Tumor antigen induction by the envelope-defective Bryan strain of Rous sarcoma viruses. Chicken, quail, and turkey cells were infected and/or transformed by various avian leukosis or sarcoma oncovirus (ALSV) strains as well as by the envelope-defective Bryan high-titer strain of Rous sarcoma virus [BH-RSV(-)]. All fibroblast-transforming avian sarcoma viruses (ASV), including BH-RSV(-), were able to induce the expression of the previously described avian tumor-specific cell-surface antigen(s) (TSSA). Thus TSSA was group-specific for all tested transforming ASV. Since BH-RSV(-) is a stable deletion mutant lacking the capacity to code for the 85 000d major virus envelope glycoprotein (gp85), it seemed unlikely that the group-specific antigen determinants of gp85 that were recently detected on ALSV-infected cells could account for TSSA expression.


Restriction to the cell nucleus of virus-induced host RNA's in a rat kidney tumor induced by polyoma virus. RNA transcripts of repetitive DNA of WF rat kidney, which were derepressed during transformation by polyoma virus, were not transported to the cytoplasm in the tumor cells. Transcripts of repetitive DNA found in the normal cell nucleus were all transported to the cytoplasm in the tumor cells, including those restricted to the nuclei in the normal cells.


Pathology of hepatomas and other liver abnormalities in English sole (Parophrys vetulus) from the Duwamish River estuary, Seattle, Washington. Liver abnormalities were found, by gross and histopathologic examination, in 92% of the English sole (Parophrys vetulus) from the Duwamish River Estuary, Seattle, Washington. Hepatomas were found in 32% (20 of 62) of the English sole. Other observed liver aberrations included increased fatty vacuolation, congestion, structure disarray, increased size and number of melanin-macrophage centers, centrolobular fatty degeneration and necrosis, increased amounts of perivascular connective tissue, intercellular melanin deposits, and hepatocellular hypertrophy often associated with the presence of bizarre nuclei and/or multiple nucleoli. Livers evidencing microscopic lesions were usually discolored. Livers containing hepatomas were often mottled yellow or tan and brown; occasionally, hepatomas were visible as tan or white nodules. Although the cause(s) of the liver abnormalities has not been conclusively identified, chemical analyses of Duwamish River English sole have detected polychlorinated biphenyl (PCB) levels of about 1.5 ppm (dry wt) in total body tissue. Many of the above-mentioned abnormalities, with the exception of hepatomas, have been observed in fish exposed to PCB's.


Effect of a mouse mammary tumor virus-derived protein vaccine on primary tumor development in mice. The vaccines used in this study were derived from purified murine mammary tumor virus (MuMTV) preparations. Approximately 60% of the protein fractions consisted of the major viral membrane glycoprotein gp52. Inoculation sc of 10 microgram MuMTV-S-derived vaccine significantly delayed the appearance of primary mammary tumors in GR and BALB/cfC3H mice (strains with high incidences of mammary cancer); in BALB/c and C3Hf mice, which have a moderate tumor incidence at an advanced age, this treatment resulted in a slight and substantial acceleration, respectively, of primary tumor development. The induced cellular immune reactivity for vaccination, as measured with the in vivo Winn test and the in vitro leukocyte adherence inhibition assay, was strongest in the GR strain as compared to the BALB/c strain. The titer of antibodies to tumor cells, as estimated by membrane immunofluorescence, was also higher in the GR strain. In BALB/cfC3H mice, the influence of different vaccination schemes with an MuMTV-O-derived protein vaccine on primary tumor development was studied. Before sc injection, the vaccine was precipitated on alum. A dose of 10 microgram vaccine resulted in a 61% decrease in tumor incidence. Two or five additional booster injections with 1 microgram protein vaccine had no beneficial effect, although the amount of antibody measured was increased after boosting.


Genetic control of immune responses to Moloney leukemia virus in rats. When BN and LEW rats were immunized with untreated or with inactivated Moloney murine leukemia virus (M-MuLV), BN rats produced high antibody responses to the p15, p30, and gp70 antigens of the virus, whereas LEW rats were low responders to these antigens. BN rats also exhibited a high response and LEW rats a low response when the two strains were immunized with purified p30. Studies of (LEW X BN)F1 and backcross rats suggested that factors associated with AgB exerted major influences on responses to antigens of M-MuLV and that other factors were also important. When other rat strains representing 5 AgB alleles were tested, some were high and some were low responders to M-MuLV, and responses to p15, p30, and gp70 were not always parallel. Since M-MuLV replication was greater in cells of BN rats than in cells of LEW rats, replication of M-MuLV may have influenced the levels of responses to some viral antigens. Control of virus replication appeared to be due to cell mechanisms rather than to the environment of the host.


Leukemogenicity of clonal isolates of murine leukemia viruses. The leukemogenicity of three types of cloned, in vitro grown murine retroviruses was studied. Two Moloney virus clones caused leukemia, as did five clones of the B-tropic endogenous virus of BALB/c mice. Neither of two clones of N-tropic BALB/c virus caused leukemia in Fv-1n/n mice, and the viruses were not recoverable from the animals. The ability to induce leukemia therefore appeared to reside in the genome of at least certain nondefective murine retroviruses.


Lymphocytotoxic autoantibodies eluted from in vivo propagating sarcoma cells of mice. Eluates of SEYF-a cells (from polyoma virus-induced sarcoma grown in A.BY mice) containing cytotoxic anti-SEYF-a antibodies mediated complement-dependent cytotoxicity of normal lymph node cells. Some lymphocytotoxicity was also mediated by sera of tumor-bearing mice, but the pattern of cytotoxicity was different from that exhibited by tumor eluates. The titer of lymphocytotoxic antibodies in the serum was lower than the titer of anti-SEYF-a antibodies, whereas eluates exhibited similar titers toward both target cells. These results indicated that tumor cells selectively absorbed the lymphocytotoxic antibodies from the serum. Tumor eluates had cytotoxic activity also against thymocytes, splenocytes, and even bone marrow cells. The strain distribution of sensitive thymocytes to the antibodies present in eluates indicated that Thy 1 was not involved. Unfractionated eluates as well as IgG fractions isolated from eluates were active.


Observations on hepato-cellular carcinoma in the Sudan. The clinical and histopathological features of hepato-cellular carcinoma in 24 Sudanese patients seen in Khartoum Civil Hospital in a 4 year period are described. The tumour represented 1.97 per cent of total medical admissions and constituted 40 per cent of all malignancies seen. The age range was 30 to 88 with a mean age of 54 years. The male:female ratio was 6:1. A regional variation was apparent and is possible that the disease is more common in the Western Sudan than in other parts of the country.


Recovery nystagmus.

Recovery nystagmus. Secondary nystagmus is frequently seen following cessation of prolonged unidirectional vestibular stimulation. It is explained on the basis of an adaptation during the application of the stimulus which leads to an apparent stimulus in the opposite direction when the stimulus is removed. The same phenomenon would be expected with vestibular disease when after a period of adaptation to the vestibular asymmetry, the affected ear recovers some or all of its function. The seondary nystagmus in this instance beats toward the affected ear and has been termed recovery nystagmus because it is generated by recovery of function. Two cases with recovery nystagmus following acute attacks of vertigo are presented.


Herpes simplex virus and human cytomegalovirus replication in WI-38 cells. III. Cytochemical localization of lysosomal enzymes in infected cells. Cytochemical localization of the lysosomal enzymes acid phosphatase and arylsulfatase in cells infected by herpes simplex virus (HSV) or human cytomegalovirus (CMV) showed the following interactions between viruses and host cell lysosomes: (i) many enveloped progeny viruses were located within cytoplasmic vacuoles containing lysosomal enzyme activity; (ii) naked cytoplasmic capsids appeared to acquire an envelope by budding directly into lysosomes; and (iii) many of the cytoplasmic dense bodies that are characteristic of CMV-infected cells and are thought to represent noninfectious aggregates of CMV structural proteins (I. Sarov and I. Abady, Virology 66:464-473, 1975) also acquired a limiting membrane by budding into lysosomes. Autophagy of other cytoplasmic elements was not observed, suggesting that there is some specificity involved in the association of viral particles and CMV dense bodies with lysosomes. Despite the presence of potentially destructive hydrolases, there was little evidence of significant morphological damage to intralysosomal viruses, and high titers of infectious particles were released into the medium. It would therefore appear that significant levels of HSV and CMV infectivity normally persist even though many progeny particles are directly exposed to lysosomal enzymes.


FBJ osteosarcoma virus in tissue culture. III. Isolation and characterization of non-virus-producing FBJ-transformed cells. Hamster and rat cell lines have been established that have been transformed by FBJ murine sarcoma virus (FBJ-MuSV) but that do not produce virus. The hamster cell line originated from an osteosarcoma that appeared in a hamster inoculated at birth with an extract of a CFNo1 mouse FBJ-osteosarcoma. The rat cell line was obtained by transferring the FBJ-MuSV genome to normal rat kidney cells in the absence of the FBJ type C virus (FBJ-MuLV), which, usually in high concentration, accompanies the FBJ-MuSV. Both transformed hamster and rat cell lines contain the FBJ-MuSV genome, which can be rescued by ecotropic and xenotropic murine type C viruses. This rescued genome produces characteristic FBJ-MuSV foci in tissue culture and, in appropriate animal hosts, induces osteosarcomas typical of those induced by FBJ-MuSV. FBJ-MuSV was isolated originally from a parosteal osteosarcoma that occurred naturally in a mouse. Since there was no previous history of passage of the agent through any other animal species, these non-virus-producing hamster and rat cells transformed by FBJ-MuSV should be very helpful in molecular studies examining the origin of spontaneous sarcoma genomes in mice.


Assembly of viral membranes: nature of the association of vesicular stomatitis virus proteins to membranes. To explore the interaction of vesicular stomatitis virus (VSV) proteins with cellular membranes, we have isolated membranes from infected cells that have been radioactively pulse-labeled. We have found conditions of isolation that result in membrane preparation which contain primarily the VSV membrane protein (M) and glycoprotein (G). Both of these proteins are very firmly attached to membranes: conditions known to release peripherally associated membrane proteins from membranes (S. Razin, Biochim, Biophys. Acta 265:241-246, 1972; S. J. Singer, Annu. Rev. Biochem. 43:805-826, 1974; S. J. Singer and G. L. Nicholson,  Science 175:720-731, 1972) are ineffective in detaching either the G or the M protein. The results of trypsin digestion of these membrane fractions suggest that the M protein resides primarily on one side, the cytoplasmic side of cellular membranes, whereas the glycoprotein has been transported to the lumen of the membrane vesicle. However, we present evidence that the glycoprotein is transmembranal and that approximately 3,000 daltons of one end of the molecule is on the cytoplasmic side of the membrane. We have also found that undenatured VSV M protein contains  a trypsin-resistant core with a molecular weight of 22,000. This region of the M protein is trypsin-resistant regardless of its association with membranes.


