particularly the rise in oestrogen, stimulates release of prostaglandin F2alpha (PGF2alpha) from the maternal placenta and to a lesser extent from the myometrium. PGF2alpha enhances the myometrial response to oxytocin and, after a latent period, stimulates contractions. The onset of parturition is normally associated with softening of the cervix, the mechanism of which is uncertain. Uterine contractions in the presence of a distensible cervix lead to parturition.


[Ossification of the auricle in partial pituitary failure with selective ACTH and STH deficiency (author's transl)]. An acquired partial pituitary insufficiency of unknown origin with selective ACTH and STH deficiency was demonstrated in a 44-year-old patient. The clinical course over many years corresponds to subclinical Addison's disease with occasional acute crises. Ossification of both auricular cartilages and anhidrosis were outstanding signs. There is possibly a connection between the glucocorticoid deficiency over many years with normal mineralocorticoids and the auricular cartilage ossification.


[Photosensitivity in children with malign tumors (author's transl)]. Photosensitivity (photoconvulsive reaction = PCR) has to be regarded as a genetically determined EEG-symptom, which is seen with markedly age dependent in up to 15% of healthy individuals. Correlations with epileptic and nonepileptic diseases (migraine, anorexia nervosa, psychovegetativ instability) are known. 157 children with malign tumors (Tab 1) were investigated. Children with Wilms' tumors showed significant more PCR than all other children with malign tumors (Tab. 2, 3). That finding may corroborate the hypothesis, that Wilms' tumor may be the most important clinical symptom of syndroms with congenital malformations (aniridy, hamartom, hemihypertrophy).


Release of cyclic AMP from the chicken thyroid stimulated with TSH in vitro. The thyroid lobes excised from one-day-old or 13-day-old chickens were incubated with or without TSH, and cAMP in the medium was determined. TSH induced the release of cAMP from thyroid lobes, resulting in the increase of cAMP concentration in the incubating medium. The release of cAMP into the incubating media in the presence of TSH from the thyroids of the chickens which were pretreated with TSH neonatally was more marked as compared to that from the thyroids of the control chickens. The light and electron microscopic examination of the thyroid lobes incubated with TSH showed that the morphological changes such as colloid droplets formation and enlargement of endoplasmic reticulum were induced.


Studies on reduction of lipolysis in adipose tissue on freezing and thawing. Adrenaline-induced lipolysis in fat cells was remarkably reduced when the cells were preincubated in a dry ice-aceton bath, but their adenylcyclase and lipase activities were not reduced. In the reconstructed lipid micelles which consisted of lipase-depleted lipid micelles and lipase-containing adipose tissue extract, adrenaline, theophylline and DBcAMP-induced lipolysis was not found when lipase-depleted lipid micelles were preincubated in a dry ice-aceton bath but was found when lipase was preincubated.


[Morphological classification of pituitary adenomas and its value for clinical diagnosis]. Pituitary adenomas should be classified not only by their tinctorial affinities but also by their degree of differentiation. Then useful correlation to the clinical data can be obtained; On this principle our own collection of 299 tumors was classified in undifferentiated acidophilic, highly differentiated acidophilic GH cell-, highly differentiated acidophilic prolactin cell-adenomas, in undifferentiated mucoid cell-, highly differentiated mucoid ACTH cell-, highly differentiated mucoid TSH cell-adenomas, in chromophobic adenomas of small cell type and of large cell type, and in oncocytic adenomas. 95% of the cases with acromegaly based on undifferentiated acidophilic or highly differentiated GH cell adenomas. All patients with hypothalamic-hypophyseal Cushing's syndrome or Nelson's syndrome had undifferentiated mucoid cell adenomas or highly differentiated ACTH cell adenomas. In cases with hyperprolactinemia prolactin cell adenomas or chromophobic adenomas of large cell type with ultrastructurally demonstrated very highly developed rough endoplasmic reticulum or endocrinologically inactive chromophobic adenomas of small cell type were found. In the latter cases the prolactin is probably produced not by the tumor but by the surrounding tumor-free pituitary tissue.


Human erythrocyte proteins associated with adenosine 3',5'-cyclic monophosphate action. Two adenosine 3',5'-cyclic monophosphate (cyclic-AMP)-binding protein factors (molecular weight 230,000) have been partially purified from human erythrocytes. One of these proteins seems to be different from the cyclic-AMP-binding component of the cyclic-AMP-dependent protein kinases. These protein factors are also capable of binding adenosine. We present data also on two forms of cyclic-AMP-dependent protein kinases (ATP: protein phosphotransferase, EC 2.7.1.37) partially purified from the cytosol of normal human erythrocytes. Kinase I has been classified as type I enzyme on the basis of its activation when preincubated with protamine, histone or NaCl. The substrate specificities of the two kinases and many of their kinetic parameters are rather similar. Their subunit structure is reminiscent of that of kinases obtained from other sources. The catalytic subunit of both enzymes reversibly cross-react with the regulatory subunit of kinase I from the rabbit red blood cell.


D-penicillamine and enzymatic activities. The influence of incubation with D-penicillamine on pure enzyme preparations and on enzymatic activities of serum and skin homogenates was investigated. Three of the nine enzymatic activities studied underwent significant changes. Such effects of D-penicillamine must be taken into consideration if therapeutic or unwanted actions of this drug are to be fully understood; they are elicited by concentrations reached under conditions used for human therapy.


