Potentiation of postjunctional cholinergic sensitivity of rat diaphragm muscle by high-energy-phosphate adenine nucleotides. The cholinergic sensitivity of rat diaphragm muscle, me-sured as the magnitude of depolarization responses to repetitive, iontophoretic pulses of acetylcholine (ACh) onto neuromuscular endplates, is increased by addition of ATP to the perfusion medium. Depolarization responses begin to increase within the first min after addition of 10 mM ATP and plateau at 60% above control levels (mean value) after 4 to 6 min. Neither the magnitude nor the time course of the potentiations corresponds to changes in resting potential or membrane resistance. Other nucleotides are equally or less effective at the same concentration: ATP=ADP greater than UTP greater than AMP=GTP (=no added nucleotide control) The duration of the individual ACh responses does not increase during continuous exposure to the active nucleotides for up to 15 min except when the muscle is pretreated with eserine. Mild enzymatic predigestion of the muscle with collagenase and then protease, increasing the availability of the postjunctional membrane to bath-applied drugs, decreases the variability and increases the magnitude of the potentiation to a given dose of ATP. The dose-response curve for ATP is then more than half-maximal at 1 mM and the ranking of the other nucleotides relative to ATP is the same as without predigestion. There is an optimum Ca++ concentration for the potentiation between zero and 2 mM: potentiation is enhanced in Ca++ -free medium, partially blocked in twice-normal Ca++ medium, and totally blocked in Ca++ -free medium 10 min after a 5 min exposure to 2.5 mM EGTA. The similar Ca++ dependence of ACh receptor activation in the absence of added nucleotide suggests that ATP directly facilitates receptor activation by ACh. This facilitory action could be one of the physiological roles for the ATP released from stimulated phrenic nerve.


Modulation of postjunctional cholinergic sensitivity of rat diaphragm muscle by cyclic adenosine monophosphate. Addition of 2.5 mM cyclic adenosine monophosphate (cAMP) to the solution bathing a rat diaphragm muscle alters the magnitude of depolarization responses to iontophoretic pulses of acetylcholine (ACh) at neuromuscular endplates. Alterations are repeatable with small variability on a given preparation for initial and repeat experiments on both hemidiaphragms, but are different on each preparation. Five min after addition of the nucleotide solution, increases (potentiations) of up to 30% above control levels and decreases (attenuations) to 50% below control levels are observed. The effects on sensitivity to ACh of dibutyryl cAMP (1.25 mM), monobutyryl cAMP (0.25 mM), and cAMP (2.5 mM) in Ca++ -free solution are a function of whether the experiment is an initial one on that preparation or a repeat experiment after 10 or more minutes of perfusion flow. In all three cases, initial exposure attenuates sensitivity (means at 5 min: --30, --10, and --20%, respectively) and repeat exposure potentiates sensitivity (mean: 20% at 5 min, 15% at 5 min, and 10% at 2 min respectively). A concentration of dibutyryl cAMP (0.25 mM) which is without effect on sensitivity alone, produces a large, transient potentiation (mean: 45% at 1 min) in conjunction with 0.5 mM theophylline. A decrease in the rate of desensitization is observed during exposure to 0.25 mM cAMP. The results are interpreted in terms of a physiological mechanism whereby receptor activity at the postjunctional membrane is modulated by cAMP formed from prejunctionally released ATP.


Form and distribution of actin and myosin in non-muscle cells: a study using cultured chick embryo fibroblasts. Attempting to throw light on the mechanical basis of movement of non-muscle (cf. muscle) cells, the present work aims to determine the form and distribution of actin and myosin in chick embryo fibroblasts. These cells were cultured on formvar, fixed in glutaraldehyde then osmium tetroxide vapours, dehydrated, critical-point dried and examined, in toto, in the electron microscope (EM). Stereoscopic pairs of micrographs were studied to define more exactly the form and distribution of cytoplasmic filaments topographically associated with deformations of the cell surface and with organelle movements through the cytoplasm. Permeating the cytoplasm, interconnecting long and short filaments closely surrounded all organelles, linked with microtubules and polyribosomes and joined to the plasma membrane. These filaments, which varied greatly in width (2-13 nm) were closely associated with large numbers of 'comma-shaped' globoid bodies of approximately 15 nm diameter. Attempting to establish the identity, form and distribution of cytoplasmic myosin, cultured cells were extracted with a cold (4 degrees C) glycerol/pyrophosphate solution for 24 h before being fixed and critical-point dried. EM examination of these cells revealed a residual three-dimensional network of branching and anastomosing 4-13 nm diameter smooth filaments, devoid of fine (2 nm) filaments and globoid bodies. Examination of fixed, critical-point dried, skeletal muscle heavy meromyosin showed globoid structures similar in form and size to the globoid bodies found in cultures fibroblasts. Similarly fixed and critical-point dried paracrystals of actin, polymerized in the presence of Mg2+, appeared as branching interconnecting filaments which, in form and dimensions, resembled the network filaments observed in pyrophosphate-extracted cells. It is concluded that the pyrophosphate-extractable globoid bodies found in cultured fibroblasts represent monomers of myosin, that the broader filaments to which these attach represent actin in Mg2+ paracrystalline form and that the various subcellular movements are brought about by interactions between the two, analogous to those occurring in muscle cells.


Evolution of DNA structure: direction, mechanism, rate. On the basis of the results of an analysis of frequencies of pyrimidine oligonucleotides, the degree of pyrimidine clustering of DNA in species from different taxa has been determined. A tendency for an increase in the index of clustering of DNA was revealed in the sequence: invertebrates, fishes, amphibians, reptiles, birds, mammals. A mechanism is postulated, according to which the increase in the degree of clustering of DNA d-ring the evolution may be associated with the accumulation of mutations, Purine equalibrium Pyrimidine transversions, resulting in a selective enrichment of one of the chains of DNA with pyrimidines and the other- with purines, i.e. in an increase in the degree of purine-pyrimidine imbalance (asymmetry) of DNA complementary chains. This mechanism of DNA evolution is supported by the presence of positive correlation between the degree of clustering and the degree of the chain asymmetry of natural DNAs, as well as the character of the amino acid substitutions in cytochromes c in different species. The progressive evolution of different groups of organisms on the whole may have been accompanied by an acceleration of the rates of evolution of the DNA structure. On the basis of the amino acid sequence of cytochromes c in different species the degree of clustering and the degree of the chain asymmetry of the corresponding structural genes of DNA was found to have a general tendency towards an increase in the following order: invertebrates, fishes, amphibians, reptiles, birds, mammals. Thus, evolution of cytochrome c cistron is a vector process based on a selection of mutations which, on the one hand, are neurtral to protein, and, on the other hand, result in the sense chain of DNA being enriched with pyrimidines and the nonsense one (and the corresponding mRNA)- with purines. Hence, it is the polynucleotide template rather than protein, that must have been the "object of selection". The frequency of substitutions in cytochromes c cistron for vertebrates is 1.56x13(-9) per nucleotide per year. It is believed that the evolutionary modification of the DNA structure may be associated with an increase in the interference resistance of the translation, i.e. with selection for codons of highest readout stability.


Comparative diagnostic aspects of herpes simplex virus tumor-associated antigens. Sera from cancer patients and healthy individuals, obtained from two independent sources, were examined for their abilities to react with herpes simplex virus-associated tumor antigens, AG-4 and NVA-TAA (nonvirion antigen-tumor-associated antigen). Both antigens were prepared by infection of HEp-2 cells with herpes simplex virus type 2, and all antigen-antibody interactions were measured by the micro-complement fixation test. Of sera from 16 patients with cancer of the uterine cervix, 81% (P less than 0.01) reacted with NVA-TAA, whereas 78% (P less than 0.001) of 18 sera examined reacted with AG-4. These values differed significantly from those for normal sera, of which 14% reacted with NVA-TAA and 13% with AG-4. Of sera for 8 patients with squamous cell carcinoma of head and neck or vulva, 75% (P less than 0.02) reacted with NVA-TAA, whereas 63% (P less than 0.05) reacted with AG-4. As a group, other cancers (including adenocarcinoma of lung, breast, ovary, and cervix; liposarcoma; sarcoma; melanoma; and carcinoma of the endometrium) did not differ significantly from controls in reactive patterns with AG-4 or NVA-TAA. These studies partly supported the reported preferential reactivity of AG-4 and NVA-TAA with sera of patients with squamous cell carcinoma, especially of the uterine cervix.


Pathogenesis of malignant histiocytic lymphoma induced by silica in a colony of specificpathogen-free Wistar rats. The occurrence of malignant histiocytic lymphomas (MHLT) after an intrapleural inoculation of silica and the presence of a hyperplastic or granulomatous reaction to the silica were confirmed in cesarean-derived barrier-maintained inbred W rats. A specific hyperplastic reaction in the silica granulomas preceded the appearance of MHLT. Hyperplastic changes were observed when the absolute monocyte count at weeks 25-30 was slightly higher than that seen in the controls. Silica injected intrapleurally penetrated the thymus tissue but the carbon and coal dusts and the silica, which was administered by other routes, did not involve the thymus. Also, silica was the sole dust, when introduced into the coelomic cavities that produced MHLT, which grew rapidly on entering the thymus. Rats that died with MHLT had widespread dissemination with malignant foci situated most consistently above the diaphragm.


Autogenous humoral immunity to baboon xenotropic endogenous type C virus. A radioimmune precipitation assay was used to evaluate the humoral immune response of baboons (Papio cynocephalus) to their endogenous, xenotropic, type C (M7) virus. Using 125I or [3H]leucine-labeled intact virus preparation, we demonstrated that baboons possess naturally occurring virus-binding antibodies to this virus. Antibody appeared directed against envelope and not internal viral proteins. Immunoglobulins responsible for virus-binding were predominant in the IgG and IgA serum fractions, whereas IgM participated to a minor degree. The biologic activities of these immunoglobulins were investigated by means of a focus reduction assay for virus neutralization with a murine sarcoma virus (M7) pseudotype. We determined that, despite the presence of considerable virus binding, none of the normal baboon sera tested possessed neutralizing activity for M7 virus.


Type C virus expression in lymphoma-paralysis-prone wild mice. Wild mice trapped near Lake Casitas (LC) in southern California showed a high prevalence of infectious type C virus in the liver, spleen, and thymus within the first few weeks of life. By young adulthood about 80% of LC mice (including their genital tissues) were infected. Virus isolates from these mice cause lymphoma and lower limb paralysis under both natural and experimental conditions. Mice destined to develop paralysis showed higher levels of serum gs antigen early in life, whereas mice destined to develop lymphoma or remain free of these diseases could not be distinguished by this test. The individual variation in virus expression suggested that differences in virus type or in the immune or other host defense mechanisms greatly influenced susceptibility or resistance to indigenous type C virus-caused disease in LC wild mice.