Effects of 2'-deoxy-2'-azidocytidine on polyoma virus DNA replication: evidence for rolling circle-type mechanism. Rolling circle-type molecules were found in polyoma virus-infected cells after inhibition of DNA synthesis with 2'-deoxy-2'-azidocytidine. The circular DNA molecules were always relaxed and of polyoma length. Most of the attached tails were less than two times the length of the polyoma genome, but tails with a length of up to 4.75 times the genome were also found. After cleavage of the total pool of replicating molecules with either endo R.EcoRI or endo R.BamI, Y-shaped molecules with replicated portions of various lengths were generated from rolling circle-type molecules. Moreover, after cleavage, Y-shaped molecules with three unequal arms were found, which could be explained as derived from the tail in rolling circle-type molecules starting from the normal origin, i.e., 29% from the endo R.EcoRI cleavage site. Rolling circle-type molecules were also found during a normal, noninhibited infection cycle. In such cells, a relatively higher frequency of rolling circle-type molecules was observed late during infection. Compared with control cultures, cultures inhibited with 2'-deoxy-2'-azidocytidine showed a greater amount of rolling circle-type molecules relative to normal replicative intermediates. 2'-Deoxy-2'-azidocytidine has previously been shown to inhibit the initiation of new rounds of replication; thus, the result obtained here indicates that a rolling circle-type mechanism is independent of the reinitiation of DNA synthesis.


T1 oligonucleotide maps of N-, B-, and B leads to NB-tropic murine leukemia viruses derived from BALB/c. We previously described and characterized RNase T1 RNA fingerprints of an N-, a B-, and five B leads to NB-tropic murine leukemia viruses derived from BALB/c mice (Faller and Hopkins, J. Virol. 23:188-195, 1977, and J. Virol. 24:609-617, 1977). These viruses share the majority of their large RNase T1-resistant oligonucleotides, but each possesses some "unique" oligonucleotides relative to the others. We have ordered the large T1-resistant oligonucleotides of the N-, the B-, and one NB-tropic virus relative to the 3' end of their genomes to obtain oligonucleotide maps. These maps indicate that (i) the large T1 oligonucleotides shared by the N-, B-, and NB-tropic viruses probably occupy the same relative positions on their genomes; (ii) the 14 T1 oligonucleotides that differ between the N- and B-tropic viruses are derived from regions scattered along the genomes;  and (iii) an oligonucleotide that is present in five NB-tropic viruses but not in their B-tropic virus progenitors lies toward the 5' end of the NB-tropic virus oligonucleotide map.


T1 oligonucleotides that segregate with tropism and with properties of gp70 in recombinants between N- and B-tropic murine leukemia viruses. We have analyzed large RNase T1-resistant oligonucleotides derived from the genomes of 16 recombinants between N- and B-tropic murine leukemia viruses of BALB/c. The parental viruses, designated SP-N and LP-B, differ in several phenotypic or biochemically defined properties: N- or B-tropism; XC plaque morphology, electrophoretic mobility of three virion proteins (p15, p30, and gp70); ability to induce GIX antigen on infected cells; presence of 6 to 8 (out of 36 to 38 analyzable) large T1 oligonucleotides. One SP-N-specific T1 oligonucleotide was inherited by all 16 N-tropic recombinants and, thus, appears to be linked to N-tropism. This oligonucleotide lies in the 5' third of the oligonucleotide map of SP-N. One LP-B-specific T1 oligonucleotide was inherited by all 11 recombinants whose gp70  has an electrophoretic mobility like that of LP-B gp70 and that, like LP-B, fail to induce GIX antigen. This oligonucleotide lies in the 3' third of the oligonucleotide map of LP-B.


Bovine and ovine leukemia viruses. I. Characterization of viral antigens. A comparative study on several virus-specific antigens of bovine and ovine leukemia viruses is presented. In addition to the major immunologically reactive, nonglycosylated antigen p21, which has a molecular weight of 21,000, both viruses share two glycoproteins of apparent molecular weights of 60,000 (gp60) and 32,000 (gp32), respectively. Immunological cross-reaction suggests that ovine and bovine leukemia viruses are genetically highly related.


In vitro RNA transcription by the New Jersey serotype of vesicular stomatitis virus. I. Characterization of the mRNA species. The in vitro RNA synthesis by the virion-associated RNA polymerase of vesicular stomatitis virus (VSV), New Jersey serotype, was compared with that of the serologically distinct Indiana serotype of VSV. The New Jersey serotype of VSV synthesized five distinct mRNA species in vitro, three of which were smaller than the corresponding species synthesized by the Indiana serotype of VSV. These included the mRNA's coding for the G, M, and NS proteins. By hybridization experiments, virtually no sequence homology was detected between the mRNA's of the two serotypes. Despite this lack of overall homology, the 12 to 18S mRNA species of both serotype contained a common 5'-terminal hexanucleotide sequence, G(5')ppp(5')A-A-C-A-G. The signicance of this finding in light of specific interactions between the two serotypes of VSV in vivo is discussed.


In vitro RNA transcription by the New Jersey serotype of vesicular stomatitis virus. II. Characterization of the leader RNA. The New Jersey serotype of vesicular stomatitis virus (VSV) was able to synthesize a small RNA (leader RNA) approximately 70 bases in length similar to the leader RNA synthesized in vitro by the genetically distinct Indiana serotype of VSV. Also, the New Jersey leader RNA contained the same 5'-terminal sequence, ppA-C-G, as the Indiana leader RNA and had a very similar base composition, with 42% AMP, 16% CMP, 18.6% GMP, and 23.4% UMP. The 3'-terminal sequence of the VSV New Jersey genome RNA was detemined and found to contain the sequence- Py-G-UOH, again the same as that of the Indiana serotype of VSV. Evidence that the New Jersey leader RNA is transcribed from the 3' end of the genome RNA was obtained from the fact that it can protect the 3'-terminal base of [3H]borohydride-labeled New Jersey genome RNA from RNase digestion. Although the New Jersey and Indiana leader RNAs were similar in many respects, they were unable to form RNase-resistant hybrids when annealed to heterologous genome RNA.


Enhanced infectivity of adenovirus type 2 DNA and a DNA-protein complex. The infectivity of adenovirus type 2 DNA and a DNA-protein complex was studied in 293 cells, a human embryonic kidney cell line transformed by sheared adenovirus type 5 DNA, and in human KB cells. Adenovirus type 2 DNA was more infectious (up to about 40-fold) in 293 cells than in KB cells, whereas a DNA-protein complex (prepared by a rapid procedure) had about the same infectivity in both cell lines. These data may mean that a factor present in 293 cells (perhaps a viral-coded protein) enhances the infectivity of free viral DNA. The infectivity of DNA and the DNA-protein complex was increased up to fivefold by brief treatment of cell monolayers with 25% dimethyl sulfoxide after transfection. Under these conditions, (i) the infectivity of native adenovirus type 2 DNA ranged from 400 to 1,300 PFU/microgram of DNA in 293 cells and from about 9 to 14 PFU/microgram of DNA in KB cells, and (ii) the infectivity of the DNA-protein complex was 6 X 10(3)to 2 X 10(4) PFU/microgram in 293 cells and 1.4 X 10(4) to 1.6 X 10(4) PFU/microgram in KB cells.


Biochemical transformation of mouse cells by herpes simplex virus type 2: enhancement by means of low-level photodynamic treatment. The biochemical transformation of thymidine kinase-deficient cells by UV-inactivated herpes simplex virus is enhanced by low-level photodynamic treatment of the infected cells. At the concentration of proflavine used, the virus was not inactivated and both virus and cellular DNA syntheses were only marginally inhibited. The observed enhancement of the transfer of a virus gene to the cell genome suggests a possible cocarcinogenic role for photodynamically active dyes at very low concentrations.


Epstein-Barr virus-associated thymidine kinase. Superinfection of Raji cells with Epstein-Barr virus induced a new thymidine kinase that was distinguishable from both adult and fetal kinases of the host cell by discontinuous electrophoresis on polyacrylamide gels and glycerol gradients.


Biochemical characterization of the amphotropic group of murine leukemia viruses. The recently described amphotropic group of murine leukemia viruses constitutes a distinct biological group, differing from the ecotropic and xenotropic groups in host range, cross interference, and serological reactivity. Viruses of this group have been detected only in wild mice from certain areas in California. By using a [3H]DNA probe synthesized in an endogenous reaction from detergent-lysed amphotropic virus (strain 1504-A), it was demonstrated that the amphotropic murine leukemia viruses are distinct biochemically, in that 20% of the viral genome sequences are not shared by AKR-type ecotropic or nay of three types of xenotropic murine leukemia virus tested. A subset of these amphotropic unique sequences, comprising one half of them, is present in the genome of wild mouse ecotropic viruses and in Moloney and Rauscher viruses as well. Sequences homologous to the entire genome of 1504-A amphotropic virus are present in the cellular DNA of all eight inbred mouse strains tested, as well as in wild Mus in Asia, in amounts varying from three to six complete viral genomes per haploid cell genome. Evidence is presented that at least 20% of the DNA sequences in both mouse- and mink-grown murine leukemia virus probes are of host-cell origin.


Characterization and classification of virus particles associated with hepatitis A. I. Size, density, and sedimentation. Virus-like particles were purified from stools of patients in an epidemic of hepatitis A in Germany. When reference MS-1 chimpanzee pre-inoculation and convalescent sera were used, the close serological relationship of the purified particles to well-known isolates of hepatitis A could be established. On the other hand, the physicochemical characteristics of the particles were determined in parallel to the characteristics of a marker parvovirus (LuIII) and a marker picornavirus (poliovirus type 2). It could be shown that the majority of the hepatitis A-associated particles band at 1.34 g/ml in CsCl and, like poliovirus, sediment at about 160S. In addition, a distinct hepatitis A antigen was observed, which banded at 1.305 g/ml and sedimented between 50 and 90S. A further component accumulated in the density range of between 1.38 and 1.44 g/ml. However, it seemed to be rather labile. Upon reisolation from CsCl and sedimentation in sucrose, it resolved into a 160S, a 90 to 100S, and a 50S form. The size of the 160S particles (27 to 29 nm) could be readily distinguished from that of the parvovirus (22 to 24 nm). It is concluded, therefore, that hepatitis A-associated virus particles are more likely to be classified with the picornaviruses than with the parvoviruses.


Characterization and classification of virus particles associated with hepatitis A. II. Type and configuration of nucleic acid. Virus particles banding at 1.34 g/ml in CsCl and sedimenting at 160S in sucrose gradients were isolated from fecal specimens of patients suffering from hepatitis. In the presence of 4 M urea and about 90% formamide, these particles released linear nucleic acid molecules of the kinked appearance characteristic of single-stranded RNA or single-stranded DNA. They could be distinguished from the nucleic acid of phage lambda added to the preparation as a marker for double-stranded configuration. Experiments in which the virus particles under investigation were incubated at pH 12.9 at 50 degrees C for 30 min revealed that their nucleic acid molecules were hydrolyzed as readily as the RNA genome of poliovirus type 2 analyzed in parallel. Both the single-stranded DNA of phage phiX174 and that of parvovirus LuIII, however, proved unaffected by this treatment, and the double-stranded DNA of phage lambda was denatured to single-stranded molecules. It was concluded, therefore, that the virus of human hepatitis A contains a linear genome of single-stranded RNA and has to be classified with the picornaviruses.