The influence of exogenous and of membrane-bound phosphatidate concentration on the activity of CTP: phosphatidate cytidylyltransferase and phosphatidate phosphohydrolase. Rat liver microsomes were treated with phospholipase D to obtain microsomal membranes with varying amounts of membrane-bound phosphatidate. This treatment did not impair the activity of two microsomal-bound enzymes acting with phosphatidate as substrate, i.e. CTP: phosphatidate cytidylyltransferase and phosphatidate phosphohydrolase. The dependency of the activity of these enzymes on the concentration of membrane-bound phosphatidate was determined. Both enzymes showed a linear increase in activity with membrane-bound phosphatidate concentrations up to at least 100 nmol phosphatidate/mg microsomal protein. These results indicate that both enzymes have a large reserve capacity and suggest that the enzymes are operating intracellularly, i.e. at phosphatidate concentrations of 5-10 nmol/mg endoplasmic reticulum protein, far below their maximal capacity. The ratio of phosphatidate conversion into CDP-diglyceride and 1,2-diglyceride seems to be constant for a large range of membrane-bound phosphatidate concentrations. The membrane-bound enzymes cannot utilize phosphatidate substrate present in heat-denatured membranes, but are active on phosphatidate incorporated into membranes of phospholipid vesicles.


Investigation on the kinetic mechanism of octopine dehydrogenase. A regulatory behavior. The kinetic scheme of octopine dehydrogenase of Pecten maximus L., a monomeric enzyme obeying a bi-ter sequential mechanism, was completed, essentially in the forward reaction, by steady-state studies over a wide range of substrate concentration at pH 7.0. Deviation from the Michaelis-Menten behavior with respect to NAD+ and other significant kinetic data led us to ascribe for octopine dehydrogenase mechanism the mnemonical enzyme concept. In addition, another regulatory behavior can be envisaged involving the formation of two dead-end complexes enzyme.NADH.D-octopine and enzyme.NAD+.pyruvate.L-arginine.


A novel type of endotoxin structure present in Bordetella pertussis. Isolation of two different polysaccharides bound to lipid A. The endotoxin of Bordetella pertussis was cleaved by mild acidic hydrolysis to yield a polysaccharide (polysaccharide I, 15%), a glycolipid (63%) and lipid X (2%). Further treatment of the glycolipid with stronger acid released a second polysaccharide (polysaccharide II, 9%) and material similar to lipid A present in enterobacterial endotoxins. Both polysaccharides possess a single molecule of 3-deoxy-2-octulosonic acid as the reducing, terminal sugar. In polysaccharide II the octulosonic acid is phosphorylated in position 5 and presumably substituted in position 4; in polysaccharide I the octulosonic acid is not phosphorylated, but is substituted in position 5. Following treatment of the endotoxin with strong base, a fragment was isolated that contained bound, non-phosphorylated 3-deoxy-2-octulosonic acid, glucosamine phosphate and fatty acids. This indicated that polysaccharide I, like polysaccharide II, was bound to the lipid region of the endotoxin. The endotoxin structure thus defined is different from that proposed for the lipopolysaccharides of enterobacteria.


Kinetic evidence for interaction between aldolase and D-glyceraldehyde-3-phosphate dehydrogenase. The possibility of interaction between purified rabbit muscle aldolase and D-glyceraldehyde-3-phosphate dehydrogenase was studied by rapid kinetic methods, by analyzing the kinetics of the consecutive reaction catalyzed by the coupled enzyme system. The Km of the intermediary product, glyceraldehyde 3-phosphate, produced by aldolase was determined in the coupled reaction for glyceraldehyde-3-phosphate dehydrogenase. Its value corresponds to that of the aldehyde (active) form of glyceraldehyde 3-phosphate, although in the given conditions the aldehyde leads to diol interconversion is faster than the enzymic reaction catalyzed by glyceraldehyde-3-phosphate dehydrogenase. We suggest that above a certain concentration of the enzymes the glyceraldehyde 3-phosphate produced by aldolase gets direct access to glyceraldehyde-3-phosphate dehydrogenase without participating in the aldehyde leads to diol interconversion which otherwise would occur if the substrate were to mix with the bulk medium.


Nucleotide sequence of part of the simian virus 40 Hind-D restriction fragment. The presumed initiation region of the VP2 gene. The nucleotide sequence at the beginning of the restriction fragment Hind-D from Simian virus 40 DNA has been derived by partial chemical degradation of (5'-32P)-labeled restriction fragments followed by analysis on polyacrylamide gel according to Maxam and Gilbert [Proc. Natl Acad. Sci. U.S.A. 74, 560-564 (1977)]. The sequence reported here is 140 nucleotides long. It contains an ATG codon which presumably corresponds to the initiator codon of the VP2 protein. This codon is preceded by an untranslated region which shows several interesting features, such as an alternation of (dA+dT)-rich and (dG+dC)-rich blocks.


Nucleotide sequence of the simian virus 40 Hind-K restriction fragment. The restriction fragment Hind-K represents 4.2% of the genome of Simian virus 40 (SV40) and is located near the middle of the late region. Its nucleotide sequence is reported here. It was mainly established by analysis of transcription products, synthesized by means of Escherichia coli RNA polymerase and nucleoside triphosphates, one of which was (alpha-32P)-labeled. Strand assignment was possible by hybridization of asymmetric, labeled transcripts of total SV40 DNA to filter-bound Hind-K fragment. Further information and unambiguous confirmation of the sequence was obtained by the use of direct DNA-sequencing methods. For this purpose the fragment was labeled at the 5' ends by means of polynucleotide kinase and [gamma-32P]ATP and redigested with a suitable restriction enzyme. The separated products were then either partially digested with snake venom diesterase for analysis by the 'wandering spot' method or partially degraded with the base-specific reagents dimethylsulphate or hydrazine for direct sequence analysis on gel. The Hind-K sequence is 219 base pairs long. The message strand is particularly rich in adenosine (39%) and purines. The nucleotide sequence cna unambiguously be translated into an amino acid sequence and the N-terminal codon of the viral protein VP1 gene could be identified. The amino-terminal part of VP1 is rich in proline and lysine. The nucleotide sequence of Hind-K codes also for the carboxyl-terminal part of the viral protein VP2 and VP3 genes, which partly overlap the VP1 gene.


