Ultrastructural evidence for neural crest origin of the malanotic neuroectodermal tumor of infancy. A case of malanotic neuroectodermal tumor of infancy, which occurred in the anterior maxilla of a 3-month-old girl, was studied by  electron microscopy. Polymorphic cellular elements were revealed to be present: large melanin-producing cells with some of the characteristics of a neuroepithelium, small non-pigmented cells resembling immature neuroblasts, and many intermediate forms resembling differentiating neuroblasts. All cell types clearly demonstrated ultrastructural features of neurogenic cells, e.g. neurite-like cytoplasmic processes, fine filaments and microtubules indistinguishable from those seen in normal neurites, synaptic-like structures, and neurosecretory-like vesicles. Based on these fine structures, the lesion was presumed to represent a hamartoma of primitive neuroectodermal cells with a tendency toward the differentiation into melanin-producing cells. The neural crest origin of this tumor was convincingly supported by ultrastructural analysis.


The relation of cyclic AMP levels to phagocytosis and enzyme release in acute inflammation in vivo. The effect of altering endogenous leucocyte cyclic AMP levels on phagocytosis and lysosomal enzyme release was studied in vivo. Acute pleural exudates were produced in rats using either calcium pyrophosphate dihydrate crystals or rat serum. Despite marked increases in leucocyte cyclic AMP concentration produced by injection of dibutyryl cyclic AMP and theophylline, there was no reduction in crystal phagocytosis or in enzyme discharge.


Pathology of ovine adenovirus type 4 infection in SPF lambs: pulmonary and hepatic lesions. Seven lambs were challenged with an aerosol of OA4 virus. Four developed mild pulmonary oedema and a sparse accumulation of mononuclear cells around blood vessels similar to results reported previously. In addition, three had focal hepatic necrosis and one of these three had a non-suppurative occlusive cholangitis and portal tract lymphangitis and thrombosis. Basophilic intranuclear IB's were identified in many necrotic hepatocytes, in lymphatic endothelial cells involved in thrombi but in only one bile duct epithelial cell. Ultrastructurally, virions with the characteristics of adenovirus were seen in hepatocytes. Although only one IB was seen in the affected biliary system there was no evidence that it was caused by an agent other than OA4. Virus was considered to infect the respiratory system directly, be swallowed to infect the alimentary system and carried, possibly within cells, via a haematogeneous and/or lymphogenous route to the liver. The effect of naturally acquired virus on sheep is unknown but it is possible that it may predispose the host to more serious infections.


Electrical excitability of oat cell carcinoma. Cultured tumour cells from two oat cell carcinomas of the lung were demonstrated to generate all-or-nothing, short-duration action potentials similar to those previously reported in neurones, in bronchial carcinoid cells, and in a number of endocrine cells of proven or proposed neural ancestry. This observation provides new evidence to establish oat cell carcinomas and carcinoids as closely related tumours distinct from other pulmonary neoplasms, and raises the possibility of treating these tumours by manipulation of hormonal agents.


Schistosoma mansoni: ultrastructural studies on the esophageal secretory granules. The posterior portion of the esophageal gland of Schistosoma mansoni produces a granule that is highly structured internally. Each granule consists of arrays of membrane-bound tubules enclosed by a membrane. Cytochemical tests indicate that the granules are not reactive for cytochrome c-oxidase but du react for macromolecular carbohydrates. It is believed that the granules are synthesized in the Golgi complex and are secreted at the base of the luminal amplifications of the esophagus. Colchicine treatment results in an accumulation of granules in the cyton region. Their physiological function is still undetermined, but it is hypothesized that they are involved with early stages of digestion of host red blood cells.


Facilitatory and inhibitory effects of gamma-aminobutyric acid on ganglionic transmission in the sympathetic cardiac nerves of the dog. Effects of gamma-aminobutyric acid (GABA) on ganglionic transmission in the peripheral course of the sympathetic cardiac nerves were investigated in vagotomized and cardiac decentralized open-chest dogs. GABA (1-300 microgram/kg) was given i.v. during electrical stimulation of pre-or postganglionic fibers which induced a sustained acceleration of sinus rate. GABA in small doses of 1 and 3 microgram/kg augmented the sinus acceleration during electrical stimulation of the ansa subclavia which largely consists of preganglionic fibers, but depressed the sinus acceleration in large doses over 30 microgram/kg. With a dose of a 10 microgram/kg, its effect was dual and varied from preparation to preparation. On the other hand, GABA did not modify basal heart rate or the increase in heart rate in response to stimulation of the stellate cardiac nerve postganglionic fibers. These results clearly demonstrated dual effects of GABA on ganglionic transmission, i.e., facilitation in small doses and depression in large doses. The depression caused by large doses of GABA was markedly reduced by picrotoxin, 1 mg/kg, while the facilitation remained unaffected. Treatment with atropine, 1 mg/kg, and phenozy-benzamine, 1 mg/kg, failed to influence the effects of GABA. The possible mechanisms for these effects of GABA are discussed.


Antiproliferative activity of anti-inflammatory drugs in two mammalian cell culture lines. The nonsteroid anti-inflammatory drugs inhibited cell proliferation when added to rat hepatoma and human fibroblast cultures. The inhibition was reversible; normal growth resumed when the cultures were washed free of drug. Protein and nucleic acid synthesis, as measured by isotope incorporation was also reduced, although this reduction was probably a reflection of the decrease in cell numbers. An exception was that, in low concentration, the salicylate drugs, salicylamide, salicylic acid and aspirin, stimulated protein and nucleic acid synthesis, but in high concentrations (greater than 1 mM) they inhibited culture growth as well as protein and nucleic acid synthesis. Pharmacologically inactive derivatives, such as m-hydroxybenzoic acid and gentisic acid, were not inhibitory in concentrations up to 5 mM. The order of potency in inhibiting culture growth, meclofenamate greater than indomethacin greater than salicylamide greater than phenylbutazone greater than phenacetin greater than aspirin = salicylic acid, was similar to that reported for their anti-inflammator activity and their ability to inhibit prostaglandin synthesis. The antiproliferative activity of these drugs may, in part, account for their anti-inflammatory and toxic actions in vivo.


The synergistic action of L-glutamate and L-aspartate at crustacean excitatory neuromuscular junctions. 1. When L-glutamate and L-aspartate are simultaneously applied to the excitatory neuromuscular junctions of Maia squinado, they produce an increase in the conductance of the post-synaptic membrane much larger than the sum of conductance effects produced by the individual amino acids alone. 2. An examination of the synaptic noise occurring during this synergistic action reveals that the elementary conductnace events produced by aspartate are suppressed and that normal elementary conductance events produced by glutamate are occurring at an enormously increased rate. 3. It is suggested that aspartate causes this potentiation by inhibiting a system for transmitter inactivation in the region of the post-synaptic receptors and that this system, under normal conditions, prevents the access of externally applied glutamate to the synaptic receptors.


The termination of transmitter action at the crustacean excitatory neuromuscular junction. 1. Excitatory junctional currents (e.j.c.s) and miniature excitatory junctional currents (mine.e.j.c.s) have been followed by recording the focal extracellular potential at excitatory neuromuscular junctions of Maia squinado, the Spider Crab. 2. If L-aspartate (concentrations less than or equal to 1 mM) is present in the saline, the average e.j.c. is prolonged by an increase in the duration of its falling phase. 3. No change occurs in the time course of the probablility of release of quanta in the e.j.c. as determined from the histogram of first quantal latencies. 4. The min.e.j.c. is also prolonged by L-aspartate (concentrations less than or equal to 0-5 mM). The rise time increases slightly, the pre-exponential period of the decay phase almost doubles, and the time constant of the final exponential decay increases to a value larger than the average lifetime of the elementary conductance event produced by L-glutamate. 5. Changes in e.j.c.s and min.e.j.c.s produced by aspartate show a striking similarity to the action of neostigmine on the time course of the vertebrate end-plate current. It is proposed that aspartate exerts its action by blocking a process normally reponsible for clearing the synaptic cleft of transmitter.


Membranous localization and properties of ATPase of rat liver lysosomes. Lysosomes isolated from rat liver were found to have ATPase activity (EC No 3.6.1.3). Subfractionation of the lysosomes revealed a membranous localization of ATPase activity. The enzyme has half maximal activity at 0.2 mM ATP and is inhibited by high concentrations of ATP. The apparent Km for divalent metal is 0.2 mM, and either Ca2+ or Mg2+ give maximal activity. The ATPase activity has latency when lysosomes are isolated from rats treated with Triton WR-1339. This latency may be due to the presence of internalized sucrose because the activity of L fraction lysosomes is much less latent and Triton WR-1339 itself is not inhibitory. The latency of glucosaminidase, a marker enzyme for lysosomes, contrasts with the low latency of the ATPase and points to an ATPase with an exposed active site in intact lysosomes.


Growth rate of cultured Novikoff rat hepatoma cells as a function of the rate of thymidine and hypoxanthine transport. Novikoff rat hepatoma cells were propagated in suspension culture in the presence of 1 micron methotrexate and various concentrations of hypoxanthine (or adenosine plus guanosine) and thymidine and with or without the inhibitor of nucleoside and purine transport, Persantin (dipyridamole). Methotrexate-treated cells failed to replicate and died even if the medium was supplemented with either thymidine or a purine source, but normal replication occurred when both were present. The additional presence of Persantin reduced the rate of transport of thymidine or hypoxanthine and thus their incorporation into the nucleotide pool and decreased the rate of cell replication. The growth rate of the cells was directly proportional to the rate of incorporation of thymidine (in the presence of excess hypoxanthine) or of hypoxanthine (in the presence of excess thymidine) until the normal maximum growth rate was obtained. Normal cell replication in the presence of methotrexate and Persantin occurred only when the medium was supplemented with 500 micron hypoxanthine and 30 micron thymidine. The results illustrate a dependence of the growth rate of mammalian cells on the rate of transport of essential nutrients into the cell.