Production of unintegrated mouse mammary tumor virus DNA in infected rat hepatoma cells is a secondary action of dexamethasone. Dexamethasone, a synthetic glucocorticoid, selectively increased the rate of synthesis of mouse mammary tumor virus (MTV) RNA in clonal isolates of chronically infected rat hepatoma tissue culture cells. This hormonal effect occurred extremely rapidly and appeared to be mediated directly by the glucocorticoid-specific receptor protein. In addition to the viral RNA, unintegrated MTV DNA was also detected in these cells. Several lines of evidence are consistent with the idea that the unintegrated viral DNA is synthesized by reverse transcription of MTV RNA. (i) Unintegrated viral DNA accumulated only in the presence of dexamethasone and was produced with a time course that closely paralleled the increased accumulation of viral RNA. (ii) Density labeling of the viral DNA revealed that both strands were newly synthesized, implying a non-semiconservative mode of replication. (iii) Inhibitors of viral RNA synthesis prevented the appearance of unintegrated viral DNA. These data suggest that the production of unintegrated MTV DNA after dexamethasone treatment occurs as a secondary consequence of the hormonal induction of synthesis of viral RNA. In contrast to infected rat hepatoma cells, no unintegrated MTV DNA was detected in mouse mammary tumor cells or mouse lymphoma cells, despite the presence of high levels of viral RNA.


Attempted experimental transmission of pseudorabies virus to European starlings (Sturnus vulgaris). Forty European starlings (Sturnus vulgaris) were inoculated orally with pseudorabies virus. Subsequent attempts to isolate the virus from feces, liver, lung and kidney of the birds were negative; antibody against the virus could not be detected in their serums.


Viral inclusions in raccoon liver cells. Three young raccoons (Procyon lotor), two from Michigan and one from Arizona, died suddenly from acute infections. Intranuclear inclusion bodies and viral particles typical of herpesvirus were seen in liver cells from all three. Inclusions also were seen in the nuclei of endothelial cells in the lung, liver, glomeruli and reticuloendothelial cells of the spleen. The source of the infection was not determined, but possible transmission from other species could not be ruled out.


Oral polio vaccine. Effect of booster vaccination one to 14 years after primary series. We studied the persistence of antibody after vaccination and the response to booster revaccination with trivalent oral polio vaccine (TOPV) administered at varied intervals after the primary series in a large group of children. Decline in antibody was related to intervals since last vaccination, and not to sex, age, age at primary vaccination, or type and number of previous administrations. Geometric mean titers of neutralizing antibody were 11.3 for type 1 and 8.0 for types 2 and 3 poliovirus when vaccine had been given within the previous year, declining to 3.2, 3.0, and 2.1 for types 1, 2, and 3 after nine years. Most children with an initial titer of 4 or less responded to revaccination with a fourfold or greater increase in titer of IgG. Geometric mean titers for all three types of polio dropped to this level when last TOPV administration had been five to six years or more.


Scintiangiography of hepatic masses in childhood. Forty-three children with intrahepatic masses had technetium Tc 99m sulfur colloid hepatic scintigrams including scintiangiography. Hypervascularity occurred in nearly one third of the masses, but was limited to children with primary liver tumors including hepatomas, hepatoblastomas, hepatic adenomas, and cavernous hemangiomas. All metastases, abscesses, and hematomas were hypovascular. Routine inclusion of scintiangiography may allow separation of primary hypervascular liver tumors from other hypovascular masses whose static hepatic scintigrams are virtually identical.


Carcinoma of the lung and cigarette smoking. Effect on serum ribonuclease activity. Serum ribonuclease levels were determined in 54 patients with lung carcinoma, 74 long-term cigarette smokers, and 172 nonsmokers. The mean serum ribonuclease level was significantly higher in patients with lung carcinoma and long-term smokers compared with healthy nonsmokers (P less than .001). The serum ribonuclease activity level was not related to chronological age, sex, or race of the smoker or nonsmoker population. Forty (75%) of 53 patients with lung cancer and 49 (66%) of 74 smokers had elevated serum ribonuclease levels compared with 13 (7%) of 179 nonsmoker healthy controls (P less than .001). The highest incidence of elevation was noted in patients with epidermoid carcinoma (95%).


Studies on a dyslipoproteinemia in hemodialysis patients. Grossly abnormal serum lipoprotein (Lp) profile which appeared as a broad-midband pattern (BMP) in PAG electrophoresis was detected in 67% of uremic patients maintained by prolonged hemodialysis therapy (hemodialysis patients). It was revealed that BMP was ascribed to the presence of a discrete series of Lps which were intermediate between beta-migrating LDL and pre-beta-migrating VLDL regarding buoyant density, Sf rate, chemical composition, and behavior in electrophoresis. The results in the present study indicate that the accumulation of these Lps, which may include the previously-known Lp species such as the intermediate-density Lp or the remnant particles, is taking place in hemodialysis patients.


Effects of intraventricularly administered polyamines spermidine and spermine on sleep-wakefulness cycles in rats. Freely moving rats with chronically implanted electrodes were used and the electroencephalogram (EEG) from the occipital cortex and the dorsal hippocampus, the electromyogram (EMG) and the electrooculogram (EOG) were simultaneously recorded. The REM (rapid eye movement) sleep stage was significantly shortened without a marked change of the Non-REM sleep from the 24th to the 32nd hr following injection of 40 microgram of SPD. The waking stage significantly increased, and the Non-REM sleep and REM sleep markedly decreased immediately after the injection of 40 microgram of SPM to the 8th, and from the 24th to the 32nd hr. The amount of total sleep significantly shortened from the 72nd to the 80th hr following injection of 40 microgram of SPD. The administration of 40 microgram of SPM significantly reduced the amount of total sleep in every period measured  for 8 hr. Marked change was observed from the 24th to the 32nd hr following injection of 100 microgram of PUT, and Non-REM sleep and total sleep produced a significant decrease. These results show that the intraventricular administration of polyamines including PUT inhibits the amount of normal sleep.


Effect of enkephalin and substance P on sympathetic nerve transmission in mouse vas deferens. Effect of methionine-, leucine-enkephalin (met-, leu-enkephalin) and substance P on the transmission in mouse vas deferens was studied. Both met- and leu-enkephalin inhibited electrically induced contraction of vas deferens at 10(-8)-10(7) M, met-enkephalin being 1.4 times more active than leu-enkephalin. Nalorphine (10(-6) M) antagonized these effects. Substance P (10(-9)-10(-7) M) had no effect on the contraction. Met- and leu-enkephalin (10(-7)-10(-5) M) decreased the high potassium induced [3H]-norepinephrine release from vas deferens, while substance P (10(-6) M) significantly increased it. Nalorphine (10(-5) M) reversed the inhibitory effect of met-enkephalin. These results indicate that these peptides modify the transmission of sympathetic nerve in mouse vas deferens.


No evidence for involvement of dopaminergic receptors in the positive inotropic action of dopamine on the isolated rabbit papillary muscle. Experiments were carried out on the isolated rabbit papillary muscle driven at 0.5 Hz in order to further elucidate the mechanism of the positive inotropic effect evoked by dopamine. The dose-response curve for dopamine was not affected by the antagonists pimozide (10(-6) M), yohimbine (10(-5) M) pindolol (3 x 10(-8) M) and phentolamine (10(-6) M) when these agents were given separately. Only the simultaneous administration of yohimbine plus pindolol and phentolamine plus pindolol, respectively, shifted the entire curve to the right. This shift was not further influenced by pimozide. Dopamine (10(-4) M) increased the cyclic AMP content of the papillary muscle by about 50%; this increase was not affected by pimozide, but was markedly elevated by yohimbine and completely depressed by pindolol. From the present results it is concluded, that dopamine produces its positive inotropic effect through stimulation of myocardial alpha-as well as beta-adrenoceptors to about the same degree; stimulation of specific dopaminergic receptors, however, is not involved. The stimulation of beta-adrenoceptors is accompanied by an increase of the cyclic AMP level, while that of alpha-adrenoceptors is not.


Morphine-induced changes in cyclic AMP metabolism and protein kinase activity in the brain. The effects of consecutive oral administration of morphine on the cyclic AMP synthesizing system and cyclic AMP dependent protein kinase activity in the cerebral cortex of mice were examined. The administration of morphine (2--4 weeks) induced an increase of the cyclic AMP formation by activating adenylate cyclase, whereas responses of the cyclic AMP synthesizing system to biogenic amines (norepinephrine, dopamine and histamine) added in vitro was found to be significantly attenuated in these animals. Cyclic AMP dependent protein kinase activity in the cerebral cortex was also increased following a consecutive oral administration of morphine. These changes in the activities of adenylate cyclase and protein kinase were found mainly in crude mitochondrial and/or synaptosomal fractions. Morphine induced decrease in the response of the cyclic AMP synthesizing system to biogenic amines was rapidly reversed, and a significant increase of the cyclic AMP formation in the presence of added norepinephrine compared with that found in morphinized animals was observed following the administration of levallorphan, a narcotic antagonist. On the other hand, the changes in adenylate cyclase and cyclic AMP dependent protein kinase activities were not affected significantly by levallorphan administration. These results suggest that alterations in activities of cyclic AMP synthesizing system and of cyclic AMP dependent protein kinase may be involved in processes of the formation of morphine dependence. Possible involvement of abrupt increments in the sensitivity of "norepinephrine receptor-adenylate cyclase" system and a subsequent increase in cerebral cyclic AMP is also suggested as a cause of morphine withdrawal syndrome.


[Metastasis from the colon to the choroid (author's transl)]. Various investigators have concluded that metastatic carcinoma is the most common malignant tumor of the eye. In about 70% of all cases the primary site is breast or lung. Metastasis from the colon to the choroid is found very seldom. The clinical course in our case is described and the elevated fashion of the right eye's metastatic carcinoma is shown. Usually cancerous tissue in the choroid spreads and produces a rather flat, diffuse lesion. Problems in making the diagnose are discussed. In choroidal metastasis radiation should be attempted, for adequate vision can sometimes be maintained for the remaining time of the patient's life.


[Considerations on the meta-herpetic keratitis (author's transl)]. The meta-herpetic keratitis is clinically characterized by the occurrence of a parenchymatous keratitis due to iterative herpetic corneal wounds. The rupture of the Bowman membrane which makes such a deep wound possible is performed by an enzyme: collagenase. This parenchymatous keratitis is rarely due to an extension of the viral infection. Most often it has an immunologic origin. It is a disciform keratitis due to a viral allergy, or a polymorphic keratitis connected with a bacterial, often tubercular, origin. In this case, one always notices a characteristic sugar tongs like composite limbic vascularization. Recovery can only be obtained by a specific desensitization.