Lanthanum as a tool to study the role of phosphatidylinositol in the calcium transport in rat parotid glands upon cholinergic stimulation. We describe the effects of lanthanum on protein secretion, potassium efflux, calcium uptake and phosphatidylinositol turnover stimulated by cholinergic agonists in rat parotid glands. Carbachol increases in vitro calcium uptake, protein secretion and K+ efflux through muscarinic receptor; however it fails to stimulate protein discharge or K+ release in a incubation medium free of calcium. Lanthanum inhibits calcium uptake, protein secretion and K+ efflux induced by carbachol without impairing protein discharge stimulated by norepinephrine through the beta-adrenergic receptor. Norepinephrine, in the presence of calcium in the incubation medium, stimulates the K+ efflux through the alpha-adrenergic receptor: this effect is suppressed by lanthanum. These results emphasize the role of increased influx of calcium in the cellular phenomena controlled by muscarinic or alpha-adrenergic receptors. Carbachol increases phosphatidylinositol turnover in the absence of calcium in extracellular medium; indeed it is shown that carbachol increases the rate of phosphatidylinositol breakdown and that lanthanum impairs this cholinergic effects. From these data it is suggested that the interaction between cholinergic agonist and muscarinic receptor could induce a stimulation of 'phosphatidylinositol turnover' which could control the calcium influx according to the gradient through the plasmalemma membrane.


The role of cyclic AMP in temperature-dependent changes of contractile force and sensitivity ot isoprenaline and papaverine in guinea-pig atria. Right and left guinea-pig atria responded to decreasing temperatures (42-27 degrees C) with elevation for force of contraction and concomitant increases in cAMP. When atria were rapidly cooled from 42 to 27 degrees C the increase in cAMP occurred prior to the onset of the inotropic responses. Papaverine (3 X 10(-5) M) potentiated the effects of temperature on cAMP and force of contraction on left atria driven at 0.5 Hz. On right atria beating spontaneously at frequencies above 2 Hz papaverine only potentiated the effect of decreasing temperatures on the response of cAMP but not on that of force of contraction. Time course studies of the effects of isoprenaline (3 X 10(-8) M) on right atria at 27 degrees C showed large inotropic responses to isoprenaline which were accompanied by increases in cAMP. At 42 degrees C the responses of force of contraction and cAMP to isoprenaline occurred faster and were only short-lasting. As with the time courses for isoprenaline, dose-response curves for the effect of isoprenaline and papaverine on cAMP content and force of contraction also appeared to be shifted towards higher levels at hypothemia. However, pD2 values reflected increases in affinity for inotropic, but not for the cAMP responses to isoprenaline and papaverine at hypothermia. These results show that cyclic AMP is involved in the inotropic responses to hypothermia, but not in the supersensitivity of heart to isoprenaline and papaverine as observed at low temperatures.


Increases in plasma cyclic AMP dependent on endogenous catecholamines. Administration of tyramine (with or without phentolamine) as well as induction of ether anesthesia or insulin hypoglycemia caused a sharp increase in plasma cyclic AMP in rats. Based on the findings that the treatment of rats with reserpine, 6-hydroxydopamine, cocaine or propranolol totally abolished tyramine-induced increases in plasma cyclic AMP, it was concluded that catecholamines released from sympathetic neuronal terminals by tyramine could activate adenylate cyclase via the stimulation of postsynaptic beta-adrenoceptors. In contrast, catecholamines secreted from adrenal medulla were largely responsible for the increase in plasma cyclic AMP induced by ether anesthesia; whereas glucagon, in addition to adrenal catecholamines, played a significant role in hypoglycemia-induced increases in plasma cyclic AMP. Assay of plasma cyclic AMP following these stimuli is very promising as a test for adrenergic activities in experimental and clinical studies.


Correlation between isoprenaline-stimulated synthesis of cyclic AMP and occurrence of beta-adrenoreceptors in immature erythrocytes from rats. When increasing reticulocytosis (up to 80%) was induced in rats by treatment with acetyl-phenylhydrazine (up to 70 mg/kg on 3 consecutive days) synthesis of cAMP stimulated by isoprenaline in intact red blood cells and in the related membrane preparations increased up to 100-fold. At the same time the density of beta-adrenoreceptor sites (measured by ligand binding with (3H)(-)-dihydroalprenolol) in the related membrane preparations was only increased about 5-fold. There was however a linear correlation between the increase of isoprenaline-stimulated enzyme activity and the receptor density in membrane preparations. The results indicate that mature red cells contain beta-adrenoreceptor sites but little or not adenyl cyclase activity. Thus, during the maturation process of these cells, enzyme activity and receptor densities decrease at different speeds indicating a different turnover of these two entities of the cytoplasmic membrane.


Exchange of tritium from randomly tritiated taurocholate by microbial bile salt oxidoreductases. Tritium distribution on randomly labelled taurocholate (TC) was estimated at 28%, 4%, 1% and less than 0.5% on the hydrogens opposite the 3alpha-, 7alpha- and 12alpha-OH groups and taurine moiety respectively. Anomalously, C. perfringens 3alpha-hydroxysteroid dehydrogenase (3alpha-HSDH) catalyzed tritium loss of 36% on formation of 7alpha-, 12alpha-dihydroxy-3-keto-5beta-cholanoate, implying additional losses of tritium at other sites by this enzyme.