Divalent cation binding to phospholipids: an EPR study. Divalent cation association to sonicated phospholipid liposomes has been examined with electron paramagnetic spectroscopy. Spectra were obtained suggesting that, in some cases, divalent cations associated with acidic phospholipid head groups are highly mobile. Using the amplitude of its characteristic sextet signal as a measure of free Mn(H2O)+6+, the apparent affinities of cardiolipin and phosphatidylserine for Mn2+ were measured as a function of monovalent electrolyte. Monovalent cations having smaller nonhydrated radii were more effective in displacing Mn from the phospholipids. Under conditions of low divalent cation concentrations, it is shown that the Gouy-Chapman diffuse double layer theory predicts a Mn-affinity (KA) inversely proportional to the square of monovalent salt concentration. Although this relationship was closely obeyed for Mn binding to cardiolipin, the fall-off in KA with added sodium chloride was slower in the cases of Mn binding to phosphatidylserin or phosphatidic acid. When phosphatidylcholine or cholesterol was incorporated into mixed vesicles along with a fixed amount of charged phospholipid, the Mn-binding strength was roughly proportional to the weight fraction of the latter. This result is consistent with: (1) a random dispersal of lipids in the bilayer, and (2) a 1:2 divalent cation-phospholipid interaction.


Differences between resting and insulin-stimulated amino acid transport in frog skeletal muscle. We have compared some features of the resting and the insulin-stimulated uptake of alpha-aminoisobutyrate (AIB) in frog skeletal muscle. We found a substantial difference between the two processes, namely, that resting AIB uptake is Na-independent while the insulin-stimulated fraction of the AIB uptake is Na-dependent. Since the amino acid transport systems in frog skeletal muscle are poorly characterized, we have also surveyed some of their properties. One of the most interesting findings of this survey is that both the uptake and efflux of AIB are inhibited by low concentrations of PCMBS (parachloro-mercury-benzene sulfonic acid 5 X 10(-5) M). In contrast, the carrier mediated transport of basic amino acids is neither inhibited by this mercurial agent nor accelerated by insulin. The action of PCMBS strongly suggests the presence of a critical sulfhydryl group in the amino acid carrier system utilized by AIB. This group is exposed to the outside solution since PCMBS penetrates cell membranes poorly, and in addition its inhibitory actions were reverted by agents that do not penetrate the cell membrane like albumin or glutathione.


ATPase and phosphatase activities from human red cell membranes: I. The effects of N-ethylmaleimide. In human red cell membranes the sensitivity to N-ethylmaleimide of Ca2+-dependent ATPase and phosphatase activities is at least ten times larger than the sensitivity to N-ethylmaleimide of (Na+ + K+)-ATPase and K+-activated phosphatase activities. All activities are partially protected against N-ethylmaleimide by ATP but not by inorganic phosphate or by p-nitrophenylphosphate. (ii) Protection by ATP of (Na+ + K+)-ATPase is impeded by either Na+ or K+ whereas only K+ impedes protection by ATP of K+-activated phosphatase. On the other hand, Na+ or K+ slightly protects Ca2+-dependent activities against N-ethylmaleimide, this effect being independent of ATP. (iii) The sensitivity to N-ethylmaleimide of Ca2+-dependent ATPase and phosphatase activities is markedly enhanced by low concentrations of Ca2+. This effect is half-maximal at less than 1 micron Ca2+ and does not require ATP, which suggests that sites with high affinity for Ca2+ exist in the Ca2+-ATPase in the absence of ATP. (IV) Under all conditions tested the response to N-ethylmaleimide of the ATPase and phosphatase activities stimulated by K+ or Na+ in the presence of Ca2+ parallels that of the Ca2+-dependent activities, suggesting that the Ca2+-ATPase system possesses sites at which monovalent cations bind to increase its activity.


ATPase and phosphatase activities from human red cell membranes: II. The effects of phospholipases on Ca2+-dependent enzymic activities. Treatment of human red cell membranes with pure phospholipase A2 results in a progressive inactivation of both Ca2+-dependent and (Ca2+ + K+)-dependent ATPase and phosphatase activities. When phospholipase C replaces phospholipase A2, Ca2+-dependent ATPase activity and Ca2+-dependent phosphorylation of red cell membranes are lost, while Ca2+-dependent phosphatase activity is enhanced and its apparent affinity for Ca2+ is increased about 20-fold. Activation of Ca2+-dependent phosphatase following phospholipase C treatment was not observed in sarcoplasmic reticulum preparation. Phospholipase C increases the sensitivity of the phosphatase to N-ethylmaleimide but has little effect on the kinetic parameters relating the phosphatase activity to substrate and cofactors, suggesting that no extensive structural disarrangement of the Ca2+-ATPase system has occurred after incubation with phospholipase C.


Active chloride secretion by rabbit colon: calcium-dependent stimulation by ionophore A23187. Addition of Ca ionophore, A23187, to the solution bathing the mucosal surface of descending rabbit colon resulted in a reversal of active C1absorption to active C1 secretion, a twofold increase in short-circuit current and a 40% increase in tissue conductance without affecting the rate of active Na absorption. These alterations in electrolyte transport are quantitatively similar to those previously observed in response to cyclic 3',5'-AMP (cAMP) (RA. Frizzell, M.J. Koch & S.G. Schulz, J. Membrane Biol. 27:297, 1976). When medium Ca concentration was reduced to 10(-6) M, the secretory response to A23187 was abolished but the response to cAMP was unaffected. The ionophore did not influence the cAMP levels of colonic mucosa. Addition of cyclic AMP to colonic strips preloaded with 45Ca elicited a reversible increase in Ca efflux from the tissue. These results suggest that an increase in intracellular Ca concentration stimulates colonic electrolyte secretion and that the secretory response to cAMP may be due, at least in part, to a release of Ca from intracellular stores.


Biochemical studies of taste sensation: II. Labelling of cyclic AMP of bovine taste papillae in response to sweet and bitter stimuli. Labeling of cyclic AMP of tase papillae and its responsiveness to tast stimuli has been measured using whole papillae from bovine tongue prelabeled with [ 8-(14) C] adenine. Labeling was measured in circumvallate and fungiform papillae, both of which contain taste buds, and in filiform papillae and small blocks of tongue epithelium, which are devoid of taste buds. No differences were observed in the levels of activity. The labeling of cyclic AMP of circumvallate papillae showed only small increases (12-22%) in the presence of the taste stimulus sucrose (sweet), and the stimulatory effects were not statistically significant. The increase due to sucrose was not potentiated by theophylline. No stimulation by sucrose was observed with epithelium controls. Lactose, which is a poor taste stimulus, did not stimulate labeling of cyclic AMP in taste papillae. Theophylline, caffeine, and quinine (bitter) stimulated labeling of cyclic AMP by up to 2-fold, as did L-Epinephrine. Evidence for a specific role of cyclic AMP as a second messenger in taste sensation was not obtained. It is suggested that cyclic AMP might provide a mechanistic basis for studying some of the effects of sweet and bitter compounds in mixtures.


Genetic studies of the Fv-1 locus of mice: linkage with Gpd-1 in recombinant inbred lines. Multiple recombinant inbred lines, derived from crosses between strains permissive to N-tropic murine leukemia viruses (Fv-1n) and strains permissive to B-tropic murine leukemia viruses (Fv-1b), have been characterized as to Fv-1 genotype and other chromosome 4 markers, including the closely linked hexose-6-phosphate dehydrogenase isozyme locus (Gpd-1). Only one recombinant between Fv-1 and Gpd-1 was found among 45 lines tested. On this basis, the distance between Fv-1 and Gpd-1 is estimated to be 0.6 centimorgans. None of the lines was either resistant or susceptible to both N- and B-tropic viruses. Nineteen other inbred strains, previously untested, were characterized as either Fv-1n or Fv-1b.


Physiological state of human embryonic lung cells affects their response to human cytomegalovirus. Cultures of human embryonic lung (HEL) cells in different physiological states were studied for their susceptibility to infection with human cytomegalovirus (CMV) with respect to production of infectious virus, synthesis of viral antigens, and virus-induced stimulation of cellular DNA synthesis. In general, subconfluent, actively growing cells yielded higher amounts of infectious virus than did confluent contact-inhibited cells. The higher yield of infectious virus was correlated with a greater percentage of cells producing viral antigens within the first 48 h after infection. In confluent cultures, 25 to 50% of the cells produced viral antigens within the first 48 h postinfection. This proportion did not change over a 10-fold range of multiplicity of infection, indicating that many of the cells in confluent cultures did not support productive infection. However, virtually all the cells in subconfluent cultures were susceptible. Also, in contrast to herpes simplex virus and pseudorabies virus, infectious CMV is not produced by cells treated with 5-fluorouracil and thymidine. Virus-induced stimulation of cellular DNA synthesis in cells infected at high multiplicities of infection could be detected only in confluent cultures, in which cellular DNA synthesis had been previously suppressed, but could not be detected in similarly treated cultures of subconfluent cells. The lack of detectable stimulation of cellular DNA synthesis in the latter was related to the fact that practically all the cells in the culture synthesized viral antigens within the first 48 h after infection, productive infection and detectable synthesis of cellular DNA being mutually exclusive.