[Coronary heart disease in patients with primary type V hyperlipoproteinemia (author's transl)]. In two patients with primary Type V hyperlipoproteinemia with typical clinical features including recurrent bouts of abdominal pain a myocardial infarction was diagnosed. In both cases coronary angiography revealed a severe three vessel disease. The case reports demonstrate that the incidence of ischemic heart disease in patients with Type V hyperlipoproteinemia is higher than reported in the literature. In each case of severe abdominal pain, even in younger Type V patients, a myocardial infarction has to be excluded, In both patients a selective depression in the activity of lipoprotein lipase was found. The possible pathogenetic implication of this finding will be discussed.


[On the ACTH-corticoid relation in essential hypertension in dependence of plasma-renin activity (PRA) (author's transl)]. Patients with essential hypertension were studied for the reaction of the hypophyseal-adrenal-system before and during insulin hypoglycemia test. We found in our results that the mean total corticoid levels in plasma of hypertensive patients with low or high PRA are significantly higher than control levels. In insulin hypoglycemia test the mean corticoid levels of patients with normal and high PRA do not differ from the mean levels found in normal individuals whereas hypertensive patients with low PRA have significant lower plasma corticoid levels. The mean baseline and hypoglycemia induced plasma ACTH levels of each group of hypertensive patients are higher than those of the controls. Patients with high PRA show the highest rise of mean plasma ACTH levels during hypoglycemia. These data suggest that the adrenal system of hypertensive patients produces less total corticoids; plasma ACTH levels of these patients therefore are higher than those of normal individuals.


Outpatient aftercare as a factor in treatment outcome; a pilot study. Outpatients receiving psychotherapy at the facility in which they had previously been inpatients were more likely to remain abstinent than were former inpatients receiving outpatient care elsewhere.


Aftercare for alcoholics: services of community mental health centers. Fewer than half of the patients hospitalized for alcoholism treatment received aftercare from a community mental health center, the variables significantly related to receipt of aftercare being number of previous hospitalizations and referral to the hospital by self or family rather than by a court.


[Surgery of extensive glomus jugulare tumors (author's transl)]. Clinical and neuroradiological aspects of extensive glomus jugulare tumors are presented. Surgery of these tumors is based on their growth along the veins: caudally along the internal jugular vein, cranially along the sigmoid sinus and medially along the transbasilar veins up to the cavernosus sinus. The main problem in total removal of glomus jugulare tumors is the relieving of the internal carotid artery from the tumor, especially in its intrapyramidal part. It is well known that glomus jugulare tumors tend to bleed excessively during surgery. Therefore the branches of the external carotid artery and the transverse sinus are ligated at the beginning of the operation. To avoid infection from the Eustachian tube to the posterior fossa, to close the dural defect over the cerebellar hemisphere and to protect the internal carotid artery a pedicled flap of the sternocleidomastoid muscle, based cranially, is prepared and rotated upwards in the base of the skull. 9 cases of extensive glomus jugulare tumors are presented.


Cushing's syndrome due to adrenal adenoma with persistent diurnal cortisol secretory rhythm. A 41-yr-old female with presumed Cushing's syndrome was found to have a diurnal cortisol rhythm characterized by low values of 8:00 a.m. and consistently high values at 4:00 p.m. and midnight. Hourly sampling of plasma cortisol over 24 hr confirmed this rhythm, as did measurement of urinary free cortisols in samples collected every 6 hr over 24 hr. Hypercortisolemia was not suppressed by 2 mg of dexamethasone given every 6 hr for 24 hr. The adrenal tissue was responsive to ACTH. Iodocholesterol scanning revealed unilateral activity, and the patient's syndrome was cured by resection of an adrenal adenoma. In this patient a diurnal cortisol secretory pattern was present due to the secretory activity of the adenoma. The cause of the abnormal but persistent diurnal pattern is unknown.


Thiazide diuretics do not potentiate cAMP response to parathyroid hormone. We evaluated the hypothesis that thiazide-induced hypercalcemia reflects potentiation of the cAMP response to parathyroid hormone (PTH) consequent to inhibition of phosphodiesterase in bone and kidney. A panel of thiazide diuretics did inhibit low-Km phosphodiesterase activity from bone homogenates. However, furosemide, a nonthiazide diuretic that does not promote calcium retention, was more potent a phosphodiesterase inhibitor than either chloro- or hydrochlorothiazide (CTZ, HCTZ). Thiazides did not influence basal or PTH-stimulated cAMP levels in incubated calvaria or renal cortical slices. Administration of CTZ or HCTZ to rats for 4 days did not affect basal cAMP, nor did such treatment potentiate the cAMP response in Calvaria to infusion of parathyroid extract in vivo. CTZ, HCTZ, and furosemide increased basal adenylate cyclase from renal cortex but did not affect PTH-stimulated activity. Adenylate cyclase from bone was not affected by thiazides but was inhibited by furosemide. Thiazide treatment potentiated the calcemic response to parathyroid extract in vivo but did not affect the calcemic response to dibutyryl cAMP. We conclude that potentiation of the cAMP response to PTH does not underlie the unique effects of thiazides on calcium metabolism.


[Effect of histones on a change in the enzymatic activity of Escherichia coli and Staphylococcus aureus cells]. Histones have been found to interfere with the permeability of the cellular membranes of Escherichia coli and Staphylococcus aureus, this resulting in the liberation of 14C-amino acids, the UV-absorbing endogenous material, and the products of RNA degradation. Histones, particularly arginine-rich fractions, stimulated the RNAase activity of the bacteria. Histones inhibited the phosphatase activity of E. coli and stimulated that of Staphylococcus aureus. The ATPase activity of the cells increased upon contact with histones. These enzyme systems are presumed to be components of a mechanism whose activity increases upon the action on the cells of factors interfering with the permeability of the membranes so that low-molecular-weight compounds are liberated from the cells. It is possible that the RNAase, ATPase and phosphatase systems repair the pool of low-molecular-weight compounds, thus contributing to the survival of the bacteria.


[Changes in the electron transport chain in Escherichia coli depending on the cultivation conditions and growth phase]. Changes in the electron transport chain of E. coli K-12 were studied as a function of the growth phase and the nature of a terminal electron acceptor in the growth medium. The content of flavins in the preparations of bacterial membranes hardly changed in all cases. The highest concentration of quinones was observed in the bacterial membranes at the stationary growth phase under anaerobic conditions of growth in the presence of nitrate. These membranes contained also the greatest amount of cytochrome beta1. The concentration of cytochrome alpha2 in all the membranes was low and varied among different preparations. All the membranes contained a CO-binding pigment whose content was maximal in the membranes of "nitrate" cells. The membranes of cells grown under aerobic conditions oxidized malate, apart from NADH and lactate, whereas the membranes of cells cultivated under anaerobic conditions in the presence of nitrate oxidized formiate. In most cases, the oxidase activity of the membranes of cells collected at the stationary growth phase was higher cf. the exponential phase.


[Fatty acid makeup of Escherichia coli cells with repressed and derepressed phosphohydrolase biosynthesis]. Fatty acid composition of the cells of Escherchia coli wild strains K-10, and K-12 and the mutants of the regulatory genes for alkaline phosphatase was studied in conditions of repression and derepression of biosynthesis of phosphohydrolases. Derepression of phosphohydrolases was not accompanied with specific changes in fatty acid composition of the cells. An increase in the content of cyclopropanic acid in conditions of phosphorus deficiency and a decrease in the level of unsaturated fatty acids are related to deceleration of growth of the cells in these conditions.


[Comparison of liposoluble and water-soluble contrast media in sialography]. Highly, averagely and lightly viscous contrast media most frequently used in radiography of the salivary glands have been compared. It is concluded that the preparation which has so far given the best results is liposoluble but averagely viscous and not too contrasting.


A kinetic analysis of glucokinase and glucose-6-P phosphatase in Dictyostelium. Using a mathematical model of carbohydrate metabolism in Dictyostelium discoideum, the kinetic expressions describing the activities of glucokinase and glucose-6-P phosphatase have been analyzed. The constraints on the kinetic mechanisms and relative activities of these two enzymes were investigated by comparing computer simulations to experimental data. The results indicated that, (1) glucose-6-P is compartmentalized with respect to the enzymes involved in glucose-60P, trehalose and glycogen metabolism, (2) a differences of approximately 0.6 mM/min in maximum specific activity of glucokinase compared to glucose-6-P phosphatase is required in order for the model to produce end product carbohydrate levels consistent with those observed experimentally, (3) the Km of glucokinase for glucose strongly influences the steady state levels of glucose in the absence of external glucose, and (4) changing the order of product removal in the reaction catalyzed by glucose-6-P phosphatase influences the level of glycogen and trehalose.


The use of viable hepatocytes to study the hormonal control of glycogenolysis in the chicken. The rapid isolation of high yields of parenchymal cells from chicken liver is described. Stringent tests of viability show that the isolated hepatocytes are both structurally and metabolically similar to those in intact liver. During incubation viability decreased and the significance of this change on the interpretation of metabolic experiments is discussed. Lactate was a much more effective gluconeogenic precursor than pyruvate even in the presence of additional reducing equivalents. Hepatocytes isolated from fed chickens produced glucose from glycogen degradation. Glycogenolysis was stimulated by glucagon, dibutyryl cyclic AMP and adrenaline. Half maximal glucagon effects were elicited by physiological concentrations of the hormone. Glucagon and dibutyryl cyclic AMP stimulated glucagon, dibutyryl cyclic AMP and adrenaline their action was not additive to that of adrenaline.


Periodicity and fragment size of DNA from mouse TLT hepatoma chromatin and chromatin fractions using endogenous and exogenous nucleases. The action of micrococcal nuclease, DNase I and DNase II on mouse TLT hepatoma chromatin revealing the periodicity of its structure as visualized by denaturing and non-denaturing gel electrophoresis, was consistent with the action of these enzymes on other chromatins. Micrococcal nuclease showed a complex subnucleosome fragment pattern based on multiples of 10 base pairs with a prominant couplet at 140/160 base pairs and the absence of the 80 base pair fragment. This couplet of the core and minimal nucleosome fragments was conspicuously present in the mononucleosomes found in the 11S fractions of a glycerol gradient centrifugation. DNase I and II produced a fairly even distribution of a 10 base pair increasing series of fragments to about 180 base pairs, a pattern also repeated in the DNA of nucleosome glycerol-gradient fractions. In limited digestions by these nucleases multinucleosomic DNA fragments are pronounced. These fragment lengths are multiples of an estimated average repeat length of nucleosome DNA of 180 base pairs. The action of the endogenous Mg/Ca-stimulated endonuclease produced only limited cuts in the hepatoma chromatin resulting primarily in multi-nucleosomic DNA fragment lengths and only upon lengthy digestion limited subnucleosomic, 10-base-pair multiple fragments are produced. The putative euchromatin-enriched fractions (50-75S) of the glycerol gradient centrifugation of autodigested chromatin, similarly, contained primarily the multinucleosomic DNA fragment lengths. These results are consistent with our previous electron microscopic demonstration that autodigested chromatin as well as the putative euchromatin-enriched fractions were composed of multi-nucleosomic chromatin segments containing a full complement of histones.