Developmental genetics of neuroblastoma. Case reports of neuroblastoma revealed that some individuals are genetically predisposed and that this genetic predisposition may have other consequences. According to a mutation model, two classes of individuals could acquire neuroblastoma. One (prezygotic) was a rare class that carried a dominant gene imparting high risk of the tumor. The other (postzygotic) comprised all other individuals, each at low risk. The model related tumor incidence to germinal and somatic mutation rates and thereby carried implications for environmental modification of tumorigenesis and demographic variation in incidence. Case reports also revealed associations of neuroblastoma with congenital defects and a susceptibility to second tumors. Analogy with retinoblastoma and Wilms' tumor of the kidney suggested that these associations could result from action of a neuroblastoma gene or from chromosomal aberration. One known dominantly inherited condition, von Recklinghausen's disease, could dispose to neuroblastoma and create some associations. According to the two-mutation model, neuroblastoma may have been a single recessive gene disorder at the level of the cell. The phenomena of aganglionosis, neuroblastoma in situ, maturation of neuroblastoma to ganglioneuroma, and spontaneous regression suggested that such a neuroblastoma gene interfered with normal developmental processes. The specificities of this gene and of those for von Recklinghausen's disease and pheochromocytoma suggested that the functiof a membrane macromolecule.


Neuroblastoma, Wilms' tumor, and cytomegalovirus. The possibility that neuroblastomas and Wilms' tumors are induced by cytomegalovirus (CMV) was raised and discussed. A large percentage of patients with these tumors have complement-fixing antibodies against CMV. A hypothesis was presented.


A rationale for the treatment of metastatic neuroblastoma. In developing a chemotherapeutic program for children with disseminated neuroblastoma, we established three human neuroblastoma lines in cell culture to study the effects of dibutyryl cyclic AMP, papaverine, 5-trifluoromethyl-2'-deoxyuridine, and cyclophosphamide on cell growth, biochemical behavior, and morphology. Based upon our studies, a clinical treatment program was designed. We have treated 15 patients with disseminated neuroblastoma and have established the optimum dose range and sequence of these drugs. Early results were promising; plans for continuation of clinical and experimental studies were discussed.


Methylation of high-molecular-weight subunit RNA of feline leukemia virus. The high-molecular-weight subunit RNA of feline leukemia virus (Rickard strain) (FeLV-R) was analyzed for the presence of methyl groups. After purification of native 50-60S FeLV-R RNA on nondenaturing aqueous sucrose density gradients. FeLV-R 28S subunit RNA, doubly labeled with [14C]uridine and [methyl-3H]methionine, was isolated by centrifugation through denaturing sucrose density gradients in dimethyl sulfoxide. As calculated from their respective 3H/14C ratios. FeLV-R 28S RNA was methylated to the same degree as host cell poly(A)+ mRNA. When the 28S FeLV-R RNA was hydrolyzed to completion with RNase T2 or alkali, all of the methyl-3H chromatographed with mononucleotides on Pellionex-WAX, a weak anion exchanger. The methyl-labeled material co-chromatographed with 6-methyladenosine if the mononucleotide fraction obtained by Pellionex-WAX chromatography was hydrolyzed to nucleosides by bacterial alkaline phosphatase or with 6-methyladenine if purine bases were released from the mononucleotides by acid hydrolysis. In another experiment in which FeLV-R 28S RNA uniformly labeled with 32P was hydrolyzed and then analyzed by Pellionex-WAX chromatography, all of the 32P label again co-chromatographed with mononucleotides. Thus FeLV-R 28S RNA does not appear to contain a 5' structure, either methylated or nonmethylated similar to those recently reported for cellular and some animal virus mRNA's.


Polyoma genome in hamster BHK-21-C13 cells: integration into cellular DNA and induction of the viral replication. When grown at 39.5 degrees C, BHK-21 C-13 cells transformed by A gene mutants of polyoma virus contain viral sequences that are predominantly associated with cellular DNA pelleted in the Hirt lysis procedure. At this temperature, in cells that are inducible for viral DNA replication (Folk, 1973), the majority of the viral genomes are covalently joined with cellular DNA's containing repetitious sequences. Upon a shift to 31 degrees C, free viral genomes appear and are replicated. Coupled with the replication of the free viral genomes at 31 degrees C is an increase in the viral genomes associated with cellular DNA.


Synthesis of RNA by mutants of vesicular stomatitis virus (Indiana serotype) and the ability of wild-type VSV New Jersey to complement the VSV Indiana ts G I-114 transcription defect. The ability of certain vesicular stomatitis virus (VSV; Indiana serotype) temperature-sensitive (ts) mutants to synthesize intracellular viral complementary RNA (vcRNA) at permissive or nonpermissive temperatures for productive infections has been investigated. Mutants belonging to complementation groups II, III, and V synthesize RNA at nonpermissive temperature in amounts essentially equivalent to that obtained at permissive temperatures. Mutant ts G I-114 possesses a thermolabile transcriptase and does not synthesize vcRNA at 40 degrees C; however, mutants ts O I-5, O I-53, O I-78, and O I-80 possess thermostabile transcriptases that are capable of some vcRNA synthesis at 40 degrees C. All five group I mutants are defective in their secondary transcription ability at 40 degrees C. Wild-type VSV New Jersey virus is able to complement the transcription defect of ts G I-114 at 40 degrees C. This complementation is inhibited by puromycin, suggesting that a viral gene product of VSV New Jersey (e.g., its transcriptase or a transcriptase component) is involved. Mokola virus is not able to complement the ts G I-114 defect, although Mokola does synthesize vcRNA in infected cells (in  the presence or absence of cycloheximide).


Polyadenylate sequences of human rhinovirus and poliovirus RNA and cordycepin sensitivity of virus replication. The polyadenylate [poly(A)] content of the genome RNA of human rhinovirus type 14 (HRV-14) is nearly twice as large as that of the genome RNA of poliovirus type 2. The poly(A) content of viral RNA was determined to be the RNase-resistant fraction of 32P-labeled viral RNA extracted from purified virions. Polyacrylamide gel electrophoresis indicated that the poly(A) sequences of HRV-14 are more heterogenous and on an average larger than those of poliovirus RNA. On the basis of susceptibility to micrococcal polynucleotide phosphorylase the rhinovirus genome terminates in poly(A). Replication of both viruses is almost totally inhibited by cordycepin at 50 mug/ml. At lower concentrations, rhinovirus replication is more sensitive to cordycepin than poliovirus replication. Addition of cordycepin (75 mug/ml) to infected culture prior to or during viral RNA replication results in more or less complete inhibition of virus-specific RNA synthesis. The results do not indicate that cordycepin sensitivity of either virus is due to preferential inhibition of viral poly(A) synthesis by this antibiotic.


Sequence arrangement in herpes simplex virus type 1 DNA: identification of terminal fragments in restriction endonuclease digests and evidence for inversions in redundant and unique sequences. It has been proposed by Sheldrick and Berthelot (1974) that the terminal sequences of herpes simplex virus type 1 (HSV-1) DNA are repeated in an internal inverted form and that the inverted redundant sequences delimit and separate two unique sequences, S and L. In this study the sequence arrangement in HSV-1 DNA has been investigated with restriction endonuclease cleavage, end-labeling studies, and molecular hybridization experiments. The terminal fragments in digests with restriction endonucleases Hind III, Hpa-1, EcoRI and Bum were identified and shown to be consistent with the Sheldrick and Berthelot model. Inverted fragments which contain unique sequences as well as redundant sequences, and which the model predicts, were identified by DNA-DNA hybridization studies. Further cleavage of Bum fragments with Hpa-1 also revealed inversions of the terminal sequences that contained unique sequences. The results obtained showed that the unique sequences S and L are relatively inverted in different DNA molecules in the population, resulting in the presence of four related genomes with rearranged sequences in apparently equal amounts. The redundant sequences bounding S do not share complete sequence homology with those bounding L, but hybridization studies are presented which show that the terminal 0.3% of the genome is repeated in every redundant sequence.


Physical maps for Herpes simplex virus type 1 DNA for restriction endonucleases Hind III, Hpa-1, and X. bad. It has been proposed that the genome of herpes simplex virus type 1 (HSV-1) consists of two internal unique sequences, S and L, bounded by two sets of redundant sequences (P. Sheldrick and N. Berthelot, 1974). In this arrangement, terminal sequences (TRs and TRl) are repeated in an internal inverted form (IRs and IRl) and delimit S and L. Furthermore, a body of evidence has accumulated that suggests that S and L themselves are inverted, giving rise to four related forms of the HSV genome. In this study the ordering of restruction endonuclease fragments of HSV-1 DNA for physical maps has been studied using molecular hybridization techniques and the cleavage of isolated restriction endonuclease fragments with further restriction endonucleases. Physical maps for the fragments produced by Hind III, Hpa-1, and X. bad have been constructed for the four related forms of the HSV-1 genome. TRs and IRs were found to be between 3.5 x 10(6) and 4.5 x 10(6) daltons, TRl and IRl about 6 x 10(6) daltons, S about 8 x 10(6) to 9 x 10(6) daltons, and L about 6.8 x 10(6) daltons.


Generation of defective virus after infection of newborn rats with reovirus. When 2-day-old rats were inoculated subcutaneously with the R2 strain of reovirus type 3 or with a class B (352) or class C (447) temperature-sensitive (ts) mutant, 5 to 10% of the animals died from acute encephalitis within 12 days. Approximately half of the survivors recovered rapidly and grew normally, but the remainder became runted. Two phases of infection are distinguished in the animals: an acute phase during which infectious virus reaches a maximum titer in brain and other tissues by 10 days p.i. and thes runting of the rats and the slow disappearance of virus from their brains over a period of 2 months or so. Virus isolated from chronically infected brains generally retained the genetic character (ts or wild type) of the inoculated virus, but two exceptions to this are described. Defective virions lacking the L1 segment of the viral genome (L1 defectives) were generated in rat brains during the acute phase of infection. Defective virus was also generated during the chronic phase, but during this period defectives were found with multiple segments deleted from the genome in addition to L1 defectives. In another type of experiment defective virus exerted a marked protective effect when inoculated intracerebrally with R2 virus. In the absence of defectives all animals died, but in their presence 17 of 23 animals survived and 15 of 23 became runted and chronically infected. The formation and evolution of defective particles in the brains of these rats were similar to those found in rats chronically infected after subcutaneous inoculation of reovirus. We conclude that the formation of defective virus particles may play a role in the initiation and maintenance of chronic neutropic infections with reovirus.