Complementation rescue of Rous sarcoma virus from transformed mammalian cells by polyethylene glycol-mediated cell fusion. Polyethylene glycol (PEG) is effective as a fusing agent for the rescue of virus from Rous sarcoma virus-transformed mammalian cells. The procedure of PEG-mediated rescue of virus from virogenic cell lines is described, and the technique is compared with that of Sendai virus-mediated rescue. Virus may be rescued quantitatively from virogenic cell lines by plating mitomycin C-killed transformed mammalian cells with chicken embryo cells, treating the monolayers with 50% PEG and overlaying the monolayers with focus agar. The number of foci that appeared reflected the number of heterokaryons in the fusion mixtures that released infectious virus. PEG gave reproducible results in virus rescue experiments with an efficiency equal to the best Sendai virus preparations. In addition to the description of the technique for PEG-mediated virus rescue from virogenic cell lines, a method for virus rescue from nonvirogenic lines is presented. Preinfection of the chicken embryo cells with helper avian leukosis virus (Rous-associated virus) prior to fusion with mammalian cells transformed by defective viruses complements the virus defect. We examined four nonvirogenic cell lines, and all released infectious virus in the complementation rescue assay.


Oligonucleotide fingerprints of RNA species obtained from rhabdoviruses belonging to the vesicular stomatitis virus subgroup. The relationships among the genomes of various rhabdoviruses belonging to the vesicular stomatitis virus subgroup were analyzed by an oligonucleotide fingerprinting technique. Of 10 vesicular stomatitis viruses, Indiana serotype (VSV Indiana), obtained from various sources, either no, few, or many differences were observed in the oligonucleotide fingerprints of the 42S RNA species extracted from standard B virions. Analyses of the oligonucleotides obtained from RNA extracted from three separate preparations of VSV Indiana defective T particles showed that their RNAs contain fewer oligonucleotides than the corresponding B particle RNA species. The fingerprints of RNA obtained from five VSV New Jersey serotype viruses were easily distinguished from those of the VSV Indiana isolates. Three of the VSV New Jersey RNA fingerprints were similar to each other but quite different from those of the other two viruses. The RNA fingerprints of two Chandipura virus isolates (one obtained from India and one from Nigeria) were also unique, whereas the fingerprint of Cocal virus RNA was unlike that of the serologically related VSV Indiana.


Regulation of early and late simian virus 40 transcription: overproduction of early viral RNA in the absence of a functional T-antigen. Virus-specific RNA synthesized in monkey cells after infection by both wild-type simian virus 40 (SV40) and the early SV40 temperature-sensitive mutant tsA58 has been analyzed. The fraction of SV40-specific RNA increased throughout infection with either wild-type SV40 or with tsA58 in direct proportion to the accumulation of progeny DNA molecules, suggesting their role in the late transcriptional process. Cytoplasmic fractions from cells infected at various temperatures (31.5 to 41 degrees C) by wild-type virus and harvested 48 h later contained 4 to 8% virus-specific RNA, of which 5 to 10% was early SV40 RNA. In contrast, though 5 to 8% of the cytoplasmic RNA from tsA 58-infected cells incubated at 31.5 to 37 degrees C for 48 h was virus specific, the percentage of early virus-specific RNA ranged from 25 to 80% as the incubation temperature increased. In tsA58-infected cultures incubated for 48 h at 41 degrees C (a temperature at which essentially no tsA 58 DNA synthesis occurred), only 0.4% of the cytoplasmic RNA was virus specific, but at least 90% of this RNA was early. In experiments where cells were inoculated at 32 degrees C and shifted at 48 h postinfection to 40 degrees C for various times, the percentage of virus-specific pulse-labeled RNA varied from 3.5 to 10.0%. Of the virus-specific RNA, early SV40 RNA ranged from 14 to 65% in tsA 58-infected cultures. Analogous studies with Sarkosyl-extracted viral transcription complexes to incorporate label into nascent (unprocessed) viral RNA yielded essentially identical results. This finding strongly suggests that the overproduction of early SV40 RNA occurs at the level of synthesis. While cytosine arabinoside effectively terminated most viral DNA replication in wild-type-infected cells, the ratio of early to late viral RNA remained less than 1:9. These results demonstrate that: (1) the amount of virus-specific RNA synthesized depends directly on the amount of viral DNA available for use as templates; once viral DNA replication has occurred, presumably providing progeny SV40 DNA molecules for templates, the level of transcription remains high; (ii) termination of viral DNA replication does not terminate late SV40 transcription; (iii) early SV40 RNA is overproduced by tsA 58 at all temperatures, but especially at higher temperatures; and (iv) overproduction of early SV40 RNA appears to be correlated with defectiveness of the tsA mutant T-antigen. These results suggest that T-antigen may regulate its own production either by repressing the synthesis of early viral RNA or by stimulating the synthesis of late SV40 RNA or both.


Polypeptide synthesis in simian virus 5-infected cells. Polypeptide synthesis in three different cell types infected with simian virus 5 has been examined using high-resolution polyacrylamide slab gel electrophoresis, and all of the known viral polypeptides have been identified above the host cell background. The polypeptides were synthesized in infected cells in unequal proportions, which are approximately the same as they are found in virions, suggesting that their relative rates of synthesis are controlled. The nucleocapsid polypeptide (NP) was the first to be detected in infected cells, and by 12 to 14 h the other virion structural polypeptides were identified, except for the polypeptides comprising the smaller glycoprotein (F). However, a glycosylated precursor (F(0)) with a molecular weight of 66,000 was found in each cell type, and pulse-chase experiments suggested that this precursor was cleaved to yield polypeptides F(1) and F(2). No other proteolytic processing was found. In addition to the structural polypeptides, the synthesis of five other polypeptides, designated I through V, has been observed in simian virus 5-infected cells. One of these (V), with a molecular weight of 24,000, was found in all cells examined and may be a nonstructural viral polypeptide. In contrast, there are polypeptides present in uninfected cells that correspond in size to polypeptides I through IV, and similar polypeptides have also been detected in increased amounts in cells infected with Sendai virus. These findings, and the fact that the synthesis of all four of these polypeptides is not increased in every cell type, suggest that they represent host polypeptides whose synthesis may be enhanced upon infection. When a high salt concentration was used to decrease host cell protein synthesis in infected cells, polypeptides IV and (to a lesser extent) I were synthesized in relatively greater amounts than other cellular polypeptides, as were the viral polypeptides. The possibility that these polypeptides may play some role in virus replication is discussed.


RNase T1-resistant oligonucleotides of B-tropic murine leukemia virus from BALB/c and five of its NB-tropic derivatives. We used two-dimensional gel electrophoresis to obtain fingerprints of RNase T1-resistant oligonucleotides of a B-tropic murine leukemia virus from BALB/c and five NB-tropic viruses independently derived from this B virus by passage through NIH Swiss mouse embryo cells in vitro. The fingerprints of the B- and NB-tropic viruses were very similar: approximately 33 of 35 large T1-resistant oligonucleotides appeared to be shared by these viruses. However, the five NB-tropic viruses possessed an apparently common alteration relative to their B virus progenitor. This change involved the acquisition of one oligonucleotide and, tentatively, the loss of one oligonucleotide. We do not know whether these changes represent an alteration responsible for the change from B- to NB-tropism. Fingerprints of B- and NB-tropic viruses were not affected when the viruses were grown in cells of different Fv-1 type.


Properties of simian virus 40 transcriptional intermediates isolated from nuclei of permissive cells. A nucleoprotein complex that is an intermediate in viral transcription has been isolated from simian virus 40 (SV40)-infected BSC-1 cells after lysing infected nuclei with Sarkosyl. It contain DNA, DNA-dependent RNA polymerase II, and nascent RNA chains. RNA chain elongation continues for several hours in vitro and is dependent on exogenous ribonucleoside triphosphates. The complex sediments in neutral sucrose gradients with a main peak at about 24 to 26S. When the nascent RNA on the complex is treated with RNase A, a fraction of the RNA remains resistant to RNase and is hydrogen bonded to the DNA template. The pulse-labeled RNase-resistant RNA can be chased into RNase-sensitive RNA, indicating that it is located at the 3' terminus of the RNA chain. The rate of RNA displacement from the DNA template is consistent with an average rate of RNA chain elongation of 15 to 30 nucleotides per min. At least 70% of the RNA synthesized in this in vitro system is SV40 specific. Hybridization with the separated strands of SV40 DNA and with fragments of SV40 DNA generated with endonucleases HindII + III indicates that this RNA  is complementary to all regions of the "late" SV40 DNA strand. Studies of SV40 RNA synthesis in this partially purified preparation at early and late times after infection should provide a way of locating promoter sites for transcription and identifying the form of SV40 DNA that serves as a template for late transcription.


Uniform representation of the human papovavirus BK genome in transformed hamster cells. The DNA of three cloned lines of hamster kidney cells transformed by human papovavirus BK DNA was examined by reassociation kinetics for viral sequences and found to contain 2.7 to 5.3 equivalents of viral DNA per diploid genome. In the one line examined with the four R-HindIII fragments of the human papovavirus BK genome, the entire viral genome was uniformly represented.


Ammonium chloride inhibits cell fusion induced by syn mutants of herpes simplex virus type 1. Cell fusion induced by syn mutants of herpes simplex virus type 1 is inhibited by NH4Cl. The inhibition of fusion is both rapid and rapidly reversible and apprears to be due to the NH4+ ion. Virus production was not significantly altered by NH4Cl, although cell growth was greatly diminished.


Type C viral gag gene expression in chicken embryo fibroblasts and avian sarcoma virus-transformed mammalian cells. Sensitive radioimmunoassays were developed for avian type C viral gag gene-coded proteins. These assays were used to examine the restriction to virus production by avian embryo cells and mammalian cells transformed by avian sarcoma viruses. The results indicate that although a high-molecular-weight primary translational product of the gag gene is expressed, its cleavage and processing are incomplete. Furthermore, analysis of intermediate cleavage products provided information regarding the order of sequences coding for the individual viral proteins within the avian type C viral gag gene.