Inhibition of prostaglandin-induced cyclic AMP accumulation in the rat anterior pituitary by alrestatin. Alrestatin at 25-1 X 10(-4) M inhibited the accumulation of cyclic AMP induced by prostaglandin E2, but not theophylline, in the rat anterior pituitary in vitro. Somatostatin, at lower concentrations, inhibited both; maximal inhibition of the prostaglandin effect was greater with alrestatin. As cyclic AMP is considered to be a mediator in induced-hormonal release, it appears from the present findings that alrestatin may be of potential use in altering hormonal release.


Neuronal responses to extracellularly applied cyclic AMP:Role of the adenosine receptor. At low doses, theophylline blocks the neuronal depressant effects of 5'-AMP, but not cyclic AMP. Higher doses (100 mg/kg) block cyclic AMP responses and reduce the effects of noradrenaline and GABA. It is concluded that cyclic AMP does not depress neurones via the adenosine receptor.


A synergistic interaction between the teratogenic effect of trypan blue and dietary deficiency in the rat. There was an increased incidence, compared to controls, of exencephaly and microphthalmia in the offspring of rats fed a vitamin D deficient diet and injected with trypan blue on day 9 of gestation. Oral vitamin D did not reverse the effect.


Interactions between age and the neuroendocrine and immune systems. Three conclusions are suggested by some of the recent work on aging, immunology and the neuroendocrine system. 1) There appears to be sufficient data to implicate the neuroendocrine system in both the maturation and the senescence of at least some components of the immune system. 2) The thymus by its presence or its absence appears to influence certain functions of the pituitary; thus, there appears to be a possible reciprocal relationship between the pituitary and the thymus. 3) Changes in the levels of pituitary hormones or hormones that are controlled by the pituitary can restore in older rats and mice certain functions that are generally considered as part of the immune surveillance and defense system. Consequently, it can be hoped that further studies of neuroendocrine-immune relationships might lead to an understanding of some of the causes for the decline in immune competence with age in mammals.


The morphology of interneuronal synaptogenesis: a review. This review has endeavored to present current findings on the morphology of synaptogenesis, summarized in Fig. 5. Though much has been discovered about the order of events and the origin of organelles, the crucial questions of why these events occur at the moment in space and time remain largely unanswered. For an understanding of the molecular basis of neuronal recognition, and of the assembly and maintenance of a functional synaptic active zone, further extensive multidisciplinary studies will be necessary.


Interactions between neurons and their targets during in vivo synaptogenesis. In vivo synaptogenesis is described in a simple vertebrate system, the chick ciliary ganglion, a parasympathetic autonomic ganglion. An attempt is made to integrate anatomical, physiological and biochemical observations during synapse formation in the ganglion and in the peripheral target structures; the iris, ciliary muscle, and smooth muscle of the choroid coat. The relationship between synaptogenesis and neuron survival is explored, and it is shown that a critically timed interaction between the neuron and target organ is necessary for full neuronal maturation and survival. The existence of an active competition between neurons for survival is documented, and the possible relationship between neuronal cell death and specificity of connections is discussed.


Effect of two phosphodiesterase inhibitors, cyclic adenosine 3':5'-monophosphate, and a beta-blocking agent on human sperm motility. Mann-fructose fluid (MF), and MF plus caffeine, MF plus pentoxifylline, MF (dibutyryl cAMP), MF plus propranolol, and MF plus propranolol plus dibutyryl cAMP were individually added to aliquots of semen samples obtained from 18 normal men. These drugs were added to a final concentration of 0.6 mM. One aliquot with no addition served as control. Samples were incubated at 37 degrees C and observed by light microscopy at 30 minutes and at 1,2, and 4 hours after obtained the material. At each observation time, semen quality was evaluated by determinating the percentages of forwardly progresssive spermatozoa, slowly progressive spermatozoa, "in situ" motile spermatozoa, live and nonmotile spermatozoa, and dead spermatozoa. Mann-fructose fluid resulted in a decrease in motility and the duration of activity of spermatozoa, Caffeine seemed to neutralize the deleterus effect on the buffer, whereas pentoxifylline and cAMP seemed to increase the duration of activity of spermatozoa. Propranolol resulted in a dramatic decrease in motility, an effect that could not be neutralized by the simultaneous addition of cAMP.


[Interaction of catecholamines and corticosteroids during the process of muscle fatigue]. After prolonged (8 hrs) swimming, the level of corticosteroids was reduced and the contents of aldosterone and DOC in the blood was increased in the rats. At incubation of these rats' adrenals in the L-thyrosine substratum the synthesis of adrenaline was reduced. Addition of hydrocortisone, prednisolone and corticosterone into the medium as well as in vivo administration of these increased the adrenaline synthesis in swimming rats and did not alter it in intact rats. Neither in vitro addition of aldosterone nor in vivo administration of ACTH activated the process. The activating effect of glucocorticoids was absent at incubation of the adrenals with L-DOPA and L-noradrenaline. Synthesis of cathecholamines in the heart of swimming rats in the presence of L-thyrosine and L-DOPA as precnrsors was suppressed; neither was it restored by administration of corticosteroids both in vitro and in vivo. This suggests that one of the reasons for suppression of the catecholamine synthesis in adrenals at obvious physical fatigue is the decrease of the glucocorticoid activity.


Worldwide controversies in gestational trophoblastic neoplasms. This article reviews worldwide controversies concerning gestational trophoblastic neoplasms (hydatidiform mole, invasive mole, and choriocarcinoma). The epidemiology, endocrinology, histopathologic grading, classification, diagnosis, treatment, management and follow-up (including chemotherapy, irradiation, and immunotherapy) of gestational trophoblastic neoplasms - particularly molar pregnancies - are discussed; and ways to help create a standardized classification system and provide optimal treatment for each type of patient are suggested.