5'-terminus of Moloney murine leukemia virus 35s RNA is m7G5' ppp5' GmpCp. The 5'-terminal sequence m(7)G(5') ppp(5') GmpCp was isolated from Moloney murine leukemia virus 35S RNA after digestion of (32)P-labeled RNA with RNases T1, T2, and A followed by pH 3.5 ionophoresis on DEAE paper.


RNA complementary to the genome of RNA tumor viruses in virions and virus-producing cells. Cells producing type C (avain sarcoma virus) or type B (mouse mammary tumor virus) RNA tumor viruses contain small amounts of RNA complementary to the viral genomes. The negative strands are complementary to at least 30 to 45% of the viral genomes and are found as RNA-RNA duplexes in the nucleus and cytoplasm of infected cells and in mature virions.


Simian virus 40 mutants carrying two temperature-sensitive mutations. Five temperature-sensitive mutants of simian virus 40 containing two temperature-sensitive mutations were isolated. The double mutant of the A and D complementation groups, like the D mutants, failed to complement by conventional complementation analysis and did not induce host DNA synthesis at 40 degrees C. However, under conditions that suppressed the D defect, the A:D double mutant expressed only the A defect. Thus, viral DNA replication dropped rapidly after this mutant was shifted from permissive to restrictive temperatures. The A:D double mutant failed to transfrom at the restrictive temperature when subconfluent Chinese hamster lung monolayers were used. Double mutants of A:B, A:C, and A:BC complementation groups, like their A parent, were defective in viral DNA replication, in the induction of host DNA synthesis and in the transformation of secondary Chinese hamster lung cells at the nonpermissive temperature.


Consequences of herpes simplex virus type 2 and human cell interaction at supraoptimal temperatures. The consequences of herpes simplex virus type 2 (HSV-2) and human embryonic fibroblast cell interaction at different temperatures (37, 40, and 42 degrees C) were investigated. Incubation at 37 or 40 degrees C was permissive for HSV-2 inhibition of host DNA synthesis, induction of virus-specific DNA replication, and infectious virus production. The amount of [methyl-3H]thymidine incorporated into viral DNA and the final yield of new infectious virus were significantly reduced at 40 degrees C compared to 37 degrees C. At 42 degrees C, detectable virus-specific DNA synthesis was totally blocked. Maximum stimulation of host cell DNA synthesis at 42 degrees C was measured after a multiplicity of infection of 0.5 to 1.0 PFU/cell. By autoradiography, data indicated that HSV-2 stimulates host cell chromosomal DNA synthesis. Stimulation of thymidine kinase activity with thermostability properties in common with a virus enzyme was detected during the first 24 h of infection at 42 degrees C, after 24 h the enhanced thymidine kinase activity had properties in common with host cell isozymes. The data obtained during this investigation indicated that stimulation of host cell DNA synthesis does not require viral DNA synthesis.


Simian virus 40 DNA replication in nuclear monolayers. Simian virus 40 DNA replication has been studied in nuclear monolayers prepared by treatment of monolayers of BSC-1 monkey kidney cells with Nonidet P-40. These nuclear monolayers incorporated [3H]TTP into two types of viral replicative intermediates that sediment as 25-26S and 22-23S species, respectively, in neutral sucrose gradients. The 22-23S species behaves, in dye buoyant density equilibrium gradients, as a late replicative intermediate. Examination of both species in alkaline sucrose gradients revealed the presence of two types of newly synthesized strands: (i) 4-7S strands and (ii) full-length, or nearly full-length, 10-16S strands. At low TTP concentrations (less than 0.5 muM), the two size classes were found in approximately equal amounts. However, at 10 to 50 muM TTP, the proportion of the longer strands increased, with a corresponding decrease in the relative amount of the 4-7S species. Thus, the joining of small, Okazaki-like fragments to the growing chain appears to require a much higher concentration of TTP than the synthesis of the fragments themselves. Replicating simian virus 40 DNA synthesized in the nuclear monolayers is is associated with "M bands", as previously demonstrated for replicating simian virus 40 DNA in cultured whole cells.


Isolation and structural characterization of monomeric and dimeric forms of replicative intermediates of Kilham rat virus DNA. Two virus-specific species of newly synthesized DNA were isolated from rat fibroblast cell cultures infected with the Kilham rat virus (RV). These two DNA species were purified; their behavior on hydroxyapatite chromatography and their sedimentation coefficients in sucrose gradients were determined. One of the two species corresponds to the linear double-stranded form of the RV DNA, and the other corresponds to the dimeric duplex form. After denaturation, a fraction of both species showed an intramolecular renaturation; these molecules are composed of viral strand covalently linked to complementary strand. Models for the structure of both species are posposed. Both species may be considered as double-strand replicative intermediates of the single-stranded RV DNA.


Origin of polyoma virus-associated endonuclease. Polyoma virus particles purified from infected cells, but not from the culture medium, exhibited an endonuclease activity distinct from the serum contaminant recently described. This endonuclease cleaved form I polyoma DNA once only per molecule, at one of three possible sites corresponding to the known adenosine-ribosylthymine-rich regions of the molecule.


Analysis of intracellular feline leukemia virus proteins. I. Identification of a 60,000-dalton precursor of feline leukemia virus p30. The synthesis and release of feline leukemia virus p30 was studied using a permanently infected feline thymus tumor cell line. Disrupted cells were divided into two subcellular fractions, a cytoplasmic extract (CE) representing cellular material soluble in 0.5% NP-40 and a particulate fraction (PF) insoluble in 0.5% NP-40 but soluble in 0.2% deoxycholate and 0.5% NP-40. Intracellular feline leukemia virus p30 was isolated from infected cells by immune precipitation with antiserum to p30 and subsequent sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the precipitated proteins. Cells labeled for 3 h with [35S]methionine contained equal amounts of p30 in both the CE and the PF. p30 synthesis was estimated to be 0.8% of the total host cell protein synthesis. Immune precipitates from cell pulse labeled for 2.5 min contained a labeled 60,000-dalton polypeptide (Pp60) in the PF and a polypeptide in the CE that comigrated with feline leukemia virus p30 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When cells were chased after a pulse label, there was a rapid loss of Pp60 in the PF and an accumulation of p30 in the CE within 30 min followed by distribution of p30 in both the PF and the CE. Estimation of intracellular and extracellular p30 levels during a 0.5- to 24-h chase period suggested that most of the newly synthesized p30 was incorporated into extracellular virus. Typtic peptide analysis of labeled Pp60 and p30 demonstrated the presence of 13 of 15 p30 peptides within the Pp60 molecule. The tryptic peptide analysis in concert with the pulse-chase labeling data provides strong evidence that Pp60 is a precursor of p30.


Infection of resistant avian cells by subgroup B Rous sarcoma virus. Chicken fibroblasts derived from the H & N flock, which have been characterized as resistant to subgroup B avian oncornaviruses in focus assays, can be infected in suspension shortly after trypsinization by subgroup B sarcoma and leukosis viruses. Once cells are plated, resistance to infection reappears rapidly. C/BE cell suspensions obtained by treatment with EDTA instead of trypsin are not as sensitive to infection. Late interference established by preinfection with subgroup B leukosis viruses is not overcome by trypsinization. In addition to C/BE H & N chicken cells, C/ABE RPRL line 7 cells can also be infected by subgroup B viruses shortly after trypsinization; however, none of the cell types can be made sensitive to subgroup E infection. These results are discussed in relation to current information on the genetic control of resistance to avian oncornaviruses.


Simian virus 40-permissive cell interactions: selection and characterization of spontaneously arising monkey cells that are resistant to simian virus 40 infection. A fraction of permissive cells survive simian virus 40 (SV40) infection. The frequency of such surviving cells depends only upon the concentration of infecting virus, both parental and progeny, to which the cells are exposed during the course of selection. Surviving clones, which can be freed of virus by cloning in the presence of SV40 antiserum, are indistinguishable from parental cells in their growth of characteristics and display no SV40 antigen; thus they are not transformed. Most surviving clones are less than 10% as susceptible as parental cells to SV40 infection; 5 to 10% are less than 1% as susceptible. None of these SV40-resistant clones is absolutely resistant to SV40 infection. Analysis of 16 independently arising resistant clones indicates that they all block SV40 infection at an early stage after adsorption and eclipse but before full uncoating. Viral mutants have been isolated that partially overcome the block to infection in these cells; these host range viruses plaque on resistant lines fivefold more efficiently than wild-type SV40 and have a characteristic plaque morphology. Fluctuation analysis indicates that resistant cells arise spontaneously during the growth of normally susceptible permissive cells. Thus, SV40-resistant cells are selected for, not induced by, SV40 infection.


Herpes simplex virus gene expression in transformed cells. I. Regulation of the viral thymidine kinase gene in transformed L cells by products of superinfecting virus. In this paper we show that the expression of the herpes simplex virus type 1 (HSV-1) gene for thymidine kinase (tk) in HSV-transformed cells is subject to regulation by two viral products synthesized during productive infection of these cells with a tk- mutant of HSV-1. The cell line used in this study is a derivative of tk-deficient mouse L cells that, after exposure to UV-inactivated HSV-1, had acquired the HSV-1 gene for tk (which we term a resident viral gene) and consequently expressed the tk+ phenotype (LVtk+ cells). Productive infection of these cells with HSV-1(tk-) at appropriate multiplicities caused significant enhancement of the viral tk activity. The results of several experiments allow us to conclude that this enhancement was due to increased synthesis of tk specified by the HSV-1 gene resident in the LVtk+ cells and that a specific protein made early after infection with HSV-1(tk-) mediated the enhancement, probably by increasing the production of mRNA from the viral tk gene resident in the LVtk+ cells. Our data also indicate that another HSV-1(tk-) product acted to turn off tk synthesis. The finding that tk activity continued to increase for a longer time after infection of the LVtk+ cells at 2 PFU/cell than after infection at higher multiplicities suggested the synthesis of a product which inhibited tk synthesis and whose concentration reached critical levels earlier at higher multiplicities of infection. Inhibition of DNA synthesis after infection, a treatment that depresses the synthesis of late viral proteins, prolonged the synthesis of tk in LVtk+ cells infected at either 2 or 5 PFU/cell. Infection of the LVtk+ cells with HSV-2(tk-) resulted in only small increases in tk activity, indicating some type specificity in recognition of viral products that can modify the expression of the HSV-1 tk gene resident in these cells.


Mouse strain resistant to N-, B-, and NB-tropic murine leukemia viruses. Mouse strain G was studied for its susceptibility to various strains of murine leukemia and sarcoma viruses. Both N- and NB-tropic Friend leukemia viruses neither induced splenomegaly nor grew efficiently in strain G mice. Using the XC test, cultured embryo cells were found to be resistant, but not absolutely, to all the tested viruses, N-tropic AKR virus, N- and NB-tropic Friend leukemia viruses, NB-tropic Rauscher leukemia virus, B-tropic WN1802B virus, NB-tropic Moloney leukemia and sarcoma viruses, and N-tropic Kirsten sarcoma virus, although the resistance to Moloney leukemia and sarcoma viruses is sometimes not as strong as that for other viruses. Thus, the strain G mice are unique among mouse strains because they show resistance that is not related to the N-B tropism of murine leukemia viruses.