Sizes and concentrations of several type C oncornaviruses and bacteriophage T2 by the resistive-pulse technique. Viruses above about 60 nm in diameter may be rapidly sized to a few nanometers in their natural hydrated state as they pass one by one through a single pore in a newly developed nanometer-particle analyzer based on the resistive-pulse technique of the Coulter Counter and the use of submicron diameter pores made by the Nuclepore process. Size measurements for several type C oncornaviruses are: Rauscher murine leukemia, 122.3 +/- 2 nm; simian sarcoma, 109.7 +/- 3 nm; Mason-Pfizer monkey, 140.0 +/- 2.5 nm; RD-114, 115 +/- 5 nm; and feline leukemia, 127.4 +/- 2 nm, relative to standard 109-nm latex spheres. The T2 bacteriophage has a volume of (5.10 +/- 0.15) X 10(-16) cm3. Concentrations of viruses near 10(9) to 10(11)/ml that are fairly clear of debris are routinely measurable in a few minutes to an accuracy near 15%. A lower practical count limit is near 5 X 10(7) viruses per ml.


5'-Terminal and internal methylated nucleosides in herpes simplex virus type 1 mRNA. RNA labeled with [methyl-3H]methionine and/or [32P]orthophosphate was isolated from the polyribosomes of herpes simplex virus (HSV) types 1-infected cells and separated into polyadenylylated [poly(A+)]and non-polyadenylylated [poly(A-)] fractions. Virus-specific RNA was obtained by hybridization in liquid to either excess HSV DNA or filters containing immobilized HSV DNA. Analysis in denaturing sucrose gradients indicated that HSV-specific poly(A+) RNA sedimented in a broad peak, with a modal S value of 20. The ratio of [3H]methyl to 32P decreased with increasing size of RNA, suggesting that each RNA chain contains a similar sumber of methyl groups. Further analysis indicated an average of one RNase-resistant structure of the type m7G(5')pppNmpNp or m7G(5')pppNmpNmpNp per 2,780 nucleotides. The following components were identified in the 5'-terminal oligonucleotides of polyribosome-associated HSV-specific poly(A+) and poly(A-) RNA: 7-methylguanosine, N6,2'-O-dimethyladenosine, and the 2'-O-methyl derivatives of guanosine, adenosine, uridine, and denosine, and the 2'-O-methyl derivatives of guanosine, adenosine, uridine, and cytidine. The most common 5'-terminal sequences were m7G(5')pppm6Am and m7G(5')pppGm. An additional modified nucleoside, N6-methyladenosine, was present in an internal position of HSV-specific RNA.


Interaction of polyoma and mouse DNAs. IV. Time course and extent of integration of polyoma DNA into mouse DNA during lytic infection. The time course of covalent binding of polyoma viral DNA to mouse DNA was followed in mouse embryo cells that had been grown prior to infection in the presence of 5-bromodeoxyuridine. Density-labeled (HL) mouse DNA was separated from free polyoma DNA by CsCl isopycnic centrifugation. Polyoma DNA sequences present in HL mouse DNA were detected by hybridization with radioactive cRNA synthesized in vitro. In reconstruction experiments, the limit of detection was found to be, on the average, about 0.5 genome equivalent (g.e.) of polyoma DNA per cell. To find conditions for the isolation of HL mouse DNA and for its complete separation from free polyoma DNA, cultures infected at 4 degrees C were used. HL mouse DNA extracted with sodium dodecyl sulfate and high salt concentrations (5 to 6 M CsCl) and then purified by three consecutive CsCl density gradient centrifugations was free from detectable amounts of polyoma DNA, whereas HL mouse DNA extracted with chloroform and phenol and purified in the same way always contained contaminating, noncovalently bound polyoma DNA. In lytically infected bromodeoxyuridine-prelabeled mouse embryo cultures, polyoma DNA bound to HL mouse DNA that had been extracted by the sodium dodecyl sulfate-CsCl procedure was first detected in small amounts (1 to 2 g.e. per cell) at 10 h after infection. In cultures incubated with medium containing thymidine (5 mug/ml), 4 to 6 g.e. of polyoma DNA per cell was detected at 14 and 18 h after infection. In these samples, practically all viral DNA was bound to high-molecular-weight HL mouse DNA. In cultures incubated with normal medium (no additions) and extracted between 17 and 20 h after infection, 20 to 350 g.e. of polyoma DNA per cell banded with HL mouse DNA. However, when DNA of one of these samples was subfractionated by sodium dodecyl sulfate-salt precipitation prior to isolation of HL mouse DNA, about 80% of the viral DNA banding at increased density was present in the low-molecular-weight DNA fraction. This observation suggests that in normal medium some progeny viral DNA of increased density was synthesized. Covalent binding of polyoma DNA to density-labeled mouse DNA was demonstrated by alkaline CsCl density gradient centrifugation: nearly equal amounts of polyoma DNA were found in the H and L strands, respectively, as is expected for linear integration of viral DNA. The results lead to the conclusions that (i) early polyoma mRNA is transcribed from free parental viral DNA; (ii) covalent linear integration is first detectable at the time when tumor (T)-antigen is synthesized; and (iii) only few copies (<10 g.e./cell) become integrated between 10 and 18 h after infection, i.e., during the period when cellular and viral DNA replication starts in individual cells.


Addition of extra DNA sequences to simian virus 40 DNA in vivo. The possible addition of extra sequences to simian virus 40 (SV40) DNA was analyzed by electron microscopy in two different cell systems, productively infected monkey cells and activated heterokaryons on monkey and transformed mouse 3T3 cells. We found that the closed circular DNA fraction, extracted from monkey cells at 70 h after infection with nondefective SV40 at a multiplicity of infection of 6 PFU/cell, contained oversized molesules (1.1 to 2.0 fractional lengths of SV40 DNA) constituting about 8% of the molecules having lengths equal to or shorter than SV40 dinner DNA. The oversized molecules had the entired SV40 sequences. The added DNA was heterogeneous in length. The sites of addition were not specific with reference to the EcoRi site. These results suggest that recombination between monkey and SV40 DNAs or partial duplication of SV40 DNA occurs at many sites on the SV40 chromosome. The integrated SV40 DNA is excised and replicates in activated heterokaryons. In this system, besides SV40 DNA we found heterogeneous undersized and oversized molecules containing SV40 sequences in the closed circular DNA population. Additions differeing in size appeared to be overlapping and to have occurred at a preferential site on the SV40 chromosome. These results support the hypothesis that host DNA can be added to SV40 DNA at the site of integration at the time of excision.


Encephalomyocarditis virus RNA. II. Polyadenylic acid requirement for efficient translation. Differentially polyadenylated subpopulatons of encephalomyocarditis (EMC) viral RNA were isolated by affinity chromatography on oligodeoxythymidylic acid-cellulose. Translation of these RNA fractions in several in vitro protein-synthesizing systems, isolated from Ehrlich ascites tumor cells, demonstrated that poly(A)+EMC viral RNA was translated two to three times more efficiently than poly(A)-EMC viral RNA. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of the polypetides synthesized by the in vitro system in response to the different RNAs showed no detectable differences in the size or relative amount- of the translational products. mRNA saturation curves indicated that the in vitro systems were stimulated maximally by equivalent amounts of RNA, wheter it be poly(A)-or poly(A)+ EMC viral RNA. Time course experiments showed that the differences in translatability were more pronounced late in the reaction when reinitiation was required, and that by eliminating reinitiation with high salt the apparent effect of poly(A) on translation was diminished. Together, these results suggest that poly(A) may be required for efficient initiation and reinitiation of protein synthesis in the cell-free systems. This interpretation is discussed relative to earlier data.


Heteroduplex study of the sequence relations between RD-114 and baboon viral RNAs. The regions of sequence homology and nonhomology between the RNA genomes of RD-114 and baboon endogenous type C viruses have been mapped by an electron microscope heteroduplex study. Short complementary DNA (cDNA) copies (approximately 150 to 200 nucleotides in length) of RD-114 RNA were prepared by an endogenous synthesis; labels of polydeoxythymidylic acid [poly(dT)] were attached to the 3' ends of the cDNA molecules by a reaction catalyzed by deoxynucleotidyl terminal transferase. The cDNA-poly(dT) was hybridized to RD-114 RNA and to baboon viral RNA dimer (50 to 70S) units, and the position- of the poly(dT) labels were observed by electron microscopy. With RD-114, labels were distributed uniformly along the genome. With baboon virus RNA (monomer length, 9.5 kilobases [kb]), the regions of high homology with RD-114 cDNA were observed to lie in the intervals from 1.5 to 2.5 kb and from 3.7 to 5.5 kb from the 5' end. The relations of these heteroduplex maps to the known antigenic similarities and differences among the several viral proteins and to the genetic maps of the viruses are discussed.


Characterization of the protein and nucleic acid of Aleutian disease virus. Aleutian disease virus (ADV) was extracted and purified from infected mink. Nucleic acid extracted from the virus was examined in an electron microscope. Three different sizes of molecule, with approximate lengths of 1.2, 0.55, and 0.25 micron, were observed. The ratios of the large molecules to the small molecules were similar in all the particles prepared under different conditions. Equilibrium CsCl density gradient centrifugation showed that ADV nucleic acid had a buoyant density of 1.733 g/cm3. In Cs2SO4, ADV had a lower buoyant density than that of double-stranded RNA. These properties and its sensitivity to DNase suggested that ADV contains DNA. Thermal denaturation curves revealed that the DNA of ADV had a single-stranded configuration. Polypeptide analysis of ADV by polyacrylamide gel electrophoresis revealed the presence of four polypepties, with molecular weights of 30,000, 27,000, 20,500, and 14,000. These polypeptides were present in a ratio of 10:3:10:1, respectively. The data suggested that ADV is closely related to the members of the parvovirus groups.