Lack of effect of bile in pathogenesis of secretion in mechanical small bowel obstruction. In dogs, the principal lesion in the ileal mucosa above a mechanical obstruction is the induction of a secretory state in which there is a net loss of water and electrolytes into the intestinal lumen. The role of stagnated bile as a possible agent provoking this secretion was explored by ligating the common bile duct and diverting the bile by means of an anastomosis between the gall bladder and the ileum distal to the obstruction. 7 days after creation of an obstruction with bile diversion, the functional and morphological changes in the obstructed loop were almost identical to those occurring in animals with obstructions without bile diversion. The secretory response above the obstruction continued unabated, with levels of cAMP in the secreting mucosae lower than in control tissues. These results suggest: (a) that the secretion in the obstructed intestine is not influenced by the presence of bile, and (b) that this secretion may not be secondary to an increase in mucosal cAMP levels.


Actions of salbutamol in late pregnancy: plasma cyclic AMP, insulin and C-peptide, carbohydrate and lipid metabolites in diabetic and non-diabetic women. Salbutamol was administered intravenously in dose increasing from 3.75 to 22.5 microgram/min to 5 non-diabetic and 7 diabetic women in the last trimester of pregnancy. In diabetic as well as non-diabetic women the diastolic blood pressure fell progressively with increasing doses, and the systolic BP and heart rate increased at doses above 7.5 microgram/min. The effect of fetal heart rate was less pronounced than the effect on maternal heart rate. Cyclic AMP levels in plasma were similar in non-diabetic and diabetic women before salbutamol. Twenty min following 3.75 microgram/min a significant increase was seen in both groups. The peak increase (3-5 fold) was higher in the diabetic than in the non-diabetic women. Plasma insulin and C-peptide levels rose in a dose-dependent manner in the non-diabetic and four of the diabetic women. However, in three of the diabetic women the insulin level was unaffected by salbutamol and C-peptide was almost undetectable. Plasma concentrations of glucose, glycerol, NEFA and 3-HB were higher in the diabetics than in the non-diabetics before salbutamol and the elevations induced by salbutamol were also significantly larger in the diabetic women. The present data show that salbutamol in doses employed clinically may cause pronounced metabolic effects, especially in diabetic women, and it is suggested that when intravenous infusion of salbutamol is given to pregnant diabetic women not only cardiovascular but also some metabolic variable such as glucose should be carefully monitored.


Hepatic insulin responsiveness in patients with endogenous hypertriglyceridaemia. Plasma insulin response to oral glucose, insulin resistance, and insulin suppression of hepatic glucose production were studied in 11 normal subjects and 11 hypertriglyceridaemic patients. Patients with hypertriglyceridaemia had a significantly higher insulin response to oral glucose. Insulin resistance was also significantly greater in hypertriglyceridaemic subjects as determined by measuring the steady-state plasma glucose response during a continuous infusion of epinephrine, propranolol, glucose, and exogenous insulin. Insulin suppression of hepatic glucose production was calculated from the results of two studies in which glucose turnover rate was measured by a continuous infusion of 3H-2-glucose. The first study was performed under conditions of basal insulin secretion, and the second carried out at steady state exogenous insulin levels of approximately 100 muU/ml. The results indicated that basal hepatic glucose production was the same in both groups, and was suppressed to an equal degree by physiological levels of insulin. These data demonstrate that hepatic glucose production can be suppressed to an equal degree in normal and hypertriglyceridaemic subjects at comparable circulating insulin levels, at the same time that resistance to insulin-stimulated glucose uptake is observed in the hypertriglyceridaemic individuals.


Epstein-Barr virus-determined nuclear antigen in malignant epithelial cells of nasopharyngeal carcinoma. Tumor tissue from lymph node metastasis of nasopharyngeal carcinoma (NPC) was successfully heterotransplanted into an athymic nude mouse and the tumor grown in the nude mouse was identical in its morphology to that form the patient by optical and electron microscopy. Tumor cells at passage 2 were dispersed in vitro by enzymic digestion and smeared. Epstein-Barr virus-determined nuclear antigen (EBNA) was demonstrated in malignant epithelial cells of the smear.


Antitumor activity of a new amino acid derivative, N6,N9-bis-(butyloxycarbonylaminomethyl)-L-citrulline. Approximately 800 amino acid derivatives have been synthesized and screened in order to evaluate their antitumor activity against various transplantable rat ascites hepatomas. Among them, N6,N9-bis(butyloxycarbonylaminomethyl)-L-citrulline (A-924) was found to be highly effective against various rat ascites hepatomas. A-924, when given orally, exhibited prolongation of survival of rats implanted intravenously with ascites hepatoma cells such as AH-44, AH-66, AH-130, AH-66F, or AH-41C.


X-linked and autosomal genes controlling mouse alpha-galactosidase expression. Analysis of F2 and backcross animals has confirmed the X-chromosome linkage of Ags, the structural locus for mouse alpha-galactosidase. The position of Ags has been located in the X chromosome, 9 centimorgans from Mo, and the gene order is centromere-Hq-Bn-Ta-Mo-Ags. A variation in the developmental expression of alpha-galactosidase activity, inherited as an autosomal trait, has been characterized using recombinant inbred lines of mice. Among certain recombinant inbred lines, the variation appears to segregate as a single major locus.