Enzymatic basis for the selective inhibition of varicella-zoster virus by 5-halogenated analogues of deoxycytidine. 5-Bromodeoxycytidine (BrdC) and 5-iododeoxycytidine, at a concentration of 100 mug/ml, effectively inhibit the replication of varicella-zoster (VZ) virus in tissue culture. No toxicity could be demonstrated in uninfected cells under the same conditions. Studies on the enzymatic basis for this selective inhibition were undertaken. Infection of human embryonic lung cell monolayers with VZ virus-infected cells results in the induction of thymidine (dT), deoxycytidine (dC), and BrdC kinase activities (which are increased 10-, 40-, and 60-fold, respectively) and in a 70-fold stimulation in the incorporation of 3H nucleotide (5-bromodeoxyuridylate) derived from BrdC into DNA. The thermal stability of the VZ virus-induced activities differs significantly from the activities induced by herpes simplex virus type 1 and herpes simplex virus type 2 and those present in uninfected human embryonic lung cells. The VZ virus-induced dT, dC, and BrdC kinase are similarly affected by temperature and cofractionate upon Sephadex gel filtration, findings consistent with the hypothesis that these activities are the function of a single enzyme: a pyrimidine deoxyribonucleoside kinase. The molecular weight, calculated on the basis of the elution pattern on Sephadex G-150, is 70,000. Kinetic studies, demonstrating that dT and dC competively inhibit the phosphorylation of BrdC, are consistent with the phosphorylation of these substrates at a common active site. Kinetic parameters include: KidT = 0.6 MUM; KidC = 60 muM; KmBrdC = 8.5 muM. In contrast to its relatively high affinity for the VZ virus-induced kinase, BrdC is a relatively poor substrate for the host kinases. Therefore, the basis for the selective inhibition of VZ virus by 5-halogenated analogues of dC is reflected in the induction of a pyrimidine deoxyribonucleoside kinase with a high affinity for BrdC.


Anatomy of herpes simplex virus DNA. VI. Defective DNA originates from the S component. We previously reported that serial propagation of the Justin strain of herpes simplex virus 1 [HSV-1 (Justin)] results in the generation of defective DNA molecules consisting of tandem repetitions of sequences of limited complexity. In the present study, HSV-1 DNA was cleaved with the restriction endonucleases BglII and EcoRI. The fragments were electrophoretically separated on agarose gels, transferred to nitrocellulose strips, and then hybridized with 32P-labeled HSV-1 (Justin) defective DNA. The data allow us to conclude that DNA sequences contained in the repeat unit of defective DNA originate from the S segment of the wild-type viral DNA molecule.


Two adenovirus type 2-simian virus 40 hybrid populations that contain the entire SV40 genome differ in their ability to induce SV40 transplantation immunity in hamsters but not in mice. Ad2++ HEY and Ad2++ LEY are two adenovirus 2(Ad2)-simian virus 40 (SV40) hybrids distinguished by differences in the efficiency with which they produce SV40 progeny in lytically infected African green monkey kidney cells. These virus populations are composed of nonhybrid Ad2 and hybrid virions, the majority of which contain more than 1 unit of SV40 DNA. The Ad2++ HEY and LEY populations also differ in their ability to induce SV40 transplantation immunity in rodents. Only Ad2++ HEY induces SV40 transplantation immunity in hamsters, whereas both viruses induce significant SV40 transplantation immunity in adult BALB/c mice.


Urologic abdominal masses in infants and children. A series of 167 infants and children presenting with palpable urologic abdominal masses from 1960 to 1973 has been reviewed. The groupings by age at presentation and type of urologic lesion enable us to make certain general statements. Emphasis is placed upon adequate preoperative evaluation in any child with an abdominal mass to avoid unnecessary laparotomy and morbidity.


Cerebrospinal fluid IgG level in herpes simplex encephalitis. Elevated levels of immunoglobulin G (IgG) in the cerebrospinal fluid (CSF) were associated with elevated herpes antibody in the CSF in a case of herpes simplex encephalitis. The case was also characterized by a prolonged course and a difficult virus isolation.


Difference in response to mouse hepatitis virus among susceptible mouse strains. After intraperitoneal inoculation with a high-virulent mouse hepatitis virus (MHV) a significant difference was seen in survival time between DDD and CDF1 (BALB/c X DDD) mice, while 50% lethal doses were not significantly different. With 3 X 10(3) PFU of the virus CDF1 and DDD mice died in 45 and 120 hr, respectively, on the average. This difference of susceptibility between DDD and CDF1 mice was first demonstrable at the age of 1 week and was more pronounced at the age of 4 weeks but showed no dependence of the sex. Virus titers ran 2 to 3 log higher in the liver and blood of CDF1 than in those of DDD mice, while being only 1 log higher in the spleen. At an early stage of infection viral antigen was demonstrable by immunofluorescence in sinusoidal lining cells of the liver more prominently in VDF1 than in DDD mice. Interferon production occurring in parallel with virus growth was significantly higher in CDF1 than in DDD mice. In DDD mice, liver lesions were rather focal with some accumulation of round cells, while they were confluent with poor cellular response in CDF1 mice. Viral growth in cultured peritoneal macrophages from CDF1 mice was 1 log higher than in those from DDD mice. The results suggest that the divergence in response to MHV among susceptible mice greatly depends upon the susceptibility of macrophages and reticuloendothelial cells which constitute primary targets of the virus.


Enhanced growth and plaquing of rabies virus in static chick embryo cell culture. The 7-day egg passage line of HEP Flury strain of rabies virus was inoculated to primary chick embyro (CE) cells prepared in different ways to compared efficiencies of viral growth and plaquing. Special care to minimize cellular damage due to trypsin at the step of monodispersion and sowing a comparatively large number of cells for monolayer preparation were required for rabies plaquing, whereas such cares were not necessary for plaquing of vesicular stomatitis virus. Plaque number and size were increased by incorporation of a high concentration of thymidine into cell growth medium. Various other means to produce a static state of CE cells were tested, and a maximal plaquing efficiency was obtained when dishes receiving a massive number of dispersed cells in MEM plus 1% calf serum were incubated at 37 C for 1 day without any buffering for monolayer preparation and postinfection incubation was done at 32 C in a CO2-incubator. Bottle cultures of CE cells prepared in a similar manner, when infected with HEP Flury virus, yielded a markedly higher titer of virus that CE cells prepared by our previous standard method.


[Comparative characteristics of the lipoproteins of the human vascular wall and blood plasma]. Electrophoresis in the polyacrylamide gel and ultracentrifugation with a multistage density gradient demonstrated virtually the whole of the blood plasma lipoproteids spectrum to be contained in the human aortal wall. An immunological identity of the whole beta- and alpha-lipoproteids and also of their individual apoproteins in the aortal wall and blood plasma was ascertained. Moreover, the ratio of etherified cholesterol/total cholesterol in the vascular and plasmatic lipoproteids proved to be quiet close. In determining the content of the summary pre-beta-and beta-lipoproteids in the aortal wall the quantity of the said lipoproteids in atherosclerotic lesions was found several times as high as in a normal aorta, whereas the lipoproteids concentration in an intact aortal wall correlated with the intensity of atherosclerosis in the same aorta. The data obtained bear evidence to a plasmatic origin of lipoproteids in the vascular wall and to their important role in atherogenesis.


[Use of miskleron in ischemic heart disease with differing types of hyperlipoproteinemias]. The effect of Misclerone (clofibrate) was studied in 127 patients with different types of hyperlipoproteinemia treated with the drug for 1 month and longer. On the basis of the obtained data it was concluded that Misclerone is effective both in patients with isolated disorders in cholesterol metabolism (Type II), and in those with cholesterol and triglycerides metabolism disorders (Types IIb and III), and in those with prevailing triglycerides metabolism disorders (Type IV). The efficacy of the drug in Type II hyperlipoproteinemia is determined by the degree and nature of metabolic disorders of low density lipoproteins.


[Clinical and morphological studies in a case of congenital glaucoma combined with an ocular-dental-digital dysplasia (author's transl)]. Clinical and morphological studies were undertaken to characterize a case of congenital glaucoma combined with an ocular-dental-digital dysplasia in a two year old patient. Light and electromicroscopic investigations of a surgically removed trabecular specimen were carried out. A trabeculotomy had been performed at this exact site 10 months prior to removal. The existence of a canal of Schlemm was demonstrated. The regular trabecular meshwork and trabecular region was replaced by fairly dense scar tissue, containing rudimentary intertrabecular spaces. Proliferating corneal endothelial cells covered the wound area up to the uveal part of the trabecular region. The morphological findings correlated well with the clinical observation of continued raised intraocular pressure despite trabeculotomy. It is concluded that, contrary to the situation in adult onset glaucoma, the strong proliferative tendency of juvenile tissue has a marked influence on the postoperative efficacy of the trabeculotomy.


[Total syndactylism with mesomelic shortening of the arm, radioulnar and metacarpal synostoses and disorganization of the phalanges ("cenani syndactylism") (author's transl)]. An additional case "Cenani syndactylism" in a 7 years-old female child was described. With this autosomal recessive inherited disease, the radius and ulna are severly shortened and generally fused with each other. The metacarpal bones are, for the most part, synostosized. The phalanges are badly disorganized. Usually no fingers can be recognized externally. The alterations in the feet are far less pronounced. Syndactylism and ray reduction are frequent. A similar, but apparently independent, case with radioulnar synstostoses, reduction, dysplasia and fusion of the metacarpal bones but less pronounced malformations of the fingers was also described. Finally, new type of malformation was present for comparison in which an increased number of metacarpi and metatarsi and, to some extent, severely disorganized phalanges with massive syndactylism are present. The etiology of the second and third cases could not be explained.


[Plasma-cAMP in primary and secondary hyperparathyroidism (author's transl)]. 3 patients with operatively proven primary hyperparathyroidism showed normal plasma cAMP concentrations. 5 patients in chronic dialysis treatment independent of the level of parathyroid hormone had increased plasma cAMP concentrations. These observations suggest that there is under these circumstances no regulation of plasma cAMP by the parathyroid hormone.


Physiological responses to stress with reference to the domestic fowl. Environmental stress may effect the outcome of routine investigations as well as experimental procedures. This is illustrated by investigations in the domestic fowl.