Morphogenesis of poliovirus. IV. existence of particles sedimenting at 150S and having the properties of provirion. An apparent precursor to the poliovirion that cosediments with the virion at 150S was identified by its content of VP-0. It has properties previously associated with the provirion, a structure that sedimented at 125S, and it may be an alternate form of provirion. Like virions, the 150S precursor binds to and elutes from cells, after which it sediments at about 125S.


Anatomy of herpes simplex virus DNA VIII. Properties of the replicating DNA. This paper concerns the properties of herpes simplex virus 1 DNA replicating in HEp-2 and human embryonic lung cells. The results were as follows. (i) Only a small fraction of input viral DNA entered the replicative pool. The bulk of the input viral DNA cosedimented with marker viral DNA and did not appear to be degraded or dissociated into L and S components. (ii) Nascent DNA sedimented faster and banded at a higher density than that of mature viral DNA extracted from virions. Pulse-chase experiments indicated that nascent DNA acquires the sedimentation rate and buoyant density of viral DNA within 30 to 40 min after its synthesis. (iii) Electron microscopic studies indicated that the DNA extracted from cells replicating viral DNA and banding at the density of viral DNA contained: (a) linear, full-size molecules with internal gaps and single-stranded regions at termini; (b) molecules with lariats, consisting of a linear segment up to 2x the size of mature DNA and a ring ranging from 0.5 x 10(6) to 100 x 10(6) in molecular weight, showing continuous and discontinuous forks; (c) circular, double-stranded molecules, both full-size and multiples of 18 x 10(6) in molecular weight, but without forks or loops; (d) molecules showing "eye" and "D" loops at or near one end of the DNA; (e) large, tangled masses of DNA, similar to those observed for T4 and pseudorabies virus replicating DNAs, containing loops and continuous and discontinuous forks. The electron micrographs are consistent with the hypothesis that the single-stranded ends on the DNA anneal to form a hairpin, that the DNA synthesis is initiated at or near that end and proceeds bidirectionally to form a lariat, and that resulting progeny derived by semiconservative replication are "head-to-head" and "tail-to-tail" dimers.


Persistent adenovirus infections of nonpermissive monkey cells. Persisten infections of monkey cells have been established by using two serotypes of human adenovirus. The persistently infected cells show no morpho logical changes, but continue ot produce low titers of infectious adenovirus. The inapparent infection can, at any time, be converted to a cytolytic productive one by superinfection with simian virus 40. Persistence in this system does not appear to result from multiple rounds of lytic infection, nor is it mediated by production of defective interfering particles. The persistently infected cells do not possess the characteristics of oncogenic transformation. Results of these studies also whow that the nonpermissiveness of monkey cells to adenovirus replication can be partially overcome by infection at high multiplicity.


DNA of Epstein-Barr virus. II. Comparison of the molecular weights of restriction endonuclease fragments of the DNA of Epstein-Barr virus strains and identification of end fragments of the B95-8 strain. Incubation of the DNA of the B95-8 strain of Epstein-Barr virus [EBV (B95-8) DNA] with EcoRI, Hsu I, Sal I, or Kpn I restriction endonuclease yielded 8 to 15 fragments separable on 0.4% agarose gels and ranging in molecular weight from less than 1 to more than 30 x 10(6). Bam I and Bgl II yielded fragments smaller than 11 x 10(6). Preincubation of EBV (B95-8) DNA with lambda exonuclease resulted in a decrease in the Hsu I A and Sal I A and D fragments, indicating that these fragments are positioned near termini. The electrophoretic profiles of the fragments produced by cleavage of the DNA of the B95-8, HR-1, and Jijoye strains of EBV were each distinctive. The molecular weights of some EcoRI, Hsu I, and Sal I fragments from the DNA of the HR-1 strain of EBV [EBV (HR-1) DNA] and of EcoRI fragments of the DNA of the Jijoye strain of EBV were identical to that of fragments produced by cleavage of EBV (B95-8) DNA with the same enzyme, whereas others were unique to each strain. Some Hsu I, EcoRI, and Sal I fragments of EBV (HR-1) DNA and Kpn I fragments of EBV (B95-8) DNA were present in half-molar abundance relative to the majority of the fragments. In these instances, the sum of the molecular weights of the fragments was in excess of 10(8), the known molecular weight of EBV (HR-1) and (B95-8) DNA. The simplest interpretation of this finding is that each EBV (HR-1), and possibly also (B95-8), DNA preparation contains two populations of DNA molecules that differ in the arrangement of DNA sequences about a single point, such as has been described for herpes simplex virus DNA. Minor fragments could also be observed if there were more than one difference in primary structure of the DNAs. The data do not exclude more extensive heterogeneity in primary structure of the DNA of the HR-1 strain. However, the observation that the relative molar abundance of major and minor fragments of EBV (HR-1) DNA did not vary between preparations from cultures that had been maintained separately for several years favors the former hypothesis over the latter.


Monoplace hyperbaric oxygen therapy for gas gangrene. Untreated gas gangrene is a fulminating infection that can lead to extensive tissue necrosis and death. Hyperbaric oxygen, when used with antibiotics and surgical debridement, can lead to decreased mortality. Nine patients with gas gangrene proven by positive clostridial cultures have been treated at St Luke's Hospital of Kansas City with a mortality of 11.1%. A case of gas gangrene developing from a perirectal abscess is presented.


Protective action of estrogen in endotoxin shock. Protective action of conjugated equine estrogen (CEE) against endotoxin shock was admitted in mice. Male DDN strain mice pretreated intraperitoneally with 2 mg of CEE showed various survival rates according to the interval between administration of CEE and endotoxin injection. Survival rates of the control were 20.4%, but in the group pretreated with CEE they were 0% at 0 hour, 26.3% at 1 hour, 33.3% at 12 hours, 55.7% at 24 hours (p less than 0.001), 10.5% at 36 hours, and 31.2% at 48 hours respectively. Mice pretreated with 2 mg of CEE before 24 hours exhibited excellent resistance to 0.5 mg endotoxin. On the other hand, the hemodynamic effect of CEE was examined in six mongrel dogs, which showed little hemodynamic changes following administration of 10 mg/kg CEE for three hours observation. We speculated that the anti-shock action of CEE was due to the modification of vascular response to vasoactive substances, on the one hand it augmented the vascular contractile response to vasopressors and on the other hand it lessened the hypersensitive vasoconstriction under shock state.


Effect of propranolol on blood pressure and plasma renin concentration in renovascular hypertensive rats. The effect of chronic oral administration of propranolol (39--80 mg/Kg/day, for 21--27 days) on blood pressure, plasma renin concentration, and heart rate were studied in 2 types of renovascular hypertensive rats; one-kidney type with normal plasma renin and two-kidney type with high plasma renin. No significant change was observed in the blood pressure of either model during the administration of propranolol in spite of a significant reduction of heart rate. Plasma renin concentration showed a trend of suppression in two-kidney type rats, but this change was not significant. There was no suppression in one-kidney type rats. These results suggest that the beta-adrenergic system may not play a major role either in regulating plasma renin level or in maintaining the blood pressure in renovascular hypertensive rats.


Enterotoxigenic Escherichia coli diarrhea of travelers: a prospective study of American Peace Corps volunteers. Travelers' diarrhea was studied prospectively in a group of 39 American Peace Corps Volunteers (PCVs) during their first five weeks in Kenya. Twenty-seven developed diarrheal disease and 12 remained well. Multiple episodes were documented in 11 of the symptomatic volunteers. Enterotoxigenic Escherichia coli of many serotypes producing heat-labile and/or heat-stable enterotoxin were isolated from 17 of the 27 volunteers with diarrhea and from 1 of the 12 well volunteers. The enterotoxigenic E. coli were more likely to be antibiotic sensitive than the non-enterotoxigenic E. coli. A serum antibody rise to the heat-labile toxin (LT) was detected in six symptomatic volunteers, five of whom had a positive culture for LT-producing E. coli, and from one asymptomatic, culture negative volunteer. Salmonella cubana was isolated from two volunteers, and three volunteers had serologic evidence of infection with human reovirus-like (rotavirus) agent. This study confirms the role of enterotoxigenic E. coli as a major cause of travelers' diarrhea and suggests that the disease is similar in widely separated geographic areas.


[Drug treatment of hyperlipemia in middle-aged and old persons with ischemic heart disease]. The examination was conducted in 1048 patients with the ischaemic heart disease. Both males, and females displayed most frequently Type II hyperlipidemia. Type IV was more frequent in males, than in females. The incidence of Types IIa and IIb depended on the stage of coronary atherosclerosis, the age and sex of the patients. The concomitant diseases were found to influence the incidence of hyperlipidemia. Among the tested drugs administered in a course of therapy of 4 weeks atromidine proved to be most effective for the examined types of hyperlipidemia (IIa, IIb, IV). Hyperlipidemia relapsed in 1/3 of the patients within 1 year.