Heterozygous effects on fitness of EMS-treated chromosomes in Drosophila melanogaster. The heterozygous effects on fitness of second chromosomes carrying mutants induced with different doses of EMS were ascertained by monitoring changes in chromosome frequencies over time. These changes were observed in populations in which the treated chromosomes, as well as untreated competitors, remained heterozygous in males generation after generation. This situation was achieved by using a translocation which links the second chromosome to the X chromosome; however, only untranslocated second chromosomes were mutagenized. Chromosomes were classified according to their effects on viability in homozygous condition. A preliminary homozygosis identified completely lethal chromosomes; secondary tests distinguished between drastic (viability index < 0.1) and nondrastic chromosomes. Chromosomes that were nondrastic after treatment were found to reduce the fitness of their heterozygous carriers by 3-5%. The data show that flies homozygous for these chromosomes were about 2.7% less viable per treatment with 1 mm EMS than flies homozygous for untreated chromosomes. By comparing the fitness-depressing effects of nondrastic EMS-induced mutants in heterozygous condition with the corresponding viability-depressing effects measured by Temin, it is apparent that the total fitness effects are several times larger than the viability effects alone. Completely lethal chromosomes derived from the most heavily treated material reduced fitness by 11% in heterozygous condition; approximately half of this reduction was due to the lethal mutations themselves.


[Operon study of riboflavin biosynthesis in Bacillus subtilis. XII. The determination of the ATP:riboflavin-5'-phosphotransferase and riboflavinsynthetase content in the cells with varying genotypes]. Activities of riboflavinkinase and riboflavinsynthetase were measured in 15 strains of Bacillus subtilis with different genotype. The increased level of riboflavinkinase was observed in strains, resistant to lumiflavin or lumichrome. Specific activity of riboflavinkinase was found to be about 100 times lower than that of riboflavinsynthetase. The regulation of biosynthesis of these enzymes seems to proceed non-coordinately. This phenomenon can be the sequence of the existence of many operators, controlling the flavinogenesis in Bac. subtilis.


[Genetic control over 7 Lpm-system allotypes of very high-density serum alpha2-lipoprotein the mink]. Population and hybridological analyses have demonstrated that the alloantigenic markers of very high density alpha2-lipoprotein of mink serum, Lpm7 and Lpm8, along with Lpm1, Lpm2, Lpm3, Lpm4, Lpm5, belong to a common immunogenetic system. 21 Lpm-phenotypes are determined at least by 36 genotypes; each of the 11 phenotypes is conditioned by a single genotype, each of the remaining 10 are conditioned by 2, 3 or 4 genotypes. The Lpm allotypes and allogroups are coded by the genetic units Lpm8, Lpm4, Lpm4,8, Lpm4,7, Lpm3,4,8, Lpm1,8, Lpm1,2,7, Lpm2,4,5,7, which behave as alleles. The last, six seem to be haploid sets of closely linked genes.


[Spectral studies of drug interactions with hemoglobin and cytochrome c (author's transl)]. We studied the drug-induced spectra of hemoglobin and cytochrome c using aniline, aminopyrine, hydrocortisone and prednisolone. The difference spectrum of methemoglobin induced by aniline or aminopyrine was similar to type II, and that of oxidized cytochrome c induced by aniline was similar to type I. Characteristic spectral change occurred with addition of hydrocortisone or prednisolone to each of methemoglobin, oxyhemoglobin and carboxyhemoglobin, and by addition of each drug to deoxyhemoglobin, respectively. Other spectral changes were attributed to oxidation-reduction reactions between the drugs and the hemoproteins. Each of the reduced forms of hemoglobin showed a higher affinity to aniline and aminopyrine than did methemoglobin. Each of the reduced forms of hemoglobin or reduced cytochrome c showed a higher affinity to aminopyrine than to aniline. It is interesting that not only cytochrome P-450, a hydrophobic hemoprotein, but also hemoglobin and cytochrome c, hydrophilic hemoproteins, showed the drug-induced difference spectra. These results suggest direct interactions between drugs and hemoproteins. Oxyhemoglobin had a lower affinity to drugs in hemolysate than in the buffer only.


[Hepatology and immunology].

[Hepatology and immunology]. Immunologic diseases of the liver are exogenous mostly initiated by virus or endogenous initiated by autoaggression. All virus-induced kinds of hepatitis are due to an immune response against inocculated hepatocytes. Therefore the hepatitis is limited to the period of complete elimination of virus-infected cells. A strong immune response therefore corresponds with an acute and short hepatitis whilest a weak immune response develops a chronic hepatitis. In contrast, autoimmune hepatitis based on a disorder of the immune system with some genetic background is always unlimited. Each cirrhosis developing from immunologic hepatitis is also an immunologic disease; a special variant is the autoimmune primary biliary cirrhosis. All in all, the number of immunologic liver diseases surmounts the remaining liver diseases due to intoxication of metabolic disorders.


[Lesions of the peripheral nerves in Wilson's disease. Electrodiagnostic findings (author's transl)]. Electrodiagnostical studies were carried out on three patients with Wilson's disease. The motor nerve conduction velocity was low but not markedly decreased. Some values of the sensory conduction velocity were decreased; the sensory action potentials showed low amplitudes and contained large amounts of late phases. Also, the electromyogram of one female patient showed denervation in the begin of treatment. There are no indications of polyneuropathy in Wilson's disease in the literature. There have been histological reports, however, which indicated lesions of the peripheral nerves in Wilson's disease as our own electrodiagnostical findings do.


Histologic classification of pure primary epithelial breast cancer. Current classifications of breast cancer are based on either morphologic descriptive or histogenetic criteria. The positive features and shortcomings of the most commonly used systems are presented. Our knowledge of breast cancer leads validity to a histogenetic topographic classification, a modification of which is suggested.