[Circadian rhythm of cortisol under ACTH and dexamethasone therapy of convulsions in early childhood (BNS- and Lennox-syndrome) (author's transl)]. Convulsions in early childhood were treated successfully in a long term and high dosage regime with depot corticotrophin (80-120 IU/die) and dexamethasone (0.3 mg/kg/die). Depot-beta1-23corticotrophin stimulated, dexamethasone suppressed plasma cortisol levels, which, accordingly, ceased to show circadian rhythms. If dexamethasone was given alternately (every second day) and in a reduced dose (0.1 mg/kg) plasma cortisol levels increased, and if dosage was reduced again to 0.05 mg/kg cortisol levels showed circadian rhythms comparable to those before therapy. There was no disturbance of the hypothalamic--pituitary--adrenal system at the end of therapy. Considering experimental results in animals corticotrophin peptides and dexamethasone molecules might have a direct effect on the brain as far as cerebral convulsions are concerned. Because of serious side effects of steroids during treatment it seems to be worth while to try corticotrophin fragments, which had similar effects on the brain as beta1-24corticotrophin in animals, but without stimulating the adrenals.


[Physico-chemical properties of the RNA of Sendai virus. III. Interaction with proflavin]. The binding isotherms of the Sendai virus single- and double-stranded RNA-proflavine complexes have been studied. The existence of two regions on the binding curves, corresponding to two subtypes of the strong complex (I1 and I2) has been demonstrated. The association constants and the numbers of binding sites for both subtypes were determined as a function of ionic strength. Both of the single- and for the double-stranded RNA the association constant for I1 complex were higher than those for I2 complex under all ionic strengths conditions. For the double-stranded RNA the variation of the ionic strength is more importance in case of the I1 complex formation. The total number of binding sites increases with a decrease of ionic strength. At low ionic strength (10(-4) M NaCl) the number of binding sites for single- and double-stranded RNA is practically the same and is equal to the number of binding sites for DNA at high ionic strength (1 molecule of proflavine per 3 nucleotides pairs). The heat denaturation of the RNA-proflavine complexes under different ionic conditions has been also investigated. The melting curves for double-stranded RNA-proflavine complex had two waves at high and low ionic strengths. For the single-stranded RNA the high temperature wave occured only at the high ionic strength. The dependence of heat denaturation of single-stranded RNA on ionic strength was examined for the evaluation of RNA structure. In these experiments a significant decrease of the width of melting interval under low ionic strength conditions was observed. It may reflect the existance in the RNA molecule of long helical regions. The occurence of such structures is likely to be responsible for an increase in the number of binding sites at low ionic strength and also for the appearance of the second wave on the melting curves of the single-stranded RNA-proflavine complexes at high ionic strength.


[Comparison between combination therapy with clofibrate and beta-pyridylcarbinol and clofibrate monotherapy (author's transl]. In order to determine whether the lipid-lowering effect of combined treatment with clofibrate and beta-Pyridylcarbinol exceeds that of clofibrate monotherapy, a double-blind crossover study was performed. 17 patients with primary hyperlipoproteinemia of Type IIa and 10 patients with primary hyperlipoproteinemia of Type IIb received either Lipofacton (1.000 mg clofibrate and 50 mg beta-pyridylcarbinol per day) or clofibrate (1.500 mg per day) for a period of 6 weeks each. Before beginning therapy and between both periods of medication, placebo was administered for 14 days. In both total plasma and in the LDL fraction the cholesterol level was lowered by less than 10% for both substances. In patients with hyperlipoproteinemia of Type IIb, the triglyceride levels were lowered by about 40%.


[Distribution of cytomegalovirus in children and adults in the Munich area (author's transl)]. In a regional contamination study 798 healthy persons from the Munich area were examined for cytomegalovirus antibodies. The highest proportion of antibodies (60%) was found in neonates in the first days of life. At the age of 6 to 24 months, only 10% of the children were still seropositive. At this time the maternal antibodies were no longer important. In late adult life, 57% of those examined possessed antibodies against the virus. An attempt was also made to isolate the cytomegalovirus from the urine of 157 healthy persons. This isolation was only possible in three apparently healthy infants aged from 3 to 12 months.


Unscheduled DNA synthesis induced in mouse spermatids after combined treatment with methyl methanesulfonate and X-rays. Unscheduled DNA synthesis (UDS), which is considered to be DNA repair, has been studied in early- to mid-spermatid stages of the mouse after combined treatments with X-rays and methyl methanesulfonate (MMS). UDS in spermatids was detected by giving testicular injections of [methyl-3H]thymidine ([3H]dThd) and making use of the fact that no scheduled DNA synthesis occurs in the germ cells after the last S period in primary spermatocytes. X-rays and MMS are each able to induce UDS in mouse spermatids. However, there was a statistically significant reduction in the amount of UDS observed when X-ray exposures of from 200 to 600 R were given 4 h before an i.p. injection of 75 mg/kg of MMS and concurrent testicular injections of [3H]dThd. This reduction in UDS is more than can be explained by the completion of repair of X-ray-induced DNA lesions. We suggest that the reduction in UDS is the result of an X-ray-produced impairment of a least a part of the repair mechanism involved in correcting MMS-induced DNA lesions. When the time interval between a 600-R X-ray exposure and MMS treatment was between 3 and 20 h (latest time interval s;udied) there was a statistically significant reduction of UDS in the spermatids. No significant decrease in UDS response occurred when the time interval between radiation exposure and MMS treatment was less than approximately 3 h.


Hyperbaric oxygen and radiation therapy in the management of glioblastoma. A pilot clinical trial on radiotherapy of glioblastoma with and without hyperbaric oxygen was performed at the Columbia-Presbyterian Medical Center. Eighty previously untreated patients with histologically proved glioblastoma were evaluated; 38 were irradiated under hyperbaric oxygen and 42 (controls) in atmospheric air. The survival rates were calculated according to the actuarial analysis method. At the end of 18 months, the survival rate appeared considerably higher in the oxygen group (28%) than in the controls (10%). At the end of 36 months, no patients in the control group survived, whereas 2 patients in the oxygen group were alive beyond 45 and 48 months, respectively. The median survival time was 38 weeks for those treated under oxygen and 31 weeks for the air control group. Owing to the small population samples and the pilot nature of this study, the difference in survival rates between the two groups was not statistically significant. The toxicity of hyperbaric oxygen was well tolerated by most patients, and the quality of survival in the hyperbaric oxygen group was equal to or slightly better than that of the control group. This pilot clinical study paved the way for further controlled clinical trials of hyperbaric oxygen and oxygen-mimicking drugs, including the electron-affinic compounds that could have differentially sensitized the hypoxic tumor cells.


Treatment of an intracerebral rat brain tumor with Corynebacterium parvum and radiation. Rats bearing intracranial brain tumors were treated with single ip injections of the killed Corynebacterium parvum, a single dose of X-rays, or a combination of both treatments. Animals given injections of 2.6 mg C. parvum 12 days after implantation of the tumor had a median survival time (MST) of 50 days compared with an MST of 44 days for an untreated group. These animals given C. parvum had a 33% increased life-span (ILS) that was significant at the P=0.05 level. Although the MST of animals that received C. parvum 2 days before a tumor X-ray dose of 3,015 rads on day 14 was greater than that of rats given irradiation alone, the ILS of the group administered both treatments was not significant relative to the latter group. The MST of animals that received C. parvum 7 days before irradiation was not different from that of animals given injections of C. parvum 1 day postirradiation.


Chemical- and virus-induced brain tumors. Experimental animal models resembling most human brain tumor types can be induced by exposure to oncogenic viruses or chemical carcinogens: Astrocytomas and glioblastoma multiforme can be produced experimentally by intracerebral injection of oncornaviruses, whereas medulloblastomas, choroid plexus papillomas, and ependymomas can be induced by the papovaviruses. Adenoviruses have been utilized to cause medulloepitheliomas, neuroblastomas, and retinoblastomas. All three groups of viruses can result in sarcoma production. Gliomas represent the primary tumor type induced in the brain by chemical carcinogens. These autochthonous tumor systems are reviewed, with emphasis on methods, tumor type, latency period, advantages, and disadvantages. In addition, recent investigations of molecular events involved in neoplastic transformation by chemical carcinogens are summarized.


Nomenclature for gliomas.

Nomenclature for gliomas. Gliomas, derived from astrocytes, oligodendroglia, or ependyma, are each united into a continuum by a graduation of anaplasia. Neoplasms originate at all levels of each continuum; subsequently, some move along its declivity. Conversely, neuroblastic tumors may differentiate, whereas concomitantly in the same lesion, the glial stroma may dedifferentiate. Anaplastic glia, as in a glioblastoma multiforme, can initiate malignant transformation in alien cells.


Pathologic tumor type and response to treatment. The general biology of the common primary brain tumors is reviewed, as are the results of surgery, radiation therapy, and chemotherapy for these neoplasms. The problems of classification, particularly of the astrocytomas, are noted, with the resultant difficulties in comparing different series. The development and consistent application of a widely acceptable nomenclature for brain tumors would be useful in the comparison of various therapies and for ease in communication. Further, a better general understanding of the biology of the various brain tumors is desirable. Reliable data relative to old or new forms of therapy seem to require national or regional studies in view of the relative rarity of some of these tumors. Except for medulloblastoma, the best results of therapy thus far are only slightly encouraging. We must also be aware that some of our treatments may do as much harm to some patients as their tumors; therefore, we must all strive for a high autopsy rate so that we can monitor our therapy and recognize unexpected complications as quickly as possible.


Selected observations on the epidemiology of pharyngeal cancers. The epidemiology of nasopharyngeal cancer (NPC) has been compared with that of other pharyngeal cancers in Los Angeles County. Epstein-Barr virus (EBV) antibody titers were elevated in Caucasians with pharyngeal squamous cell carcinomas regardless of subsite. As expected, an excess of NPC cases was found among Chinese. The role of various etiologic factors in NPC including EBV, inhaled carcinogens, and salted fish was discussed.