[Antiarrhythmic activity of adrenergic blockaders with different mechanisms of action]. Experiments have demonstrated that Alfeprol arrests atrial fibrillation in dogs induced by electric stimulation of the heart, and atrial fibrillation in cats modelled by Akonitine application; it suppresses ventricular tachycardia in dogs occurring following an occlusion of the branches of coronary arteries, and in cats when induced by strophantin intoxication; it prevents fatal ventricular fibrillation in rats poisoned by calcium chloride. The antiarrhythmic effect of Anaprille was not so persistent and manifested itself only in cases of some particular rhythm disorders. Ornid displayed no antiarrhythmic activity. The blocking of adrenergic innervation of the heart by means of beta-adrenergic blockers (Alfeprol, Anaprilline) and sympatholytic (Reserpine, Ornid) considerably increased the animals tolerance of the toxic effect of strophantin, decreasing its arrhythmogenic effect in particular. The mechanisms of the antiarrhythmic effect of antiadrenergic agents are discussed along with the possibilities of their employment for the correction of cardiac sensitivity of glycosides.


[The pathophysiology of recurrent herpes simplex virus disease (author's transl)]. Clinical recurrence of herpes simplex virus (HSV) disease may occur when latent HSV escapes the state of latency and begins again to enter the lytic reproduction cycle. Recent investigation revealed HSV latency in peripheral sensory ganglia and an intra-axonal viral transport between the periphery and the appropriate sensory ganglia. As all previous attempts to demonstrate latent HSV in ocular structures have failed, the neuronal hypothesis for HSV latency and recurrences is the most probable. The consequences for therapy and prophylaxis of HSV disease are discussed.


Glomus jugulare tumor.

Glomus jugulare tumor. A review has been made of the current recommendations for treatment of glomus tumors involving the ear. Thirty cases treated in our clinic have been reviewed, comprised of 8 glomus tympanicum and 22 glomus jugulare tumors. We recommend surgical excision as primary treatment for glomus tympanicum tumors and high voltage radiotherapy as primary treatment for glomus jugulare tumors. Visible residual tumor following adequate radiotherapy may remain unchanged for many years. Some cases that do not show a response to high voltage radiation may be suitable for secondary radical surgical treatment. Fortunately, we have not yet encountered a case of glomus jugulare tumor with nonresponse to high voltage radiation in the doses recommended. Long term follow-up is important.


Surgical management of vagal chemodectomas. Vagal chemodectomas (tumors of the vagal glomus body) are histologically benign and insensitive to radiation therapy. Unless properly managed, these tumors may become life-threatening because of their eventual impingement on surrounding structures, particularly contents of the cranial vault. Patients with chemodectomas of the skull base may be surgically treated without significant functional or cosmetic loss, provided the surgeon possesses the following surgical abilities: 1. temporal bone dissection capabilities; 2. microsurgical skills, including techniques of middle ear reconstruction and neurovascular anastomosis; 3. implementation of simultaneous multiple approaches to the skull base and infratemporal fossa; and 4. the capability of managing technically inherent intraoperative and postoperative problems. Experience from nine patients, all successfully treated surgically, will be summarized.


Alterations in lipid constituents during growth of Mycobacterium smegmatis CDC 46 and Mycobacterium phlei ATCC 354. Phospholipids of Mycobacterium phlei ATCC 354 and Mycobacterium smegmatis CDC 46 consist of cardiolipin, phosphatidyl ethanolamine, tri-acylated dimannophosphoinositide, tetra-acylated dimannophosphoinositide and tetra-acylated pentamannosphosphoinositide. A comparative study of lipid patterns of M. phlei ATCC 354 and of M. smegmatis CDC 46 in relation to age of culture revealed higher total lipid level and increased activity of malate-vitamin K reductase, a phospholipid requiring enzyme, during the early logarithmic growth phase of the former. No appreciable change occurred in the latter. The high total lipid content coincides with an increase in phospholipid, brought about apparently by the increase in malate-vitamin K reductase. Changes in cardiolipin and phosphatidyl ethanolamine appeared to be unique to M. phlei ATCC 354. However, in both bacterial species, a decrease in glyceride and a progressive increase in tuberculostearic acid with a concomitant decrease in oleic acid, occurred with ageing.


Adenosine 3',5'-monophosphate in Mycobacterium phlei and Mycobacterium tuberculosis H37Ra. Adenosine 3',5'-monophosphate (cAMP) is present in slow growing as well as fast growing mycobacteria. Apparently there does not seem to be any direct relationship between either intra- or extracellular cAMP content with the growth rate of bacilli. As compared to that of E. coli grown on a similar energy source, cAMP content is much higher in mycobacteria. cAMP content inside the cells remains unaltered throughout the growth period and this may be due to lack of complete utilization of the major energy source, glycerol. Glucose when added to the cells, suspended in phosphate buffer, caused a remarkable decrease in intracellular cAMP content, a phenomenon well established in other bacteria.


Detection of precursors of quinolinic acid in Escherichia coli. A technique is described which allows the detection of precursors of quinolinic acid produced by Escherichia coli, independent of a bioassay. This is based on a double autoradiogram utilizing radioactive aspartic acid and fructose-1,6-diphosphate (as a source of dihydroxyacetone phosphate). Six radioactive spots are derived from aspartic acid and four are derived from fructose-1,6-diphosphate. The latter four spots correspond to four of the former spots in migration in each of two solvent systems. The significance of these data relative to genetic and enzymatic data is discussed.


Characterization of simian virus 40 transformed African green monkey cells (CV-1). I. Defective virion and viral genome. Several clones of SV40 transformed CV-1 cells have been characterized for the production of T- and V-antigens and for the state of viral genome. The transformed CV-1 cells failed to produce infectious virions as assayed after sonication or cocultivation and fusion with normal CV-1 cells, and were resistant to super-infection by SV40. Some clones of the transformed cells contained V-antigens. The population of V-antigen positive cells varied from 0 to 100% depending on the passage number while the T-antigen positive cells were always 100%. The virions isolated from the transformed cells were similar in morphology to complete SV40, but lighter in density than complete SV40. In one clone, a small amount of SV40 DNA was detectable in a free state while a large proportion of the DNA hybridizable with SV40 3H cRNA was linearly integrated into the cell DNA. The free SV40 DNA was noninfectious, closed circular DNA with a size smaller than infectious SV40 DNA component I. Since the cell extracts of the transformed cells contained an agent(s) which induced T- and V-antigens in normal CV-1 cells, it was suggested that the SV40 transformed CV-1 cells contained free as well as integrated defective SV40 genomes responsible for the synthesis of T- and V-antigens.


Oestrogens and menopausal and postmenopausal women. The majority of women experience a variety of symptoms at the time of the menopause, but these are frequently regarded as being unworthy of management by their doctors. Recent reports of a possible association between exogenous oestrogens and endometrial carcinoma have increased professional reluctance to prescribe oestrogens for menopausal symptoms. This report describes the initial 50 patients who have attended a special clinic established to manage symptomatic menopausal women; common complaints included hot flushes, lack of energy, altered temperament, dyspareunia and headache. Oestrogen therapy was effective in the alleviation of symptoms and the practical aspects of oestrogen use are discussed. It is recommended that with due recognition of its potential complications, oestrogen therapy should be made available to symptomatic menopausal women, and that it requires further study in regard to its place in the long-term prophylaxis of osteoporosis.


[Experimental studies of the effects of Seda-Kneipp on the sleep of sleep disturbed subjects; implications for the treatment of different sleep disturbances (author's transl)]. Seda-Kneipp a compound preparation of valerian and hops was given to sleep disturbed subjects during the second or third of three consecutive nights disturbed by heavy traffic noise. Prior drug administration reduced the noise induced disturbance of sleep stage patterns: slow-wave sleep and stage REM increased. It is recommended that the initial treatment of severe insomnia by "strong" sleeping pills should be followed by a period during which "weak" sleeping pills are given before the drug administration finally is discontinued.


[Therapy of hyperlipoproteinemia type II with Xantinol-nicotinate (author's transl)]. In 61 out-patients with hyperlipoproteinemia type II the action of 50 mg Xantinol-nicotinate per kg body weight on lipids of plasma and lipoproteins was investigated in a three week trial. The results of six patients had been discharged: two patients failed to attend the follow-up examinations, in four patients a lipid lowering effect could not be demonstrated. 40 Type IIa patients and 15 Type IIb patients were examined. In this two groups cholesterol, phospholipids and triglycerides of plasma were significantly lowered. In Type IIa hyperlipoproteinemia mainly the LDL lipids and in Type IIb additionally the VLDL lipids decreased. In Type IIb patients the decrease of lipids were more rapid than in Type IIa patients. This effect is interpreted by the short half life of VLDL. Withdrawel of the drug resulted in an increase of the lipids to the mean values of the untreated patients within ten days.


In vitro metabolism of vitamin D3 by isolated liver cells. Liver cells were prepared from rats fed a rachitogenic diet to investigate the hepatic metabolism of [alpha-1,2-3H2] vitamin D3. Rat hepatocytes suspended in Hanks medium rapidly took up labeled vitamin D3 from the incubation medium and converted this sterol to various metabolites, including 25-hydroxy vitamin D3 (25-OH-D3). There was steady increment in the cellular production of 25-OH-D3 and of the more polar metabolites of vitamin D3 over 3 hr of incubation as determined by thin layer chromatography. Neither the addition of cyclic nucleotides or dexamethasone to, nor the removal of calcium or phosphate from the medium resulted in changes in the rate of conversion of vitamin D3 to its products. Rats pretreated with sodium diphenylhydantoin converted labeled vitamin D3 to its metabolites at the same rate as control rats. These data indicate that isolated liver cells retain the capacity for vitamin D3 hydroxylation, but suggest that the rate of this process does not undergo rapid changes in response to metabolic stimulation.