Differences in tumor incidence in two substrains of Claude BALB/c (BALB/cfCd) mice, emphasizing renal, mammary, pancreatic, and synovial tumors. Observations were made on the tumor incidences in two substrains of the BALB/CFCd mouse. A total of 900 mice were examined. They comprised two substrains (families), direct descendants of two different females (designated 916 and 917) of the 31st inbred generation. Offspring of sibling matings of the fourth and fifth generation descendants of these two females were killed when moribund or when they had visible or palpable masses. Complete gross and microscopic examinations were conducted. Renal tumors were noted in 48.1% of those necropsied in family 916 and in 24.6% of those in family 917 (p less than .025). Mammary tumors were found in 13.3% of family 917 and in 3.3% of family 916 (p less than .001). Neoplasms of the reticulo-endothelial system (reticulum cell neoplasms, lymphocytic leukemia, other lymphocytic neoplasms) were found in 20.0% of family 917 but only in 11.2% of family 916 (p less than .005). Tumors of the respiratory system (primarily alveolar adenomas and alveolar adenocarcinomas) were found in 10.2% of family 916 and 16.5% of family 917 (p less than .05). Less commonly observed tumors included synoviomas, 7.6% in family 916 versus 2.1% in 917 (p less than .005), and seven pancreatic acinar adenomas, all but one of which were found in family 916. Also recorded were a total of nine myoepitheliomas, 28 hemangioendotheliomas, two interstitial cell tumors of the testis, a single mesothelioma and a single rhabdomyosarcoma of the esophagus. Degenerative lesions consisting of kyphoscoliosis were noted in 3.7% of mice examined in both families combined.


The agouti (Dasyprocta sp) in biomedical research and captivity. Agoutis (Dasyprocta sp) are hystricomorph rodents belonging to the family Dasyproctidae. Utilization of agoutis in a research program of the National Institutes of Health necessitated the development of suitable methods for feeding, housing and collecting blood. The rodents were maintained successfully in conventional dog cages and kennels, and thrived on a diet of primate diet and apples. Physical examinations and periodic blood collections were best performed while agoutis were sedated with ketamine HCl (42-83 mg/kg, intramuscularly). Satisfactory sites for blood sampling were the cephalic, saphenous and jugular veins. Hematologic and serum chemical values established for the agouti were similar to those found in other rodents. Experimentally induced and naturally occurring diseases of agoutis were reviewed.


Biological values of the infant, juvenile, and adult agouti (Dasyprocta sp) with emphasis on microbial flora. Normal values for intestinal flora were determined on four adult, two juvenile, and four infant agoutis (Dasyprocta sp) maintained at our institution. Serologic, hematologic, biochemical, and histologic observations were also made on these same agoutis, and serologic, microbiologic, and endoparasitic tests were made on serum and fecal samples from agoutis maintained at other institutions. Streptococci, lactobacilli and nonenteropathogenic Escherichia coli were isolated from cecal contents of all age groups. Gaffkya tetragena and Bacillus sp were recovered from infant agoutis, Proteus mirabilis from juvenile agoutis, and Proteus mirabilis and Micrococcus sp from adult agoutis. Infant agoutis showed marginal antibody titers to reovirus type 3 and Toolan's H-1 viruses, and Theiler's GDVII antibodies were detected in juvenile agoutis, but no antibody titers to murine viruses were found in the adult agoutis maintained at our institution. Hematologic, biochemical, and histologic data for these agoutis were comparable to those of other hystricomorph rodents. Serologic tests of the agoutis maintained at other institutions disclosed antibodies for GDVII virus in all of 17 animals tested, while marginal titers were found for Sendai virus in four and for reovirus type 3 in eight of the 17 animals. Only low incidences of pathogenic bacteria (Salmonella, Pseudomonas, and Citrobacter) were isolated from the agoutis maintained elsewhere. The endoparasites found in these agoutis were Ascaris, Toxascaris, Strongyloides, Trichuris, and Trichomonas.


Pathogenesis of visna. I. Sequential virologic, serologic, and pathologic studies. A total of 56 Icelandic sheep were infected with visna virus by intracerebral injection of strain 1514 and the course of infection was followed for 12 months. Virus was isolated from more than 90 per cent of the animals, primarily from central nervous system and lymphoid tissues. However, titers of free infectious virus were minimal and virus isolation often required the use of tissue explants. All sheep raised serum-neutralizing and complement-fixing antibodies beginning 1 to 3 months after infection. Differences in neutralization titers against the infecting strain (1514) and a reference strain (796) suggested that antigenic drift might occur during prolonged infection. High cerebrospinal fluid neutralization titers in the spinal fluid indicated local antibody production in the central nervous system. Although the incidence of clinical disease during the 1st year of infection was less that 10 per cent, approximately 80 per cent of the sheep examined had central nervous system histologic lesions of variable severity, which were marked 1 month after infection with little progression during the subsequent year. There was a striking correlation between the severity of central nervous system lesions and the frequency of virus isolations from all tissues. These observations provide detailed base line data on visna infection, suggest some of the mechanisms responsible for the persistence of infection and for the slowness and irregularity of disease occurrence, and form the basis for further experiments on the role of immunologic mechanisms in the pathogenesis of this slow infection.


Data attrition in follow-up studies of alcoholics. Alcoholics who responded to posttreatment follow-up evaluation showed more improvement than those wh did not respond. Thus nonresponders may bias follow-up research tht does not account for missing data. Several procedures are outlined to increase follow-up return.


A telephone follow-up procedure for increasing the effectiveness of a treatment program for alcoholics. The use of outpatient services by alcoholics can be significantly increased by means of a telephone follow-up procedure.


The metabolism of carbohydrate.

The metabolism of carbohydrate. The purpose of this review has been to establish a background in carbohydrate biochemistry as it relates to the human neonate. The emphasis has therefore been placed on the developing human, although many concepts are derived from studies in animal models. Further discussions of the physiology of carbohydrate metabolism are included in the chapters that follow.


Myocardial ischemia.

Myocardial ischemia. Myocardial ischemia results from an imbalance of energy supply and demand. Because of the essentially aerobic nature of myocardial metabolism and the high oxygen extraction from the blood, ischemia is usually equatable with limitation of blood supply. Coronary atherosclerosis is a patchy disorder, and therefore, ischemia usually occurs in segmental fashion throughout the topography of the heart. Ischemia is invariably seen earliest and most intensely in the deep or subendocardial layers of myocardium. Ischemia leads to biochemical disruption, including initiation of glycolysis, which in turn causes electrophysiological and mechanical disturbances. Myocardial ischemia can be induced naturally or experimentally in the human subject in a variety of ways, some of which have been studied in the laboratory.


[Abrikossof's tumor of a toe].

[Abrikossof's tumor of a toe]. A case of Abrikossof tumor in a 33 year old patient presenting a progressive growth of a toe without associated pain or inflamation. There was an important hypertrophy of the affected toe with hyperkeratosis and pseudoelephantiasis. The histologic examination showed a massive infiltration of granular cells in the dermis. The exceptional localisation of the tumor in this case is emphasized.


[Multiple glomangiomas].

[Multiple glomangiomas]. A case of multiple glomangioma in a 25 year old male patient is reported. The lesions started in an eruptive way affecting the upper limbs and trunk. Most of the forty lesions were painless but the larger ones were painful on pressure. Pain was not elicited by cold nor increased venous pressure. The histopathological examination of three lesions revealed a noncapsulated, angiomatous type of glomus tumor with a small number of glomus cells. Sweat glands are often imprisoned by the tumor in the deep dermis. The classification of glomangioma in solitary, multiple regional and disseminated types is discussed and their clinicopathological features are reviewed. The relationships and differential diagnosis with blue-rubber-bleb-naevus and post-traumatic angiomatosis are discussed.


[Prognostic value of histological rare breast cancer (author's transl)]. Among 835 cases of female breast cancer in the years 1951-1971 we found 44 histological rare cancer (comedo-carcinoma 21, carcinoma cribosum 1, gelatinous carcinoma 8, squamous cell carcinoma 3, mixed carcinoma 11). In agreement with the literature we are able to underline the favourable prognosis of the slow growing forms of the neoplasms like gelatinous carcinoma, comedo-carcinoma and carcinoma cribrosum opposite to squamous cell carcinoma and mixed carcinoma.


Glomus jugulare tumors of the middle ear and mastoid: diagnosis and surgical treatment. At the 1976 Southern Sectional Meeting of American Laryngological, Rhinological and Otological Society, Inc., a color movie presenting two cases on the "diagnosis and treatment of glomus jugulare tumors of the middle ear and mastoid" was shown. The purpose of this paper and film is to describe the author's experience of 16 cases of glomus tumors involving the middle ear and mastoid (seven tympanicum and nine jugulare). A review of the literature and findings pertaining specifically to the two cases shown in the film is presented.


Relationships between skeletal maturity and submaximal working capacity in boys 8 to 18 years. Skeletal age (SA) height, weight, PWC130, Vo2 at 130 heart rate (Vo130) and cardiorespiratory adaptation to work at 3 kgm-sec-1 were measured in 237 boys ranging in age from 8 to 18 years. Correlations for the total population for the variables of the study are generally higher with SA than with chronological age (CA), although there is no significant difference between the two sets of correlations. Analysis of linear trend indicate that significant nonlinear components are more frequent in the relationships between the variables and SA than with CA.Multiple regression analyses for each variate as dependent variable indicate that height and body weight generally contribute more to the explained variance, while SA is the next best predictor. CA is the least significant of the four independent variables. The total variance explained between any of the submaximal working capacity related measures acting as dependent variable and CA, height and weight is not significantly affected by the presence or absence of SA. The present study thus indicates that SA is not a significant factor in explaining submaximal working capacity, beyond CA, height and weight, when considered over the 8 to 18 year age span. However, when considering shorter age spans, or when considering the adolescent period (12-16 years), SA correlates better with PWC130 (.320 less than r less than .675) and with Vo130 (.299 less than r less than .772).


Is decreased activity of C-II activated lipoprotein lipase in type III hyperlipoproteinemia (broad-beta-disease) a cause or an effect of increased apolipoprotein E levels? Apolipoprotein E (ApoE; "arginine-rich" polypeptide) strongly inhibited both C-I and C-II activated lipoprotein lipases but not the protamine insensitive triglyceride lipase. Inhibition of lipoprotein lipases by ApoE in contrast to inhibition by C-III was not reversed to any significant extent by either increased concentration of activator or triglyceride in the substrate. Our previous studies have shown that in a type III hyperlipoproteinemia (broad-beta-disease) a post-heparin plasma lipoprotein lipase activated by C-II polypeptide of lipoprotein C is decreased in enzyme activity and exhibits an impaired ability to hydrolyze triglycerides in very low density lipoproteins. Type III patients are characterized by elevated concentrations of ApoE in the serum. The data presented in this report suggest that the decreased C-II activated lipoprotein lipase may be further aggravated by increased ApoE levels. Since this enzyme is involved in the catabolism and removal of lipoproteins, decreased activity of C-II activativated lipoprotein lipase may presumably be responsible for increased ApoE.