Ultrastructure of malignant histiocytoma arising in the acromion. The ultrastructural features of a malignant histiocytoma of the acromial process of the scapula were studied. Material was obtained from two surgical biopsy specimens and an amputation specimen from the tumor. Cells possessing characteristics of histiocytes, fibroblasts, xanthoma cells, and multinucleated giant cells were present throughout the tumor. Smaller numbers of undifferentiated cells  and lymphocytes were also observed. Intimate cytoplasmic interdigitations between adjacent tumor cells were found, and instances of degenerating intracytoplasmic cells, possibly representing phagocytosis, were observed. Specimens stained with periodic acid-Schiff reagent with and without exposure to diastase, examined by light microscopy, showed that numerous cells contained phagocytized material consisting of degenerating cells rather than cytoplasmic glycogen. Intraumor lymphocytes apparently represented an inflammatory reaction to the tumor. The tumor giant cells and xanthoma cells were probably modified histiocytes. Results of the study were compared with previous reports of ultrastructural studies of malignant histiocytoma of soft tissues. Fundamental similarities between such studies and this one suggested that the progenitor cell is a histiocyte, whether arising in bone or in soft tissues, and that the progenitor cell is capable of differentiation in both histiocytic and fibroblastic directions.


Pharmacological control of the hormonally modulated immune response. II. Blockade of antibody production by a combination of drugs acting on neuroendocrine functions. Its prevention by gonadotropins and corticotrophin. Injection of a combination of three drugs, 5-hydroxytryptophan, the alpha-blocker phentolamine and the neuroleptic drug haloperidol into mice before or together with sheep red blood cells (SRBC) induces a complete and long-lasting inhibition of antibody production to SRBC and leads to specific unresponsiveness. The mice unresponsive to SRBC respond normally to another antigen. Treatment with a combination of luteotropic (LH), follicle stimulating (FSH) and corticotropic hormone (ACTH) before administration of drugs and antigen prevents the immune blockade. Injection of SRBC induces an early elevation of LH in blood. This effect is prevented by previous administration of the three drugs in combination. The hormonal response to a second injection of the same antigen of mice previously made 'unresponsive' is different from that of immunized animals. The suppression of these hormonal changes which follow antigen injection by drugs acting on neuroendocrine regulation and cell membrane adrenergic receptors represents a step forward in efforts aimed at a pharmacological control of acquired immunity.


Myeloperoxidase-Cl--H2O2 bactericidal system: effect of bacterial membrane structure and growth conditions. The human-myeloperoxidase-Cl--H2O2 bactericidal system killed (i) smooth Enterobacteria spp. greater than or equal to rough and (ii) static-grown, stationary-phase bacteria greater than or equal to aerated-grown, log-phase. This is in contrast to human neutrophil granule extracts (involved in nonoxidative bactericidal mechanisms) that kill rough and aerated log-phase Enterobacteria spp. much more efficiently.


Intestinal enzyme activities in germfree, conventional, and gnotobiotic rats associated with indigenous microorganisms. Various enzyme activities involved in the active transport system, glycolysis, and digestion were assayed in various parts of the gastrointestinal tracts of germfree, conventional, and gnotobiotic rats associated with indigenous bacteria. The activity levels of alkaline phosphatase, glucose 6-phosphatase, adenosine triphosphatase, and disaccharidases in the upper small intestine were highest in all parts of the gastrointestinal tracts of various kinds of gnotobiotic, conventional, and germfree rats. Alkaline phosphatase, glucose 6-phosphatase, and adenosine triphosphatase activities in the upper small intestine of germfree rats were, respectively, 2.3-, 2.9-, and 1.7-fold higher than those in conventional rats. Similar to the results of these enzymes, sucrase, maltase, trehalase, and lactase activities in the upper small intestine of germfree rats were, respectively, 1.6-, 1.5-, 2.3-, and 1.8-fold higher than those in conventional rats. In various gnotobiotic rats, enzyme activity levels were intermediate between those in germfree and conventional rats. These findings suggest that those enzymatic activities are strongly depressed by the association with the indigenous microorganisms in the epithelial mucosa of the upper small intestine of rats. The levels of pyruvate kinase, hexokinase, and lactate dehydrogenase activities were highest, respectively, in the stomach, cecum, and the upper small intestine and cecum in all parts of the gastrointestinal tracts in various kinds of gnotobiotic, conventional, and germfree rats. It was also shown that six kinds of gastrointestinal bacteria, including lactobacilli, significantly depressed the enzyme activity levels to levels between those of the germfree and conventional rats in the upper small intestine of gnotobiotic rats.


Role of activated macrophages in resistance of congenitally athymic nude mice to hepatitis induced by herpes simplex virus type 2. Congenitally athymic nude (nu/nu) mice of a BALB/c genetic background were found considerably more resistant to the induction of focal necrotic hepatitis by herpes simplex virus type 2 (HSV-2) tha, were phenotypically normal littermates (nu/+) or BALB/c mice. The augmented resistance was age dependent, as it was only manifested in mice from 4 to 5 weeks of age. Studies of the course of infection showed that nude mice were able to restrain virus multiplication in the liver far better than normal mice in the early phase of infection. However, they seemed inferior to normal mice in eliminating the infectious process. In vitro investigation of peritoneal macrophages revealed that macrophages from 6-week-old nude mice exhibited accelerated spreading and were three times as restrictive in the replication of HSV-2 as macrophages from normal mice. However, no difference was found in the efficiency of adsorption/phagocytosis between  macrophages from nude and normal mice. The increased resistance of nude mice could be abolished by blockade of the microphage function of the mice by silica. Nude mice reconstituted at birth with thymus cells were just as susceptible to infection as normal mice. These data suggest that the increased resistance of nude mice to HSV-2 hepatitis is due to the presence of nonspecifically activated macrophages before infection.