Activation of rat erythrocyte phosphofructokinase by AMP and by non-physiological concentrations of cyclic AMP. Cyclic AMP (300 micron) activates phosphofructokinase from dialyzed haemolysates of mature rat erythrocytes. The main conclusions are: a) Cyclic AMP, at pH 7.1 and low concentrations of fructose-6-phosphate, is able to reverse the inhibition produced by different amounts of ATP (up to 1.5 mM). b) The cyclic nucleotide is a positive allosteric effector of the enzyme as shown by the displacement of sigmoidal fructose-6-phosphate saturation curve to hyperbolic kinetics in the presence of inhibitory concentrations (1.5 mM) of ATP.c) Cyclic AMP has no significant influence as deinhibitor of phosphofructokinase either at pH 7.1 and non-inhibitory levels (0.25 mM) of ATP or at pH 8.1 and inhibitory (1.5 mM) or non-inhibitory (0.25 mM) concentrations of ATP. Similar conclusions were obtained with 300 micron AMP but not at a lower concentration (3 micron) with both nucleotides. The comparison of cyclic AMP result with those obtained under similar concentrations of AMP suggest that cyclic AMP is really only an "in vitro" modulator of the enzyme from rat erythrocytes, presumably at an AMP regulatory stie, since non-physiological concentrations are required to act as deinhibitor.


Preparation and characterization of isolated parenchymal cells from guinea pig liver. 1. A method is described for the preparation of isolated cells from guinea pig liver. This involved perfusion in situ, in the non-physiological direction, with collagenase. 2. The cell yield was 20--30%, comparable with those from the livers of other species. 3. The ratio of lactate dehydrogenase to glutamate dehydrogenase in the cells was similar to that in vivo, indicating that there was negligible leakage of cytoplasmic enzymes. 4. The concentrations of K+ and adenine nucleotides were initially lower than in the perfused liver; normal values were obtained on incubation, particularly in the presence of substrate. 5. The L-lactate: pyruvate ratio is 16:1, close to established values. The total beta-hydroxybutyrate: acetoacetate ratio indicates that the mitochondrial redox state is more oxidised than in the perfused liver, but the intracellular ratio is similar to that of the intact liver. 6. Rates of gluconeogenesis and ureogenesis, are within the physiological range. Maximal gluconeogeneis from L-lactate was preceded by a lag period. L-lysine stimulated glucose production from L-lactate but did not abolish the lag phase. 7. The effects of aminooxyacetate and octanoate on L-lactate gluconeogenesis were similar to those in the perfused liver.


Changes in the collagenolytic activity released by primary VX-2 carcinoma cultures as a function of tumor growth. Serum-free media of minced tissue cultures of VX-2 rabbit carcinoma contained a specific collagenolytic activity capable of releasing soluble radioactive peptides from [14C]-labeled collagen fibrils. It was also capable of reducing the viscosity of acid-soluble collagen solutions by cleaving the tropocollagen (TC) molecules primarily at one site to TCA (75%) and TCB (25%) fragments. Three chromatographic fractions were separated by gel filtration: F1, (MW 85-110,000) present in larger amounts in early cultures of younger tumor tissue; F2, (MW-35-40,000) the major component with maximum production in the day 3 media of younger and advanced tumor tissues; F3, (MW 18-22,000) the minor component. Early cultures of younger tumor tissue contained a latent collagenase and were subject to trypsin activation suggesting the presence of inactive enzyme precursors or an enzyme-inhibitor complex.


Experimental phenylketonuria: metabolic studies in rat liver. The "in vivo" effects of L-phenylalanine on the gluconeogenic pathway in the liver of fasted rats with experimentally induced phenylketonuria-like characteristics have been investigated. Significant increases of the fructose 6-phosphate, glucose 6-phosphate and glucose concentrations were observed. The study of the effect of L-phenylalanine on the cytoplasmic and mitochondrial redox state and energy charge showed an increase in the mitochondrial NAD+/NADH ratio while the energy charge was virtually unchanged. The effects of phenylalanine and its metabolic derivatives (phenylacetate, phenylethylamine, phenyl-lactate, o-hydroxyphenylacetate and phenylpyruvate) on the activity of lactate dehydrogenase (EC 1.1.1.27), malate dehydrogenase (EC 1.1.1.37) and 3-hydroxybutyrate dehydrogenase (EC 1.1.1.30) in rat liver have been also investigated. Phenylpyruvate inhibited the lactate dehydrogenase activity with a Ki of 5.3 mM. Phenylpyruvate also inhibited both the mitochondrial (Ki = 4 mM) and cytoplasmic (Ki = 5 mM) malate dehydrogenase activities. Phenylpyruvate, phenylacetate and o-hydroxyphenylacetate inhibited the 3-hydroxybutyrate dehydrogenase activity with Ki values of 0.7, 6.0 and 9.5 mM respectively.


Influence of cyclic 3',5'-adenosine monophosphate on uracil uptake by rifampicin treated Escherichia coli cells. Incubation of cells from a wild type strain of E. coli with 0.3 mg/ml rifampicin for 15 minutes lead to a complete inhibition of RNA synthesis measured as the uracil incorporation into the trichloroacetic acid insoluble fraction. In these rifampicin-treated cells [14C]uracil incorporation tended to decrease during a further incubation at 37 degrees. Addition of cyclic AMP increased the inactivation of the system responsible for [14C]uracil uptake. The cyclic nucleotide effect seems to be specific since ATP or 5'AMP did not increase such inactivation.


Incorporation and metabolism of stearic, oleic, linoleic and alpha-linolenic acids in minimal deviation hepatoma 7288 C cells. Minimal Deviation Hepatoma 7288 C cells were cultured in confluent layer with labeled stearic, oleic, linoleic and alpha-linolenic acids. The kinetics of incorporation and conversion to higher homologs was studied. The maximum amounts incorporated in nmoles per mg of cellular protein for stearic, oleic, linoleic and alpha-linolenic acids were 39, 115.6, 90 and 230 respectively. alpha-linolenic acid was converted to octadeca-6,9,12,15-tetraenoic acid (18:1), eicosa-11,14,17-trienoic acid (20:3), eicosa-8,11,14,17 and 5,11,14,17-tetraenoic acids (20:4) and eicosa-5,8,11,14,17-pentaenoic acid (20:5), and also to myristic, palmitic, palmitoleic, stearic and oleic acids. By a mathematical approach, the endogenous pool size of alpha-linolenic acid available for conversion to eicosa-5,8,11,14,17-pentaenoic acid, were calculated. Both values decreased when the cells were preincubated with unlabeled alpha-linolenic acid.


An investigation of the stability of messenger RNAs in cell-free, translational systems from uninfected and mengovirus-infected Ehrlich ascites tumor cells. The stabilities and translation of Ehrlich ascites tumor cell poly(A)-containing mRNA and mengovirus RNA in fractionated cell-free protein synthesizing systems from uninfected and mengovirus-infected Ehrlich ascites tumor cells were studied. During incubation of the systems about 20% of the input RNA is reduced in size and associated with ribosomes engaged in polypeptide synthesis; the remainder is rapidly degraded by RNases. At the end of active translation, both mRNA and nascent proteins are bound to polysomes which are of the same size as those formed during active protein synthesis. The kinetics of protein synthesis closely follow those of RNA hydrolysis. The stabilities of mengovirus RNA and poly(A)-containing mRNA from Ehrlich ascites tumor cells are the same in both systems.


[Myoclonic encephalopathy in childhood (author's transl)]. Two new cases of infantile myoclonic encephalopathy are reported and a survey of literature is given. The disease is characterized by generalised myoclonic jerks in all striated muscles, by cerebellar ataxia and by fast, jerking, mostly conjugated irregular eye movements (opsoclonus). The disease develops mostly during late infancy and early childhood. The pathogenesis is unknown, probably it is caused by immunological reactions to various agents. Treatment with ACTH or corticosteroids leads to rapid remission of the initial neurological symptoms, but it is suggested that therapy does not prevent frequent sequelae of psychomotor retardation and speech distubances. Remarkably, there is the high coincidence of infantile myoclonic encephalopathy and neuroblastoma. Therefore it is necessary to keep in mind the possibility of a causative neuroblastoma in all children with myoclonic encephalopathy and to control repeatly radiological findings and urin-excretion of catecholamines as well as their metabolic products.


[Prognosis of prenatal varicella-zoster-infections in relation to their onset during pregnancy (author's transl)]. 7 children were observed, whose mothers had Varicella-Zoster-Virus-infection. 3 of these children developed postnatal Herpes Zoster without Varicella in their history; 1 child had Varicella in the neonatal period. There are indications that primary infection with Varicella-Zoster-Virus during the first four months of pregnancy may cause deformities in the child; the same during the last five days of pregnancy increases the risk of varicella with a complicated course in the newborn. Independent of the time of infection during pregnancy the prognosis of a secondary Varicella-Zoster-Virus-infection in the mother seems always to be good for the child. Aspects of the child's immunological maturation, still incomplete at birth, are discussed and can help to understand the different prognoses during pregnancy. The importance of serological diagnosis has been emphasized.


[Examination at the bedside (author's transl)]. The difficulties of neurological examination techniques are often over-estimated. It is feared that the large number of individual symptoms cannot be disentangled. But if the principles of classification are known, the individual findings can easily be interpreted and evaluated. If several functions are disordered the disturbance must be sought where the tracts or groups of cells serving these functions lie close together. The investigation of motoricity and sensitivity, the testing of reflexes, coordination, cerebral nerves, circulation in the head and the higher cerebral functions are presented in detail. The significance of the simple clinical examination is considerably increased if the individual symptoms can be seen as an integrated whole.


[Guidelines to the therapie of the most important rheumatic diseases (author's transl)]. Causal therapy of rheumatic diseases is only possible in the few cases of bacterial infection and some arthroses. In the usually etiologically obscure inflammatory forms, however, the considered use of operative, medicinal and physical measures may achieve improvement or long continued remission, even if healing occurs only rarely. Medicinal treatment and the, in principle, equally justified physiotherapy, especially kinesitherapy, are to be planned from the beginning as longterm therapy which demands close cooperation between the fully informed physician and the completely enlightened patient. The high incidence of side effects of the "basis" drugs and the often necessary combinations of palliatives with corticosteroids enforces a strict indication and constant critical appraisal of the current treatment.