Inhibition of actions of glucagon in adipocytes by gastric inhibitory polypeptide. Possible interactions between gastric inhibitory polypeptide (GIP) and glucagon were investigated in rat adipocytes. GIP was nonlipolytic and inhibited lipolysis stimulated by glucagon but not that stimulated by secretin or vasoactive intestinal polypeptide (VIP). GIP competed with 125I-glucagon for binding to adipocyte receptors, and at physiologic concentrations inhibited the stimulation of AMP produced by glucagon. Thus GIP acts as an inhibitor of actions of glucagon on adipocytes and may be a physiologic modulator of effects of glucagon.


Demonstration of the production of human thyroid-stimulating immunoglobulins (HTSI) by Graves' lymphocytes cultured in vitro with phytohaemagglutinin (PHA). The lymphocytes from patients with Graves' disease or from healthy subjects have been cultured in vitro either alone or with phytohemagglutinin (PHA). After six days the culture supernatants have been assayed for their human thyroid-stimulating activity by measuring increases in adenosine 3',5' monophosphate (cyclic AMP) in human thyroid slices with which the supernatants have been incubated. Significant levels of human thyroid stimulator activity were found in the culture in which Graves' lymphocytes were cultured with PHA. This activity has been abolished by precipitation of the IgG from the culture supernatant with goat antihuman IgG serum. In contrast, when Graves' lymphocytes were cultured alone, or when control lymphocytes were cultured either alone or with PHA, there was no overall significant production of human thyroid-stimulating immunoglobulin (HTSI). It is concluded that Graves' lymphocytes can be stimulated by PHA to produce HTSI in vitro. Since PHA is known to stimulate only the T lymphocytes, which do not themselves elaborate immunoglobulins as the B lymphocytes do, the above observations indicate a cooperation between T and B lymphocytes in the production of HTSI, at least in this system.


Chylomicron clearance in normal and hyperlipidemic man. A method has been developed for measurement of fractional clearance rates of chylomicrons in man. The technique employs constant infusion of emulsified fat into the duodenum at a rate of 200 mg/kh/hr. After 5 hr of infusion, concentrations of triglycerides (TG) in the chylomicron fraction become constant for the subsequent 5 hr. Since the input of chylomicron-TG is known, fractional removal rates can be calculated from steady-state plasma levels. In 21 patients with normal TG levels, clearance rates for chylomicrons were extremely rapid (t1/2 for chylomicron-TG equals 4.5 +/- 2.9 (SD) min). In 30 patients with endogenous hypertriglyceridemia, clearance was generally prolonged (t1/2 equals 23 +/- 5.5 min). This delay in chylomicron clearance could have been due either to a defect in removal of all TG-rich lipoproteinarticles; a generalized defect in clearance capacity for plasma TG was apparently ruled out for most patients by the further observation that reduction of endogenous TG by caloric restriction caused chylomicron removal to return to normal. These studies also showed that endogenous-TG is removed much less efficiently than chylomicron-TG, and in some patients, this discrepancy is particularly marked.


The distribution of plasma lipoproteins in middle-aged male runners. Stimulated by increasing evidence of an inverse relationship between plasma high-density lipoprotein cholesterol level and frequency of coronary heart disease, we determined concentrations of fasting plasma cholesterol, triglyceride, and lipoproteins in 41 very active men (running greater than 15 miles/wk for the previous year) 35-59 years of age (mean age, 47) and in a comparison group of men of similar age, randomly selected from three northern California communities. The runners had significantly (p less than 0.05) decreased mean plasma triglyceride (70 versus 146 mg/100 ml), total plasma cholesterol (200 versus 210 mg/100 ml), and low-density lipoprotein (LDL) cholesterol (125 versus 139 mg/100 ml) concentrations, and a higher mean level of high-density lipoprotein (HDL) cholesterol (64 versus 43 mg/100 ml) than the comparison group (n equals 147 for HDL and LDL; n equals 743 for total cholesterol and triglycerides). These very active men exhibited a plasma lipoprotein profile resembling that of younger women rather tan of sedentary, middle-aged men. This characteristic, and apparently advantageous, pattern could be only partially accounted for by differences in adiposity between runners and control subjects.


Changes in plasma triglyceride metabolism during withdrawal of bile. The relationship between bile acid turnover and plasma triglyceride metabolism was examined in a group of seven moderately overweight and hypertriglyceridemic men. Bile acid synthesis was stimulated by the continuous aspiration of duodenal contents so that 150% of the bile acid pool was removed in 18 hr. In comparison with the average concentration of very low density lipoprotein triglyceride (VLDL-TG) during a preceding 8-10 hr control period, bile withdrawal led to an increase in VLDL-TG of 34%, 43%, 32%, 14%, 26%, 14%, and 8% (mean +24%) in the seven men. In control studies performed on another day the mean change over a similar period of time, but without bile withdrawal, was +1% (p less than 0.01). The kinetics of VLDL triglyceride were studied in some of the subjects by analyzing the triglyceride specific radioactivity curves following injections of radiolabelled glycerol. In control studies of 26 hr duration the curves were resolved into two apparent exponential functions, the second part of the curve appearing between 15 and 18 hr after pulse-labeling. During bile withdrawal, the second apparent exponential developed sooner, within 4 hr of beginning the aspiration and coincided with the rise in triglyceride concentration. The fractional rate of the second exponential was at least as fast during bile withdrawal as during the control period despite the increase in VLDL-TG pool size, suggesting that the turnover of triglyceride was higher at a time when bile acid production was being stimulated.


Pediatric familial type III hyperlipoproteinemia. Familial type III hyperlipoproteinemia is almost always first diagnosed in adults; the two 16-yr-old probands with type III in this report represent only the fifth and sixth children reported with the disorder. S.E., a 16-yr-old female, and C.H., a 21-yr-old male, (with palmar xanthomas at age 16) had beta-migrating abnormal very low density lipoproteins (beta-VLDL), density less than 1.006. S.E.'s brother (age 21) and mother (age 57) had type III; her grandmother had beta-VLDL and elevated triglycerides, and a maternal uncle had type IV hyperlipoproteinemia. C.H.'s father had type III; a sister and paternal aunt had type IV hyperlipoproteinemia. It is important to consider the diagnosis of type III hyperlipoproteinemia in the pediatric age group, particularly in children from families with type III hyperlipoproteinemia.


Studies on the role of cAMP-dependent protein kinase in the actions of glucagon and catecholamines on liver glycogen metabolism. Glucagon causes a rapid activation of cAMP-dependent protein kinase in rat liver parenchymal cells which correlates well with the accumulation of cAMP. Full activation of phosphorylase or inactivation of glycogen synthase is achieved with half-maximal or less activation of protein kinase. Epinephrine stimulates glycogen breakdown in these cells mainly by mechanisms involving alpha-adrenergic receptors and not beta-receptors. Activition of alpha-receptors results in rapid activation of phosphorylase and inactivation of glycogen synthase without accumulation of cAMP or activation of cAMP-dependent protein kinase. Activation of beta-receptors causes a transient rise in cAMP and a short-lived activation of protein kinase with correspondingly little stimulation of glycogenolysis.


Effect of actinomycin D on plaque formation by mengovirus. Treatment of L cell monolayers with actinomycin D resulted in an increase in both the size and number of mengovirus plaques when compared to untreated cells. Addition of the drug to the agar overlay was as effective as pretreatment provided the addition was immediately after virus adsorption. The increased plaque number observed was apparently not due to rescue of defective particles or inhibition of interferon production. The data indicate that more mengovirus particles can successfully complete the infectious cycle in L cells in the presence of actinomycin. The data also suggest that indigenous C-type particles in L cells may interfere with picornavirus replication.


Fatal cadmium-fume pneumonitis.

Fatal cadmium-fume pneumonitis. A 38-year-old man developed acute respiratory distress several hours after welding cadmium-plated drums without taking precautions against the inhalation of fumes. His respiratory distress worsened over the ensuing three and a half days, and he died. Histological examination of the lungs showed changes of acute cadmium-fume pneumonitis, and chemical analysis of lungs and liver provided confirmatory evidence of considerable absorption of cadmium, of the order previously recorded as causing death. This case is reported so as to renew awareness of this condition, as the cause of the illness was not diagnosed during life.


[Localization of energy generators in methane oxidizing bacteria]. Cytochromes a, b and c, and their quantitative distribution in the cells, were studied by means of differential spectra in obligate methane oxidizing bacteria, Methylosinus trichosporium with the serine pathway of methane carbon assimilation and Methylomonas agile with the ribulose phosphate pathway of methane carbon assimilation, and different types of topography of intracytoplasmic membranes. The membranes are involved in processes of coupled respiration which was confirmed by cytochemical reactions employed for studying the terminal step of the electron transport chain and Mg2+-stimulated ATPase.


[The effect of etiroxate on serum lipids after myocardial infarction or in angina pectoris]. The lipid-lowering effect of etiroxate was studied in a double-blind cross-over trial comprising 183 patients with hyperlipoproteinaemia, most of whom had suffered myocardial infarction or exhibited confirmed coronary insufficiency. Etiroxate (40 mg daily) and placebo were administered for a period of 12 weeks each. In approximately 60% of the cases with Type IIa/IIb hyperlipoproteinaemia the cholesterol level fell by more than 10%. As regards the frequency of heart complaints, the consumption of nitroglycerine capsules, and heart rate, there were no statistically significant differences between the placebo and etiroxate phases. Thus etiroxate did not have any unfavourable cardiac side-effects in the patients of this study. No negative effects on hepatic and renal function or the peripheral blood count were observed.


[Lymphoblastomic sarcoma of the Burkitt type (the so-called "white Burkitt tumor") (author's transl)]. We report about a lymphoblastomic sarcoma of the Burkitt type with a seventeen year old young man. The Burkitt lymphoma occupies a special position among the malignant tumours and occurs rarely in temperate zones. From the clinical and histological point of view, we found the typical image of a unilocular Burkitt tumor. It was not possible to verify an elevated titre of antibodies against the Epstein-Barr virus.


[Group Y meningococci as respiratory pathogens (author's transl)]. Serogroup Y meningococci are more frequently respiratory pathogens than one might suppose from their prevalence in meningitis and septicemia. A case of a peracute fatal septicemia caused by group Y meningococci is reported here in which post mortem hemorrhagic lesions of the pharyngeal, laryngeal and tracheal mucosae as well as a beginning pneumonia and pleurisy have been found. The association of meningococcal and respiratory viral infections is discussed.