Indirect complement fixation test with foot-and-mouth disease virus antigen concentrated by polyethylene glycol precipitation. To a BHK-21 cell culture fluid infected with the O, A, or Asia 1 type of foot-and-mouth disease (FMD) virus was added polyethylene glycol 16000 to a concentration of 10% (w/v). Then the fluid was concentrated to one-tenth of the original volume. The resulting concentrated virus antigens showed a complement fixation (CF) titer ranging from 12 to 14. The rate of recovery of CF activity was in a range of 40 approximately 80%. Each antigen was applied to the indirect complement fixation (ICF) test with serum from cattle infected experimentally with the respective type of virus to estimate antibody titers. When antibody was examined for a rise and fall, it began to be detected 4 approximately 7 days after inoculation and was detectable even 63 days after inoculation. It showed a tendency to exhibit a rise and fall parallel with that of neutralizing antibody, although it was always lower in titer than this antibody. ICF antibody corresponding to one type of virus was type-specific, presenting little crossing with a heterologous antigen. As a result, it was clarified that such antigen as prepared from an infected cell culture fluid after concentration with polyethylene glycol was applicable to the ICF test for the estimation of the titer of antibody against FMD virus.


Interactions of opiates and prostaglandins E with regard to cyclic AMP in striatal tissue of rats in vitro. The effects of prostaglandins E on the concentration of cyclic AMP (cAMP) and a possible antagonism of opiates vs. prostaglandins E were studied in homogenates and in slices of rat striata in vitro. In homogenates, PGE1 or PGE2 did not affect the synthesis of cAMP. Morphine slightly lowered the cAMP synthesis, in presence or absence of PGE1 or PGE2. In slices, PGE2 significantly elevated the cAMP concentrations, either in presence or in absence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine. Morphine, met-enkephalin and levorphanol, but not dextrorphan, antagonized this rise of cAMP. The effect of morphine was antagonized by naloxone. Adenosine or an elevation of K+-ions raised the cAMP concentrations, and PGE2 induced a further increase. In presence of elevated K+-ions or adenosine, however, morphine did not antagonize the PGE2-induced rise of cAMP concentration. It is suggested that under some experimental conditions described in the literature, endogenous activators of cAMP formation, e.g. adenosine, might mask the inhibitory effect of opiates on stimulation of opiates on stimulation of cAMP synthesis induced by prostaglandins E.


ACTH-induced lipolysis in rat adipocytes: structure-activity relationships. The lipolytic action of natural porcine ACTH1(-39) and of a number of highly purified synthetic ACTH peptide fragments was studied using rat adipocytes. Of the analogues tested, only ACTH1(-24) exhibited full lipolytic activity with respect to intrinsic activity and affinity. Several shorter fragments appeared to be full agonists but had lower affinity. Fragments ACTH5(-10) and ACTH7(-10) were inactive. No antagonistic effects against the lipolytic action of ACTH could be demonstrated with substimulatory doses of ACTH1(-16), ACTH1(-10), ACTH7(-24) and ACTH11(-24). Based on the relative potency derived from dose-response curves, a more refined model with respect to the active centers being encoded in various sequences of the hormone, is proposed.


In vitro regulation of ACTH release from neurointermediate lobe of rat hypophysis. I. Effect of crude hypothalamic extracts. Using incubated glands, we showed that cerebral cortex and liver extracts (CCE and LE) stimulated ACTH release from neurointermediate lobe (NIL) of hypophysis as well as hypothalmus extract (HE) did. Moreover, the HE-induced ACTH release was much smaller for the NIL (1.9 X basal level) than for the anterior lobe (AL; 13.7 x basal level). Thus, under these conditions, HE seemed to have no specific effect on NIL ACTH release. Using superfused glands, we showed: (a) that both spontaneous and HE-induced ACTH release decreased during superfusion; (b) that using this system, a specific stimulatory effect on HE on NIL was observed. In contrast to HE, CCE and LE had only a small effect on NIL ACTH release (always less than 20% of that caused by HE) which could be considered as a nonspecific response; (c) that trypsin suppressed the stimulating effect on HE as well on NIL as on AL; and (d) that arginine antidiuretic hormone (ADH) was not responsible for the stimulating effect of HE on NIL ACTH release, because synthetic ADH had no effect and HE containing ADH (from normal rats) or HE containing no ADH (from Brattleboro rats or from immunoneutralization of ADH in normal HE) had the same effect. From these results, we can conclude that HE contain a peptidic factor different from ADH which is able to stimulate in vitro release of ACTH from the NIL.


Effect of spinal cord transection on the endocrine and blood pressure responses to intravenous clonidine. To determine whether the inhibitory effect of clonidine (CLON) on renin secretion is due in part to a direct action on the kidneys or due entirely to an action on the brain, the drug was administered intravenously in a dose of 30 microgram/kg to dogs in which the spinal cord had been transected in the cervical region. Renal perfusion pressure was held constant by adjusting a suprarenal aortic clamp. The decrease in plasma renin activity produced by CLON in dogs with intact spinal cords was abolished, and in 5 of 8 dogs tested, plasma renin activity rose. The decrease in blood pressure seen in control dogs was replaced by a prolonged pressor response. The ACTH response, as measured by plasma corticoids, and the growth hormone (GH) response were not significantly reduced. The data indicate that at least at this dose, the depressor response and the decrease in renin secretion produced by CLON are completely central in origin.


An improved radioimmunoassay for alpha-melanocyte-stimulating hormone (alpha-MSH) in the rat: serum and pituitary alpha-MSH levels after drugs which modify catecholaminergic neurotransmission. A modification of our previous radioimmunoassay (RIA) for alpha-melanocyte-stimulating hormone (alpha-MSH) is described that permits the measurement of circulating levels in the rat without the need for an extraction procedure. Using this method, serum and neurointermediate lobe (NIL) immunoreactive alpha-MSH levels were measured in rats after administration of haloperidol, 2-bromo alpha-ergocryptine (CB 154), and alpha-methyl-p-tyrosine (alpha-MPT). Haloperidol caused a rapid increase, alpha-MPT a slow increase, and CB 154 a rapid decrease in serum alpha-MSH. At the time intervals studied none of the drugs had any significant effect on NIL alpha-MSH content. It was concluded from the results of the above drug treatments that modulation of dopaminergic neurotransmission both pre- and postsynaptically produces changes in serum immunoreactive alpha-MSH levels. This supports the suggestion that circulating alpha-MSH in the rat is under an inhibitory control by a catecholaminergic system.


Inhibition of corticotropin secretion by hypothalamic administration of indomethacin. The importance of endogenous prostaglandins (PGs) in regulating CRF release was investigated by administering the PG synthesis inhibitor, indomethacin (Ind), in specific hypothalamic regions to adult female rats. Solid Ind pellets placed in the anterior hypothalamic area (AHA) significantly reduced the elevated levels of plasma corticosterone normally observed after the surgical stress of the stereotaxic procedure. Similar pellets placed in the hypothalamic median eminence region (ME) completely blocked the normal compensatory hypertrophy observed 48 h after unilateral adrenalectomy. Plasma corticosterone secretion in response to hemorrhage (1% b.w.) or laparotomy with intestinal manipulation (LAP) in dexamethasone pretreated rats was reduced by Ind pellets, or Ind in phosphate buffer (BUF), respectively, implanted 2 h prior to the stress. These data suggest that the PGs may be important in mediating CRF release in response to a variety of stimuli.


Effects of hypothalamic lesions and drugs interfering with dopaminergic transmission on pituitary MSH content of rats. The physiological significance of the dopaminergic innervation of the pars intermedia was studied in rats. Electrothermic lesions were made in the mediobasal hypothalamus in order to destroy all hypothalamo-hypophysial connections. Pituitary MSH content decreased to 25% of control values 8 h after lesioning. From the 1st day, however, pituitary MSH content gradually increased, reaching control levels after about 1 week. Haloperidol and pimozide (both neuroleptics) were used to block specifically dopamine (DA)-receptors. Single administration of these drugs induced a time and dose dependent reduction in pituitary MSH. A maximal decrease to about 60% was found 4-5 h after i.p. administration of 2.5 mg/kg haloperidol and 1.0 mg/kg pimozide. Apomorphine and 2-Br-alpha-ergocryptine (DA-receptor stimulating drugs), had no effect on pituitary MSH stores of intact rats, but completely prevented the lesion-induced fall in pituitary MSH content. The results show that DA-receptors located within the pituitary itself are involved in the control of MSH release indicating that the effect of hypothalamic lesions on pituitary MSH content is primarily caused by interruption of dopaminergic neurotransmission in the hypophysis. We conclude therefore that the dopaminergic arcuato-hypophysial neurones innervating the pars intermedia tonically inhibit the release of MSH in intact rats.


Reduced neuromuscular transmission safety factor in multiple sclerosis. Regional intravascular injections of d-tubocurarine (0.3 mg) were administered in the hands of 23 normal subjects and 19 patients with multiple sclerosis. The degree of neuromuscular block and rate of recovery were determined by measuring the amplitude of the first dorsal interosseus muscle action potential which was evoked periodically by trains of five stimuli (3 Hz each) to the ulnar nerve. In normal subjects, the first response to the train returned to its preinjection level after 20.0 +/- 8.5 minutes and the fifth response after 27.6 +/- 7.8 minutes. Multiple sclerosis patients showed a significantly delayed recovery of the first response (58 percent of patients) and of the fifth response (42 percent), suggesting a latent defect of neuromuscular transmission.


Multiple sclerosis: cellular and humoral immune responses to several viruses. One hundred and eight multiple sclerosis (MS) patients and 108 matched controls were studied for antibody levels and cellular immune responses to several viruses. There were significant increases in the mean titers of complement fixation (CF) or hemagglutination inhibition (HI), and complement-mediated cytotoxicity (CMC) tests for measles antibodies in MS patients; there was no increase in antibody titers to herpesviruses 1 and 2, or cytomegalovirus (CMV). The direct migration inhibition (DMI) tests showed no difference between MS patients and controls for measles, CMV, herpesviruses 1 and 2, or vaccinia virus antigens. Lymphocyte-mediated cytotoxicity (LMC) tests showed no difference between patients and controls, using cultures infected with measles and CMV viruses. In a study of stimulation or blocking of the LMC response by serum or cerebrospinal fluid (CSF), no effect was found. Therefore, increased levels of measles antibody in serum were again  demonstrated in  MS patients, but  there  was  no  difference  in  these  patients' cellular immunity to measles virus versus that of the controls, and there was no abnormality of cellular immunity against the other viruses tested.


The immunological response to intact and dissociated blue-tongue virus in mice. Antigenic fractions of bluetongue virus were separated by ultracentrifugation in Tris-buffered CsCl gradients at pH 6, 7 or 8 and the bluetongue virus polypeptide composition of the bands isolated from these gradeints was monitored by polyacrylamide gel slab electrophoresis. The immunological response to these fractions in mice was determined by a haemolytic plaque-forming cell assay, using sheep erythrocytes onto which intact bluetongue virus was adsorbed as lytic indicator cells. Isolated outer layer bluetongue virus polypeptide 2, from gradients at pH 6, and polypeptides 2 and 5, from gradients at pH 7, produced a strong primary IgM plaque-forming cell response. The subviral particles of density 1, 39 g.cm-3 and the bluetongue virus core particles of density 1,42 g.cm-3 also stimulated an IgM response at least as strong as that to intact bluetongue virus of density 1,38 g.cm-3. The isolated bluetongue virus fractions therefore appear to maintain their immunogenic integrity as effectively as those of intact bluetongue virus. The pattern of the immune response to bluetongue virus type 4 is similar to that of type 10.