Enhanced resistance against Junin virus infection induced by Corynebacterium parvum. The effects of intraperitoneal administration of Corynebacterium parvum on the course of Junin virus infection in mice were investigated. This treatment produced enhanced resistance to the virus infection, evidenced by an increase in both survival times and the proportion of survivors. The protective effect was dependent upon the dose of C. parvum, and 280 mug/g of body weight was found to be the optimal dose. In various experiments, about 80% of the infected animals receiving this dose survived, whereas survival ranged between 0 and 20% among untreated infected mice. Maximal protection was afforded by C. parvum when administered simultaneously with the virus. A smaller but significant degree of resistance was induced by C. parvum given 3 or 6 days after infection. C. parvum injected before infection was ineffective. Viral titers measured in the brains of C. parvum-treated and untreated mice at various times after infection were found to be comparable. In addition, there were no significant differences between circulating-antibody titers measured either by neutralization tests or by complement fixation. Depression of the reticuloendothelial system by treatment with silica particles also resulted in enhanced resistance to Junin virus infection, suggesting that the protective effect of C. parvum is not likely to be due merely to its capacity to stimulate macrophages. The present data, highlighting that the presence of high titers of Junin virus and disease do not necessarily correlated, suggest that in mice this disease is not the consequence of cell damage caused directly by the virus but of a still undefined indirect mechanism induced by the virus, not necessarily mediated by macrophages.


Production of interferon and serum hyporeactivity factor in mice infected with murine cytomegalovirus. Mice injected intraperitoneally with murine cytomegalovirus produced as many as 1,000 U of serum interferon. The response appeared biphasic, with maximum titers in the first phase detectable from 2 through 4 days after infection. A second phase peaked 10 days after infection. By carboxyhexyl-Sepharose affinity chromatography, the serum interferon behaved like lymphocyte interferon. The infected mice also produced substantial quantities of serum hyporeactivity factor (D.A. Stringfellow, E.R. Kern, D.K. Kelsey, and L.A. Glasgow, J. Infect. Dis. 135:540-551, 1977), although always in the presence of interferon. This factor was separated from the serum interferon by concanavalin A-Sepharose affinity chromatography.


Heterophil antigen in bovine sera detectable by immune adherence hemagglutination with infectious mononucleosis sera. Bovine sera for tissue culture use were shown to contain heterophil antigen of the Paul-Bunnell-Davidsohn type by immune adherence hemagglutination tests with sera from patients with infectious mononucleosis. The antibody titers observed were comparable to those determined by two other methods; i.e., the immune adherence hemagglutination test was found to be as specific as the differential absorption test with horse erythrocytes or the ox cell hemolysis assay,but it appeared to be the most sensitive of the three procedures. All 16 individual or pooled bovine serum samples showed Paul-Bunnell-Davidsohn antigen at concentrations varying over a fourfold range. The potential usefulness of the immune adherence hemagglutination test for various Paul-Bunnell-Davidsohn antigen-related problems and implications of the observations are discussed.


Oral--facial pain: old puzzles, new postulates. Dental examples are given (e.g. referred pain, V neuralgia) which exemplify the multifactorial and puzzling nature of pain. Pain results from a noxious stimulus activating some of the small-diameter myelinated and unmyelinated afferent nerve fibres that innervate skin, mucosa, teeth etc. It is complex experience including the sensation experienced as well as the emotional, cognitive and motivational reactions evoked by the stimulus. Pain is not a simple function of the magnitude of damage caused by the stimulus, but also depends on factors such as the person's emotions, past experience of pain, and other concomitant sensory experiences (e.g. with acupuncture). Early postulates of pain failed to take all these factors into account. In the V system (Fig. 1), the specificity theory would maintain that there exists a 'private pain path' from oral-facial tissues to the cerebral cortex, with nerve fibres and brain cells in this path responsive only to stimulu of a noxious character (Fig. 2). But anatomical, physiological and psychological observations have failed to give full support to this theory and other postulates of pain. Although some details of the more recently proposed gate control theory have not been sustantiated, this theory has provided a good general framework for viewing pain and has stimulated much recent research. It emphasizes sensory interaction between large and small-diameter afferent nerve fibres in the gate control system, and regulation over central pain transmission through the gate by descending central controls (Fig. 3). In the V system, brain cells involved in central pain transmission have now been found (Fig. 4), and sensory interactions and descending controls on these brain cells noted (Fig. 5). Neural mechanisms such as these have been implicated in pain, and in its control by procedures such as acupuncture, suggestion, distraction and narcotic analgesia.


Persistent and boosterable IgE antibody production in mice injected with low doses of ovalbumin and silica. The results of the present experiments show that Aerosil, an amorphous submicroscopic silica, is able to stimulate in Swiss outbred mice the production of IgE antibodies to a single 1 microgram dose of Ovalbumin (OA) in experimental conditions in which detectable IgG1 antibodies were not produced. The production of IgE anti-OA antibodies stimulated by silica was persistent for a least 4 months, and demonstrated a secondary stimulation. Neither IgE nor IgG1 anti-OA antibodies were demonstrable when mice were immunized with OA alone or OA with aluminum hydroxide.


Coordination of institution and parole services: an innovation within California's Civil Addict Program. Parole outcome was measured 1 year after release of 397 narcotic addicts processed by an experimental program in which both institution and parole services were administered together. Conparison of outcome of this group to the outcome of 361 traditionally processed addicts revealed a positive effect on parole outcome of violators (those with prior parole experience) compared to new commitments (p less than .05). This was particularly true for the community parole unit which made most use of community resources (p less than .05). Community adjustment of some narcotic addicts may be increased by administrative interdependence and by optimal use of other community resources. The future for such programs seems brightest at the local level of government.