SV40 DNA sequences as an example of the structure of genes functioning in animal cell nuclei. Recent studies of the structure of messenger RNA have demonstrated the existence of untranslated sequences of the 3' and 5' end of the messages. In addition analysis of transcription in vitro has indicated that the nucleotide sequence U6 purine may be part of a transcription termination signal in prokaryotes. Recently it has been possible to determine the sequence of extensive portions of the DNA of SV40 virus. This article reviews the analogies between certain of these sequences and sequences available from prokaryotic messengers and DNAs. Unusual structures, including blocks of AT-rich and GC-rich segment sections and symmetric regions in the DNA near the origin of DNA replication, have been demonstrated and the distribution of stretches of 6 or more deoxyadenylic acids in the DNA of SV40 is consistent with some rho for these sequences in animal cells, either as terminators of transcription or as sites where degradation of transcripts is initiated or sites related to the selective rejection or degradation of transcipts.


Poly ADP-ribosylation of DNA-dependent RNA polymerase I from quail oviduct. Dependence on progesterone stimulation. Progesterone causes in goblet cells of oviducts of estrogen hormone-stimulated immature quails selectively gene activation without affecting DNA synthesis. This biological model has been used to study the influence of poly ADP-ribosylation during the processes of DNA transcription. Administration of progesterone in vivo causes an increase of the activity of RNA polymerase I and II in isolated nuclei. This increase is accompanied by a marked decrease of the specific activity of poly (ADP-Rib) polymerase. After in vitro ADP-ribosylation of nuclear proteins the template capacity of chromatin for ""exogenous'' RNA synthesis (with E. coli DNA-dependent RNA polymerases) as well as for ""endogenous'' RNA synthesis with DNA dependent RNA polymerases II is not affected, whereas the data presented seem to indicate that the capacity for RNA synthesis mediated by ""endogenous'' DNA-dependent RNA polymerase I might be inhibited after ADP-ribosylation. Evidence is presented to show that a considerable amount of poly (ADP-Rib), synthesized by poly (ADP-Rib) polymerase in isolated nuclei, is linked with RNA polymerase I. The rate of synthesis of poly (ADP-Rib) is dependent on the incubation temperature (optimum at 25 degrees C) and it can be inhibited by the specific inhibitors of poly (ADP-Rib) polymerase nicotineamide, thymidine and formycin B. Poly (ADP-Rib) is probably associated with RNA polymerase I through a covalent linkage. ADP-ribosylated RNA polymerase I has been purified 550 fold with respect to the nuclear extract corresponding to a 4,000 fold purification from the whole cell homogenate. The ratio between poly (ADP-Rib), formed during preincubation of nuclei with NAD, and RNA polymerase I remains almost constant during the purification procedures. The extent of ADP-ribosylation of RNA polymerase I decreases during gene expression. Thus we conclude that poly ADP-ribosylation of this enzyme is one of the regulatory mechanisms by which specificity of DNA transcription is achieved.


Integration of biochemical functions of different cells of rat gastric mucosa for hydrochloric acid secretion. The regulation patterns of gastric acid secretion in rats were investigated. Pentagastrin and histamine stimulate gastric acid secretion, but the inhibitors of DNA-dependent synthesis of RNA and of proteins prevent only the pentagastrin action. It has been found that pentagastrin induces histidine decarboxylase in gastric mucosa, ensuring local accumulation of histamine. The latter activates adenylate cyclase and results in 3',5'-AMP accumulation in gastric tissues. The administration of pentagastrin, histamine or 3',5'-AMP enhances the activity of gastric carbonic anhydrase, the enzyme which takes part in HCl formation. The data suggest that these three compounds act sequentially (pentagastrin leads to histamine leads to3',5'-AMP) and the effect of the last one could be mediated through 3',5'-AMP dependent protein kinase. The experiments in vitro demonstrated that gastric carbonic anhydrase can be separated into two isoenzymes and thephosphorylation of one of them by the 3',5'-AMP dependent protein kinase sharply increases its activity. The findings raise the possibility that histamine and 3',5'-AMP, mediating gastrin action, form together with enzymes (histidine decarboxylase, adenylate cyclase, protein kinase, carbonic anhydrase) a caascade of amplifiers. Autoradiographic studies have shown that [3H]-pentagastrin is not bound by oxyntic cells but adheres preferentially to histamine-producing alpha-like endocrine cells and to the chief cells, while 3H-histamine adheres preferentially to oxyntic and to chief cells. Electron microscopy indicates that only pentagastrin (but not histamine) initiates in alpha-like endocrine cells ultrastructural changes characteristic for induction. Pentagastrin, histamine and 3',5'-AMP administration produces in oxyntic cells ultrastructural changes typical for the secretion processes. These results lead to assumption that pentagastrin (gastrin) induces histidine decarboxylase in alpha-like endocrine cells of gastric glands. Histamine which is secreted enhances adenylate cyclase activity in the neighbouring oxyntic cells where 3',5'-AMP dependent protein kinase activates carbonic anhydrase by means of phosphorylation. These different cells form, probably, a multicellular functional unit for gastric acid secretion.


[Therapy of Childhood Asthma with Beclomethasone Dipropionate (author's transl)]. In addition to the usual mangement of asthma the introduction of Beclomethasone-dipropionate aerosol can be considered as a progress in the long-term treatment of asthma, because in most cases oral and parenteral application of corticosteroids is no longer necessary. According to the results of many authors the wellknown side effects of steriod therapy have so far not been observed. We report our experiences with Beclomethasone in the treatment of 19 chronically asthmatic children.


[Screening of newborns for inborn errors of galactose metabolism. Methods and results]. Three inborn errors of galactose metabolism as known today. Only two of them cause illness: the deficiencies of galactokinase and of galactose-l-phosphate uridyltransferase. Both can be detected through mass screening of newborns and are amenable to a simple and successful dietary treatment. Mass screening of newborns for disorders of galactose metabolism can be performed efficiently and inexpensively if it is incorporated in the general newborn screening program.


[Preternatural anus and its care (author's transl)]. The most careful surgical technique, the guiding principles of which are outlined, is a prerequisite for the subsequent possibilities for correct care of preternatural anus and for preventing otherwise unavoidable complications. Four different programs are available to us for care; truss pad, adhesive bag, irrigation therapy and the Erlanger magnetic closure. For prophylaxis of frequent hernias and prolapses, wearing a made-to-measure body binder is indispensable. Skin lesions, formerly feared concomitant symptoms, can now be prevented or eliminated as a result of the development of non-irritant adhesives, combinations with sterculia rings or adhesive plasters and differentiated skin care products. A more comprehensive organization of those with preternatural anus within the framework of the German Ilco and the establishment of preternatural anus clinics and therapists would be desirable.


Interaction between serum factors and T lymphocytes in Hodgkin's disease. Use as a diagnostic test. We studied the mechanism of defective binding of sheep erythrocytes to the surface of peripheral blood T lymphocytes (E-rosette-formation) in Hodgkin's disease. The decreased percentage of E-rosette-forming cells (range, 21 to 77 per cent) in 25 patients with Hodgkin's disease was reversed and returned to normal range (52 to 78 per cent) by prior incubation of the T lymphocytes in tissue-culture medium with 20 per cent fetal-calf serum for 18 to 24 hours. The percentage of E-rosette-forming cells was suppressed by additional incubation with serum from patients with Hodgkin's disease (range, 20 to 61 per cent) but not with serum from patients with other neoplasms or from normal subjects (range, 52 to 74 per cent). Only target T lymphocytes from patients with Hodgkin's disease were suppressed by Hodgkin-disease serum. The findings suggest that there is a specific interaction between serum factors and the surface of peripheral blood T lymphocytes in Hodgkin's disease.


Susceptibility of chickens to avian encephalomyelitis virus. IV. Behavior of the virus in laying hens. Some laying hens 6 months of age were inoculated subcutaneously or orally with a chick embryo--adapted strain of avian encephalomyelitis virus and examined for propagation of the virus in the body. When inoculated subcutaneously, the virus appeared in liver, spleen, ovarian follicle, and muscle at the site of inoculation 1 day, in kidney and lumbar part of the spinal cord 3 days, in the pancreas 5 days, in heart, duodenum, and cervical part of the spinal cord 7 days, and in the brain 11 days after inoculation. After its appearance, it increased gradually in amount in liver, spleen, pancreas, muscle at the site of inoculation, and cervical and lumbar parts of the spinal cord, but remained at a low level in any other organ. When examined 14 days after inoculation and later, it was distributed mainly in the central nervous system. It was detected from 12 of 16 organs examined. The highest virus level in each organ was 10(2.6)/0.1 g in pancreas and lumbar part of the spinal cord, which were followed by muscle at the site of inoculation (10(2.0)/0.1 g), spleen (10(1.8)/0.1 g), cervical part of the spinal cord, heart, and liver in the order listed. When inoculated orally, the virus was found sporadically in spleen, pancreas, kidney, cecum, ovarian follicle, and lumbar part of the spinal cord. The virus level was low in these organs, of which pancreas, kidney, and lumbar part of the spinal cord showed the highest virus level, or 10(1.3)/0.1 g.


Susceptibility of chickens to avian encephalomyelitis virus. V. Behavior of a field strain in laying hens. Some 12-month-old laying hens were inoculated orally or subcutaneously with 10(4.7) EID50 of a field strain of avian encephalomyelitis virus. They were examined for propagation of the virus in the body at regular intervals of time. When two hens were sacrificed daily in the group of oral inoculation, the virus was found in liver, pancreas, and esophagus in both hens 1 day, in brain, lumbar part of the spinal cord, heart, spleen, pharynx, larynx, glandular stomach, muscle, and blood in one of the two hens 1 day, and in various parts of the body 3 approximately 9 days after inoculation. After that, the virus was detected almost continually from the central nervous system and abdominal parenchymatous organs in nearly all the hens examined up to the end of the observation period, or 21 days after inoculation. Virus detection from the digestive tract and ovarian follicle, however, decreased in frequency and virus titer was reduced remarkably with the lapse of time after inoculation. When the largest amount of virus was determined in each organ, it was the largest, or 10(6.5) EID50/0.1 g, in the liver and about 10(5.0) EID50/0.1 g in spleen, pancreas, kidney, and ovarian follicle. There was little difference in virus propagation and its course between the group of subcutaneous inoculation and that of oral administration.


Experimental studies on vertical infection of mice with Japanese encephalitis virus. I. Effect of mouse strain on placental and fetal infection. An experiment was carried out with mice to study the effect of the mouse strain on the establishment of experimental vertical infection with Japanese encephalitis virus. Mice of various inbred strains were inoculated with a field strain of virus on the 7th day of pregnancy. An attempt was made on them to recover the virus from the placenta and fetuses. As a result, there were differences in the rate of placental and fetal infection among the mouse strains used. It was clarified that vertical infection could be established experimentally in some selected mouse strains. There was not always a parallel relationship between the rate of placental and fetal infection and the severity of viremia or visceral infection in the dam.