Diarrhoea in pigs induced by rotavirus.

Diarrhoea in pigs induced by rotavirus. Electron microscope examination of the faeces of scouring pigs revealed virus particles which were morphologically indistinguishable from rotavirus (reo-like), a virus associated with diarrhoea in neonatal pigs (Lecce, King & Mock, 1976). This is the first record of the virus in the Republic of South Africa.


Preoperative facial paralysis in malignant parotid tumours. Preoperative paralysis of the facial nerve was found in 145 of 1,029 patients with malignant parotid tumours (14%) treated at nine university clinics in Scandinavia. The incidence of facial paralysis varied between the different clinics. A parellelism between the incidence of the facial paralysis and the impairment of the prognosis of the different tumour types is shown. The presence of preoperative facial nerve paralysis in malignant parotid tumours implies a very poor prognosis but the situation is not as hopeless as has been suggested and therefore one must rely on very radical surgery.


Isolation of blood and intracellular forms of Trypansoma cruzi from rats and other rodents and preliminary studies of their metabolism. Isolation of blood and intracellular forms of Trypanosoma cruzi was made mainly from rats (90-110 g) which had received 580 rad of whole-body gamma-irradiation not more than 24 h before subcutaneous inoculation with 10(7) trypomastigotes of the Sonya strain of T. cruzi. Unirradiated chinchillas (250-350 g) were, however, used for some experiments. Blood forms were isolated using a technique involving differential centrifugation to remove most of the erythrocytes and DEAE-cellulose chromatography to remove the remaining blood cells. Overall recoveries were usually in the range 30-70%. Parasites were mainly (approximately 98%) broad forms and were motile, metabolically active (as judged by respiratory and radio-tracer incorporation studies) and had lost none of their infectivity for mice. Intracellular forms were isolated from hind-limb muscle tissue. This was disrupted in an MSE tissue homogenizer and the homogenate incubated with DNase, collagenase and trypsin. Parasites, contaminated only by a few blood cells, were then obtained by differential centrifugation. For purer preparations, a terminal sucrose gradient step was used. Recoveries ranged between 40 and 70%. About 1-3% of the parasites isolated were epimastigotes and trypomastigotes; the remainder are probably best collectively termed 'amastigotes', though they were pointed and most had a short, free flagellum. They were undamaged as judged by light and electron microscopy and metabolically active as judged by respiratory and radio-tracer incorporation studies. However, the infectivity for mice of both these purified preparations and the initial cell homogenates could be accounted for by the epimastigotes and trypomastigotes present in them. Preliminary biochemical studies with isolated parasites have shown that blood, intracellular and culture forms of T. cruzi have a respiratory system which is in part sensitive to CN- and that all forms synthesize nucleic acids and proteins when incubated in vitro. There appears, however, to be a lack of DNA synthesis in blood stages, and thus it is not surprising that these forms do not divide.


Replication of Mouse Sarcoma Virus (Moloney) and its helper in a human heteroploid cell line of malignant origin. A human heteroploid cell line of malignant origin (J111) was infected with Mouse Moloney Sarcoma Virus. [MSV-M(MLV)]. The replication of the virus was demonstrated by different methods. After a few passages of infected cultures, the transforming component of MSV-M(MLV) disappeared from the supernatant fluids and cells. The non transforming helper component continued to be produced. The relatively low amount of helper virus released by the cells suggests a control of its replication in the J111 cellular system.


[Metyrapone test and plasma hormone estimations (author's transl)]. The usual metyrapone test based on the determination of the urinary steroid levels after ingestion of the product is relatively difficult to use because of its duration. In order to simplify this test, a comparative study was carried out with thirty subjects, 24 of whom were normal, by measuring their plasma hormonal levels by radioimmunoassay for ACTH, cortisol and aldosterone and by radiocompetition after extraction by carbon tetrachloride for the 11-desoxycortisol (S compound). The results show a parallel between the variation of the urinary steroid levels and the variations of plasma hormone levels during the first 24 hours of the test. Therefore, measuring the levels of ACTH, S compound and cortisol in the plasma before the ingestion of metyrapone and 24 hours afterwards seems simpler to carry out and accurate enough to test the integrity of the feed-back system between hypothalamus, pituitary and adrenal glands.


Cae I: an endonuclease isolated from the African green monkey with properties indicating site-specific cleavage of homologous and heterologous mammalian DNA. Component alpha DNA is a highly repetitive sequence that comprises nearly a quarter of the African green monkey (Cercopithecus aethiops) genome. A previous microbial restriction enzyme analysis showed that the repeat structure of component alpha DNA is based upon a monomeric unit of 176 +/- 4 base-pairs. An endonuclease, provisionally termed Case I, has been isolated from African green monkey testes that cleaves component alpha DNA into multimeric segments based upon the same repeat periodicity as that revealed by microbial restriction enzymes. The primary sites of Cae I cleavage in the component alpha sequence appear to be 120 +/- 6 base-pairs distant from the Hind III sites and 73 +/- 6 base-pairs distant from the Eco RI* sites. Cae I has been partially characterized with special reference to the effects of ATP and S-adenosylmethionine on the cleavage of component alpha DNA. Cae I may be a member of a class of similar site-specific nucleases present in mammalian cells. Cae I also cleaves mouse satellite DNA into a multimeric series of discrete segments: the periodicity of this series is shorter than that revealed by Eco RII retriction analysis of mouse satellite DNA.


The reactivity of phosphomono-and phosphodiester groups in oligonucleotides. The rate constants were estimated by phosphorus NMR spectroscopy for the reactions of alcohols (Tr-dT, 2-cyanoethanol) in pyridine with the main types of the reactive phosphorylating intermediates formed by treatment of pdT-Ac, pdTpdT-Ac, Tr-dTpdT-Ac, Tr-dTpdTpdT-Ac with 2, 4, 6-triisopropylbenzene-sulfonyl chloride (TPS): 1) B type derivatives with phosphomono ester (PME) group converted to a phosphoryl pyridinium residue; 2) C type derivatives with PME and phosphodiester (PDE) groups converted to trisubstituted pyrophosphate; 3) D type derivatives with PDE groups converted to tetrasubstituted pyrophosphate. The two latter types are partially present as cyclic intramolecular pyrophosphates Ci and Di. The reactivity of the intermediates decrease in the series B greater than Ci approximately Di greater than C approximately D. The Ci derivative of pdTpdT-Ac when obtained in dimethylformamide was found to be rather stable to hydrolysis and could be separated from the other dinucleotide derivatives by ion-exchange chromatography. The Arrhenius parameters of all steps of the conversion of PME group of pdT-Ac to B derivative and of the reaction of TPS with PDE group of dinucleoside phosphate Tr-dTpdT-Ac were measured.


Cleavage of Epstein-Barr virus DNA by restriction endonucleases EcoRI, HindIII and BamI. The cleavage of the DNAs of the B95-8 and P3HR-1 virus strains of Epstein-Barr virus by the restriction endonucleases EcoRI, HindIII and BamI was investigated using a new technique for quantitative evaluation of the fluorescence of ethidium stained DNA fragments separated on agarose gels. The results obtained with B95-8 DNA showed that in addition to the limited repetitions of nucleotide sequences observed in the EcoRI and HindIII cleavage patterns, the molecule contained a BamI fragment with a molecular mass of 2.0 megadaltons which was present in a total of about 11 copies and localized to a limited part of the DNA molecule. The same sequences were also present in the P3HR-1 DNA albeit in a lower molar ratio. P3HR-1 DNA yielded restriction enzyme cleavage patterns suggesting DNA sequence heterogeneity of P3HR-1 virus. No fragment was present in more than about 4 copies per molecule of P3HR-1 DNA. Comparison of the restriction enzyme cleavage patterns of P3HR-1 and B95-8 DNA revealed a high degree of structural homology emphasized by nucleic acid hybridization experiments with EBV complementary RNA synthesized in vitro.


Photochemical crosslinking of transcription complexes with psoralen. I. Covalent attachment of in vitro SV40 nascent RNA to its double-stranded DNA template. 14C-labeled SV40 DNA has been transcribed with E. coli RNA polymerase using 3H-labeled ribonucleotide triphosphates as precursors. The resulting transcription complexes were then photochemically crosslinked with the psoralen derivative, 4'-aminomethyl-4,5', 8-trimethylpsoralen (AMT), at 37 degrees C and analyzed in SDS-sucrose gradients. It was found that the photochemical crosslinking procedure caused the nascent RNA chains to co-sediment with their double-stranded (helical) SV40 templates in the denaturing sucrose gradient. This result and several control experiments suggest that covalent linkages have formed between nascent RNA and helical DNA after the photochemical reaction. The crosslinking phenomenon was observed to be independent of the superhelical state of the DNA used as the template. Prior addition of EDTA to stop the transcription is not required for successful crosslinkage.


Discharge of the premature infant.

Discharge of the premature infant. Appropriate interventions are initiated smoothly if the neonatal staff is familiarized with the new concepts of infant care and their empirical basis. It is also important that the nurses be present at all conferences to facilitate more meaningful interaction with the mother and her infant. If this is done, then it is easier to evaluate the needs of the mother and how much teaching support is necessary. The more involved the nurse becomes with the family, the more rewarding her experience will be, and she will be eager to get involved with the next family.


Common methionine-tryptic peptides near the amino-terminal end of primate papovavirus tumor antigens. The tumor antigens directed by human papovaviruses BK and JC and the monkey papovavirus simian virus 40 have two methionine-containing tryptic peptides in common. These peptides are constituents of the small forms of papovavirus tumor antigen (17,000 daltons) which are present in lytically infected and transformed cells and which are believed to share some amino acid sequences with the amino-terminal portion of the larger tumor antigen species (97,000 daltons). In addition to the two peptides, which are present in all three papovavirus tumor antigens, the larger forms of the tumor antigens specified by simian virus 40 and BK virus share four other methionine-containing tryptic peptides, two of which are also present in the smaller (17,000 daltons) species of antigen. The occurrence of common peptides at the amino-terminal portion of tumor antigens of primate papovaviruses suggests that these conserved regions may play a fundamental role in the function of these proteins and in the propagation of these viruses in nature. The tryptic peptides of the small forms of papovavirus tumor antigen were examined and compared to those present in the large species. Out of a total of nine and ten methionine-containing peptides in the 17,000-dalton tumor antigens of simian virus 40 and BK virus, seven and nine peptides, respectively, are constituents of the corresponding larger (97,000 daltons) forms of the antigen.