Cell surface antigens detected in cell lines established from lymphomatous Papio hamadryas and Macaca arctoides monkeys. Cell surface antigens associated with bone marrow cell cultures from leukemic monkeys of species Papio hamadryas and Macacus arctoides were visualized by means of an indirect immunofluorescence method with sera from leukemic baboons. The same immune serum gave two types of immunofluorescence, depending on the origin of the target cells. Fluorescence of the ring-reaction type was seen with Papio hamadryas bone marrow cell cultures growing in suspension and containing the baboon herpesvirus, whereas the patchy type of fluorescence was noted with monolayer bone marrow cell cultures from Macaca arctoides origin, containing type-C oncornavirus but not the herpesvirus particles. Absorption experiments showed that antibodies responsible for the patchy type of immunofluorescence could be absorbed with a disintegrated type-C baboon oncornavirus, and not with baboon lymphoblastoid cell lines containing the herpesvirus nor with human lymphoblastoid cell lines containing the Epstein-Barr virus.


Culture of human cells obtained with DNA from chick Rous sarcoma. An infectious process was reproduced in the culture of chick embryo cells by means of DNA isolated from Rous chick sarcoma tissue (Carr-Zilber strain). This DNA preparation displays biological activity also in the culture of human embryo diploid cells (HEDC) which is manifested in: 1. discontinuous synthesis of avian oncovirus group-specific antigen; 2. enhancement of proliferative activity and morphological transformation of human cells; 3. continuous presence of virus-specific sequences as revealed by DNA/RNA hybridization. Producing complete oncornavirus by means of DNA isolated from Rous chick sarcoma in HEDC was unsuccessful. DNA preparation from gs negative chick embryo cells shows no infectious activity in HEDC culture.


Humoral antibodies to the capsid antigen of Epstein-Barr virus in Hodgkin's disease. By the method of indirect immunofluorescence it has been shown in P3HR-I cells that sera from patients with Hodgkin's disease contain high titers of humoral antibodies to the capsid antigen of Epstein-Barr virus (EBV). Higher titers of antibodies of EBV were found in histological variants of Hodgkin's disease with an unfavorable course. The variant of lymphocyte depletion is accomplished by higher titers of the virus and poorer prognosis than the nodular-sclerotic variant having a course with lower titers of antibodies and better prognosis. At the same time, the level of antibodies does not depend on the results of the therapy applied. In the sera of patients with reticulosarcoma or lymphosarcoma no increase in the level of antibodies to EBV has been discovered in comparison with a group of healthy donors; in acute leukemia a certain tendency to decrease in the level of antibodies to this virus can be observed.


A study on the relation between the Epstein-Barr virus and some forms of malignant tumors in children. The presence of antibodies to the virus capsid antigen of the Epstein-Barr virus was established in the sera of children from different forms of neoplasms with the aid of the indirect method of immunofluorescence according to Henle. 69 sera were studied from children with Wilm's tumor, teratoblastoma, reticulosarcoma, neuroblastoma, sarcoma and also from children with benignant tumors. As control served sera from healthy children of corresponding age. As test cells synthesizing the virus capsid antigen the authors utilized a suspension culture of P3HR-I cells, being one of the clones of Burkitt's lymphoma. The performed investigations have shown that in no one group of children with tumor could there be discovered an increase in the content of antibodies to the Epstein-Barr virus in comparison to controls. It has also been revealed that the spread of the Epstein-Barr virus in different groups of patients and healthy children fluctuated between 83 and 100%.


[Oily contrast medium in liquor space following myelography. A clinical-radiological study on extent, velocity and ways of elimination of monoiodine stearate]. In 32 patients neurological and radiological studies were done after spinal myelography using 5 ml of monoidostearate. The route and speed of elimination of the contrast medium remaining intrathecally have been observed over 2 years, establishing correlations with the way and conditions of administration. In the case of active lumbar drainage from the puncture channel after lumbar instillation of contrast medium or after lumbar puncture, a quick and sometimes massive extrathecal and extravertebral leakage, reaching 1/3 of the applied contrast medium, has been observed in 80%-90% of the cases during the first days and weeks. During the first year, the mean contrast medium elimination was 50%. In the case of a single cisternal instillation of contrast medium, no leakage was found, the elimination was 0-8-1-0 ml/year. The importance of "lumbar puncture drainage" becomes evident, as well as its activity which lasts up to 19 days. The possible routes of leakage (migration) of the contrast medium are discussed.


Proposal of a new model with dopaminergic-cholinergic interactions for neuropharmacological investigations. The motor system of Dugesia gonocephala shows a striking similarity with the extrapyramidal system of high vertebrates and of man with the evidence of correlations between dopaminergic and cholinergic neurons. The utilization of this model seems to be useful in testing drugs which presumably act on dopaminergic or cholinergic transmission. In this model, the quantification of animal behaviour seems considerably easier when compared with the difficulties met in other animal models commonly employed. Besides, it might be anticipated that this model, if correctly used, can display interesting perspectives also in neuroendocrinological investigations.


Two cases of glomus tumour treated by unusual embolization. Two cases of glomus tumour treated by the catheterization technique are reported. In the first case, after it was found impossible to catheterize the external carotid artery by the femoral route, the external carotid was punctured directly, became spasmodic and the catheter could not be introduced, so the inner wall of the external carotid was eroded by manipulating the tip of the metallic guide. This resulted in complete occlusion of the artery 1 cm above the bifurcation of the common carotid. The second case was first treated by embolization of the ascending pharyngeal artery, but with only transitory improvement, so, after 2 months, additional embolization was performed via the vertebral artery, which was followed by immediate and lasting improvement.


Glioblastoma multiforme and the meningeal syndrome. Brain tumors rarely may produce acute steile meningitis (the meningeal syndrome) resulting from the spillage of blood, lipid products of tumor necrosis, or malignant cells into the cerebrospinal fluid (CSF). The frequency of the associated meningeal syndrome is a function of tumor type and of the tproximity of tumor necrosis to the ventricles. The meningeal syndromes of lipid-induced chemical inflammation are seen most commonly with epidermoids, craniopharyngiomas, and infarcted pituitary adenomas. I report a patient with the rare association of the meningeal syndrome with glioblastoma multiforme. The lipid irritants of glioblastomas and craniopharyngiomas are similar chemically and can be detected in the CSF. The anti-inflammatory and immunosuppressant properties of steroids provide a rational basis for their efficacy in treatment of the syndrome.


Herpesvirus hominis type II infections in asymptomatic pregnant women. This study was designed to determine the incidence of previous and current herpesvirus hominis Type II infections in asymptomatic pregnant women. Sera were obtained from 985 patients for detection of Type I and II herpesvirus hominis antibodies. In addition 770 cervical and 211 amniotic fluid cultures for herpesvirus hominis Type II were performed. Identification of previous herpesvirus hominis infection was determined in 352 patients (35.7%) by calculation of a Type II/I antibody index of 85 or greater. Type II herpesvirus hominis was isolated from five cervical cultures for a 0.65% antepartum incidence of infection. None of the amniotic fluid cultures demonstrated virus. The significance of these findings is discussed.


Malignant mixed germ cell tumors of the ovary. A clinical and pathologic analysis of 30 cases. The clinical and pathologic features of 30 ovarian mixed germ cell tumors (neoplasms containing combinations of malignant germ cell elements) were studied to determine their behavior and to compare them with pure forms of malignant germ cell tumors. Dysgerminoma was the most common constituent, found in 24 (80%), followed by endodermal sinus tumor in 21 (70%), teratoma in 16 (53%), choriocarcinoma in 6 (20%), and embryonal carcinoma in 5 (16%). The actuarial survival for the entire group was 46%, and for patients with Stage I tumors it was 50%. The most important factors in predicting the prognosis for patients with Stage I disease was the size and the histologic composition of the neoplasm. If more than one-third of a Stage I neoplasm was composed of endodermal sinus tumor, choriocarcinoma, or Grade 3 teratoma, the prognosis was poor, whereas if the tumor contained less than one-third of these components or contained combinations of dysgerminoma, embryonal carcinoma, or Grade 1 or 2 teratoma, the prognosis was excellent. All patients whose neoplasm was less than 10 cm in maximum diameter survived, regardless of the composition of the tumor. Positive pregnancy tests in nonpregnant patients reflected the presence of either frank choriocarcinoma or scattered syncytiotrophoblastic giant cells. The latter did not appear to alter the prognosis. The finding of elevated serum levels of human chorionic gonadotropin (hCG) in 38% of the nonpregnant patients suggested that serial serum assays for hCG might be useful in staging and monitoring the response to treatment in these patients.


Prostaglandin regulation of adenylate kinases purifed from liver, skeletal muscle, and hepatoma. Adenylate kinases, purified from adult rat liver and skeletal muscle and from a fast growing hepatoma, were examined in the presence of a series of prostaglandins. Prostaglandins A1, A2, E1, E2 and F2alpha stimulated liver adenylate kinase 27% to 149%. All of these protaglandins stimulated the skeletal muscle enzyme from 47% to 82%. While prostaglandins A2 and E2 stimulated, prostaglandins A1 and F2alpha inhibited, and prostaglandin E1 was without effect on the hepatoma adenylate kinase.


Cell coat in tumor cells -- effects of trypsin and EDTA: a biochemical and morphological study. The effects of trypsin and EDTA on the cell coat of ascites tumor cells were studied by means of biochemical and electron microscopy techniques. EDTA seems to release, by chelation of the Ca2+-bridges, an outer layer of glycoproteins of presumably exogenous origin. On the contrary, trypsin produces a deeper enzymic cleavage which appears to affect the structural integrity of the bilayered cell membrane. The significance of the cell leakage in tumor cells and the effect of EDTA on the modification of this leakage by change of cell membrane permeability are discussed.


Problems in polydactyly of the foot.

Problems in polydactyly of the foot. 1. Polydactyly should be treated early. 2. Adequate bone and soft tissue should be removed. 3. One arm of the Y should be removed surgically in the incompletely duplicated Y metatarsal. 4. The incompletely duplicated Y metatarsal shaved to form a single shaft may at first be bowed, but will remodel to form normal or nearly normal bone. 5. The wide metatarsal head associated with a duplicated phalanx should be narrowed surgically to avoid the development of a painful bunion. 6. Short block-like metatarsals remain abnormal. 7. Polydactyly associated with a short first metatarsal and congenital hallux varus carriers a poorer treatment prognosis and may require continuing treatment. 8. Postoperative casting and taping should be utilized to prevent angular deformities and encourage normal forefoot contour.