Translational control by hemin is due to binding to cyclic AMP-dependent protein kinase. Our previous work [Proc. Natl, Acad. Sci. USA (1977) 74, 1463-1467, 3326-3329] is consistent with the view that (a) the hemin-controlled inhibitor of protein synthesis in reticulocyte lysates (active eIF-2 kinase) is formed by phosphorylation of proinhibitor (inactive eIF-2 kinase) catalyzed by cyclic AMP-dependent protein kinase (ATP-protein phosphotransferase; EC 2.7.1.37), and (b) hemin prevents this conversion by blocking the interaction of cyclic AMP with the kinase's regulation subunit, thereby rendering the enzyme inactive. We now show that hemin blocks cyclic AMP binding because it itself binds specifically to the regulatory subunit. This binding is noncompetitive with respect to cyclic AMP. Whereas unlabeled hemin can displace bound [3H]hemin as well as cyclic [3H]AMP, unlabeled cyclic AMP can displace bound cyclic [3H]AMP but not [3H]hemin. This suggests that cyclic AMP and hemin bind to different sites on the protein and that hemin binding affects cyclic AMP binding in an allosteric manner.


Fluorescent photoaffinity labeling: adenosine 3',5'-cyclic monophosphate receptor sites. An approach to the study of protein receptor sites in protein mixtures or supramolecular assemblies by using fluorescence spectroscopy is described. This approach, fluorescent photoaffinity labeling, combines the merits of photoaffinity labeling to attain site-directed reactivity with the probing power of fluorescent ligands. A fluorescent photoaffinity label for cyclic AMP receptor sites of cyclic AMP-dependent protein kinases was synthesized in both unlabeled and radioactive forms. The probe, 8-azido-1,N(6)-ethenoadenosine 3',5'-cyclic monophosphate, mimics cyclic AMP in its ability to stimulate the phosphotransferase activity of the protein kinases and strongly competes with cyclic AMP for its binding sites in all preparations so far tested. Photolysis, after equilibration of protein kinase and 8-azido-1,N(6)-ethenoadenosine 3',5'-cyclic monophosphate in the dark, effects binding of the intermediate nitrene irreversibly and specifically to the cyclic AMP sites with the development of fluorescence. Excess reagent and low molecular weight photolytic products are removable by dialysis. Studies of a crude beef heart preparation containing cyclic AMP-dependent protein kinase suggest that the cyclic AMP binding sites are hydrophobic in nature and strongly immobilize the adenine moiety of the cyclic nucleotide.


Amphibian oocyte maturation and protein synthesis: related inhibition by cyclic AMP, theophylline, and papaverine. Two inhibitors of cyclic AMP phosphodiesterase (3':5'-cyclic-AMP 5'-nucleotidohydrolase, EC 3.1.4.17), theophylline and papaverine, inhibit the maturation of Xenopus laevis oocytes induced by four different stimuli: human chorionic gonadotropin, progesterone, testosterone, and lanthanum ions. Addition of 1 mM cyclic AMP to the medium delays maturation by approximately 2 hr. Papaverine, theophylline, and cyclic AMP inhibit amino acid incorporation into oocyte proteins by 50% or more but do not inhibit amino acid uptake. The capacity of theophylline to block maturation and protein synthesis is reversed in a parallel fashion by addition of 1-5 mM calcium ion to the medium. Addition of papaverine, theophylline, and cycloheximide to oocytes at different times after hormonal treatment shows that the step sensitive to blockage by the three drugs is coincident and precedes germinal vesicle breakdown by about 1.5 hr. Theophylline and papaverine do not increase endogenous cyclic AMP levels in oocytes but do block the decrease of cyclic AMP levels observed 3 hr after progesterone treatment. Both drugs inhibit oocyte cyclic AMP phosphodiesterase measured in vivo and severely inhibit the stimulus of calcium uptake caused by progesterone and human chorionic gonadotropin. These results suggest that cyclic AMP, theophylline, and papaverine may block oocyte maturation by inhibiting protein synthesis, possibly via a cyclic AMP-dependent protein kinase as shown in reticulocytes [Datta, A., De Haro, C., Sierra, J. & Ochoa, S. (1977) Proc. Natl. Acad. Sci., USA 74, 1463-1467].


Detection and kinetic behavior of preproinsulin in pancreatic islets. Newly synthesized rat islet proteins have been analyzed by polyacrylamide slab gel electrophoresis and fluorography. A minor component having an apparent molecular weight of 11,100 was identified as preproinsulin by the sensitivity of its synthesis to glucose, the pattern of NH2-terminal leucine residues, and the rapidity of its appearance and disappearance during incubation of islets or islet cell tumors. A small amount of labeled peptide material which may represent the excised NH2-terminal extension of preproinsulin or its fragment was also detected. The kinetics of formation and processing of the preproinsulin fraction were complex, consisting of a rapidly turning over component having a half-life of about 1 min and a slower minor fraction that may have bypassed the normal cleavage process. The electrophoretic resolution of the preproinsulin and proinsulin fractions into two bands each is consistent with the presence of two closely related gene products in rat islets rather than intermediate stages in the processing of these peptides.


Spliced early mRNAs of simian virus 40.

Spliced early mRNAs of simian virus 40. Biochemical methods are presented for determining the structure of spliced RNAs present in cells at low concentrations. Two cytoplasmic spliced viral RNAs were detected in CV-1 cells during the early phase of simian virus 40 (SV40) infection. One is 2200 nucleotides in length and is composed of two parts, 330 and 1900 nucleotides, mapping from approximately 0.67 to approximately 0.60 and from approximately 0.54 to approximately 0.14, respectively, on the standard viral map. The other is 2500 nucleotides long and also is composed of two parts, 630 and 1900 nucleotides mapping from approximately 0.67 to approximately 0.54 and from approximately 0.54 to approximately 0.14, respectively. Correlation of the structure of these mRNAs with the structure of the early SV40 proteins, small T antigen (17,000 daltons) and large T antigen (90,000 daltons), determined by others suggests that: (i) translation of the 2500-nucleotide mRNA yields small T antigen; (ii) translation of the 2200-nucleotide mRNA proceeds through the splice point in the RNA to produce large T antigen (and thus large T antigen is encoded in two separate regions of the viral genome); and (iii) the DNA sequences between approximately 0.67 and approximately 0.60 present in both mRNAs are translated in the same reading frame in both mRNAs to yield two separate gene products that have the same NH(2)-terminal sequence. Therefore, expression of the early SV40 genes is partially controlled at the level of splicing of RNAs.


Biological activity of purified simian virus 40 T antigen proteins. Proteins related to simian virus 40 (SV40) T antigen uere isolated from cells infected with adenovirus 2/SV40 hybrids Ad2+D2 and Ad2+ND1 dp2 as well as from a line of human cells (SV80) transformed by SV40. The 96,000- and 107,000-dalton proteins of SV80 and Ad2+D2, after injection into the cytoplasm of cultured cells, rapidly accumulate in the nuclei, where they remain antigenically reactive for at least 20 hr and trigger DNA synthesis in quiescent cells. By contrast, the 23,000-dalton protein coded by Ad2+ND1 dp2 does not stimulate cellular DNA synthesis. However, all three purified proteins are able to provide helper function for the growth of adenovirus 2 in monkey cells. Thus, purified SV40 T antigen and proteins that share sequences with it retain the ability to carry out at least two functions associated with the product of the A gene of SV40.


Evidence for splicing of avian sarcoma virus 5'-terminal genomic sequences into viral-specific RNA in infected cells. The 5'-terminal nucleotide sequences of the avian sarcoma virus (ASV) genome are transcribed by the reverse transcriptase in vitro into a DNA transcript that represents the entire distance ( approximately 100 nucleotides) between the tRNA(Trp) primer molecule and the 5' terminus. We have used these DNA(100) transcripts in hybridization reactions with ASV-specific RNA from infected avian cells and find nucleotide sequences complementary to these transcripts on all of the various size classes of viral mRNA identified. Similar hybridization results were obtained with a specific DNA transcript complementary to viral genomic nucleotide sequences between the tRNA(Trp) primer molecule and up to, but not including, the terminal redundant sequences (DNA(70)), indicating that the observed hybridization of DNA(100) to all size classes of viral RNA in infected cells did not reflect hybridization of DNA(100) to the terminal redundant sequences at the 3' end of the viral genome. Escherichia coli RNase H hydrolysis of RNA.DNA hybrids consisting of genomic 35S RNA obtained from virus and DNA(100) transcripts indicated that viral genomic sequences complementary to these DNA transcripts were not present at sites distal to the ends of the RNA genome and therefore not adjacent to the corresponding gene sequences representing the various species of viral mRNA from infected cells. These studies suggest that the 5'-terminal genomic nucleotide sequences, or a portion thereof, are somehow added or "spliced" onto each ASV-specific mRNA species in infected cells either during or after transcription of proviral DNA for some as yet undetermined purpose.


Regulation of acetylcholine release from neuroblastoma x glioma hybrid cells. Neuroblastoma x glioma NG108-15 hybrid cells exposed to N6, O2'-dibutyryladenosine 3':5'-cyclic monophosphate for several days release [3H]acetylcholine in response to serotonin, prostaglandin F2alpha, KCl, or veratridine. NG108-15 cells grown in the absence of dibutyrul cyclic AMP do not respond to an excitatory stimulus by releasing [3H]acetylcholine but can be shifted to a responsive state by treatment with dibutyryl cyclic AMP. Thus, the reactions that are required for acetylcholine release can be regulated in NG108-15 cells, thereby regulating the ability of cells to form synapses and the efficiency of synaptic communication.


Mechanism of the rapid antigonadotropic action of prostaglandins in cultured luteal cells. A reproducible method for dissociation and culture of rat luteal cells is described. The concentration of LH required to produce half-maximal stimulation of progesterone secretion was 50 ng/ml. The effects of prostaglandin E(2) (PGE(2)) and prostaglandin F(2alpha) (PGF(2alpha)) on basal and luteinizing hormone (LH)-stimulated progesterone production were examined. Both prostaglandins stimulated basal progesterone production but PGE(2) was about twice as active, showing a 2-fold maximal stimulation at 0.75 muM. When either prostaglandin was incubated simultaneously with LH, a dose-dependent inhibition of progesterone secretion occurred; PGF(2alpha) was 4 times more active than PGE(2), showing 50% inhibition at a concentration of 40 x nM. Thus, both prostaglandins are more active as antagonists than as agonists of LH with respect to progesterone secretion. PGF(2alpha) also inhibited LH-stimulated adenylate cyclase activity and cyclic AMP accumulation. The block in progesterone secretion was reversed by addition of dibutyryl cyclic AMP (1 mM) but not by theophylline (5 mM) alone. These data and the finding that PGF(2alpha) did not affect the specific binding activity of the LH receptor in intact luteal cells indicate that the rapid action of prostaglandins in luteal cells is due to a block of LH-dependent production of cyclic AMP which results in a decrease in progesterone secretion.