Ectopic production of chorionic gonadotropin and its subunits by islet-cell tumors. A specific marker for malignancy. We measured human chorionic gonadotropin (hCG) and its subunits in 76 patients with islet-cell tumors. Seventeen of 27 patients with functioning islet-cell carcinomas had elevated plasma levels of hCG or one of its subunits (hCG-alpha and hCG-beta). Secretion was often discordant; the most frequent finding was an elevated level of hCG-alpha alone. In one patient responding to streptozocin, changes in hCG-alpha correlated with the clinical course. Studies of tumor extracts suggested that the markers observed in the circulation were being produced in the tumor itself. In contrast, none of the 43 patients with benign disease or the six patients with nonfunctioning malignant tumors had elevated levels of hCG, hCG-alpha or hCG-beta. These data show that hCG and its subunits are prevalent and specific markers for islet-cell carcinoma, and suggest that ectopic secretion results from malignant derepression of the genome rather than overproduction by an aberrant "cell rest."


Estrogen and endometrial carcinoma. An independent pathology review supporting original risk estimate. Initial evidence suggested that estrogen therapy increases the risk of endometrial carcinoma. It was then suggested that some studies may have exaggerated the hazard of estrogen therapy by including patients with atypical endometrial hyperplasia among those having endometrial carcinoma. Three internationally recognized pathologists reviewed the histology slides available from the Ziel and Finkle study, which originally reported a risk ratio of 7.6 for estrogen users. At least one of the pathologists concurred with the original diagnosis in all but one case. Furthermore, all pathologists aggreed that 74 per cent (66/89) were correctly diagnosed. In the 66 patients with unanimous diagnosis, 61 per cent (40/66) had used conjugated estrogens, versus 57 per cent (54/94) in the original study. On the basis of 66 patients and 132 matched controls, the revised risk-ratio estimate is 8.1 (with a one-sided 95 per cent lower confidence limit of 4.5), validating the original estimate.


A cluster of Epstein-Barr-virus-associated American Burkitt's lymphoma. We investigated four patients in whom histologically confirmed Burkitt's lymphomas developed during a one-year period. They were young adults living within 50 km of each other in a rural area of Pennsylvania. Two patients were related by marriage, but there was no contact with or between the other two. Although the majority of American Burkitt's lymphomas are not associated with the Epstein-Barr virus, all four patients showed spectra and titers of antibodies to antigens related to that virus that were characteristic of virus-associated Burkitt's lymphoma in Africa. Viral DNA and numerous cells with virus-determined nuclear antigen were found in tumor specimens available from three patients. These cases are unusual, since time-space clustering of Burkitt's lymphoma associated with Epstein-Barr virus has been observed thus far only in Africa.


Annual discourse--don't eat the quails.

Annual discourse--don't eat the quails. Physicians were the first individuals recorded by name in history. Their attempts to define disease influenced fundamentally the cultures and religions of the world. Surgical skills, although highly developed in ancient times, appear to have been less well documented historically than medical disorders. Because of the greater threat to survival that diseases posed, they became incorporated into religious customs. Contagious diseases also influenced greatly the laws, traditions and historical events of the Bible. Leprosy provided a physical example that presumably represented an image of sin, but it probably was no more prevalent as a disease during Biblical times than at present. Many of the Biblical stories assumed to be allegorical may have been founded on medical fact. For example, it appears likely that the quails that poisoned the wandering Jews were contaminated with cyanide.


The location of the polio genome protein in viral RNAs and its implication for RNA synthesis. Evidence is presented that a small protein (VPg) is covalently attached to the 5'-terminal oligonucleotide VPg-pU-U-A-A-A-A-C-A-Gp of the polio genome, to nascent strands of the polio replicative intermediate and to poly(U) of minus strands. A model of polio RNA replication is proposed implicating VPg in initiation of RNA symthesis, possibly as primer.


Experimental studies on vertical infection of mice with Japanese encephalitis virus. II. Effect of inoculation route on placental and fetal infection. Studies were made on the effect of the route of inoculation on the establishment of experimental vertical infection of mice with Japanese encephalitis virus. Mice of the three strains, CFW, C3H/He, and C57BL/6, were inoculated with a field strain of the virus by the intravenous, intraperitoneal, or subcutaneous route. An attempt was made on them to recover the virus from the placenta and fetus. As a result, there were differences in rate of plancental and fetal infection among the route of inoculation. It was dependent the establishment of vertical infection. In general, the rates of placental and fetal infection were the highest in the case of intravenous inoculation, the second highest in the case of intraperitoneal inoculation, and the lowest in the case of subcutaneous inoculation, although there were small differences in this tendency among the mouse strains used. The differences in the rates of placental and fetal infection were presumed to be derived not from the difference in the severity of infection of the respective mothers, but from the degree of direct approach of virus to the placenta as one of the factors.


Antibody estimation by indirect complement fixation test for foot-and-mouth disease in cattle. Antibody against foot-and-mouth disease (FMD) virus was measured by the indirect complement fixation (ICF) test. For this test serum samples were collected from cattle experimentally infected with FMD virus of O, A and Asia 1 types, as well as cattle infected in the field. Two types of antigen were used. One was antigen derived from infected lingual epithelial culture prepared by Frenkel's method with each type of the virus. The other was antigen derived from the lingual epithelium of cattle infected by virus inoculation. ICF antibody began to be dectected about 4 5 days after inoculation. It reached a maximum titer 10  14 days after inoculation, remaining at this titer for about a week or two, and then decreased gradually. It was, however, detectable even 63 days after inoculation. The rise and fall of ICF antibody was parallel with that of neutralizing antibody, although that antibody was always lower in titer than this. ICF antibody was detected type-specifically from cattle infected experimentally and naturally. These results indicated that the ICF test was available for the routine serological diagnosis and epizootiological investigation and research.


Solid phase radioimmunoassay for cyclic AMP using staphylococcal protein A-antibody adsorbent. A radioimmunoassay for cyclic AMP has been developed using protein A containing staphylococci as an immunoabsorbent. Protein A containing heat-killed staphylococci (Cowan I) are coated with rabbit antiserum raised against the 2'-O-succinyl derivative of cyclic AMP coupled to human serum albumin. After washing with a Tween 20 containing buffer, antibody coated staphylococci are diluted with heat-killed staphylococci devoid of protein A (staphylococcus epidermidis) and mixed with [125I]-2'-O-succinyl cyclic AMP tyrosine methyl ester, standards or unknowns. At the end of the incubation, separation of bound and free labelled antigen is achieved by bound and free labelled antigen is achieved by centrifugation. The results are comparable to those obtained with a precipitation assay using polyethylenglycol 6000. Acetylation prior to radioimmunoassay increases sensitivity about 80-fold. 50% depression of zero dose binding occurs at 15--16 femtomoles acetylated cyclic AMP. The crossreactivity with cyclic GMP, ATP, ADP, 5'-AMP and adenosine is extremely low. The present technique is an attractive alternative to the second antibody method or polyethylenglycol precipitation.


[Case of amyloid polyneuropathy].

[Case of amyloid polyneuropathy]. The authors report a case of sporadic secondary amyloidosis with polyneuropathy in a patient aged 50 years. The disease developed during chronic pyelonephritis. The case is described in view of its rare occurrence, diagnostic difficulties and interesting histological findings.


Lack of detectable secretion of ACTH from pituitary homografts of pars intermedia. Hypophysectomized rats bearing grafts of the pars intermedia (PI) in the kidney capsule for 20 days did not show adrenal weights significantly different from those of hypophysectomized controls. Plasma corticosterone was undetectable in the grafted rats, even after the injection of lysine-vasopressin or histamine. On the other hand, MSH activity was present in measueable amounts, independent of the drug administered, in the plasma of the grafted rats. These results suggest that PI transplants do not have the ability to release ACTH, even though it has been previously reported that they contain this hormone.


Cerebrospinal fluid norepinephrine alterations during electrical stimulation of cerebellar, cerebral surfaces in epileptic patients. Lumbar cerebrospinal fluid norepinephrine concentrations were determined by radioenzymatic assay in five epileptic patients receiving double-blind cerebellar stimulation and in three epileptic patients with subdural cerebral surface electrodes. Mean CSF norepinephrine levels were significantly elevated by chronic cerebellar stimulation and significantly depressed after intermittent cerebral cortical stimulation. Lumbar CSF cyclic adenosine monophosphate levels determined by radioimmunoassay were not significantly altered by either cerebellar or cerebral surface stimulation. Our study suggests that (1) electrical stimulation of the anterodorsal cerebellum in man evokes alterations in noradrenergic metabolism. Cerebellar stimulation-induced elevations in norepinephrine may inhibit cerebellar, cerebral, and spinal neuronal activity. In addition, (2) noradrenergic responses to brain surface stimulation may exhibit regional specificity, and (3) noradrenergic alterations evoked by cerebral surface stimulation may not mimic those induced in isolated brain preparations.


Distal myopathy: electron microscopic and histochemical studies. This report describes the clinical, laboratory, and muscle biopsy histochemical and electron microscopic studies of one inherited and two sporadic cases of distal myopathy. Histopathologic and histochemical studies showed numerous myopathic alterations and no significant evidence of denervation. Electron microscopic studies showed a broad spectrum of nonspecific alterations similar to those in other forms of muscular dystrophy. Autophagic vacuoles were prominent in all cases. The inherited case was characterized by an unusual focal granular degeneration that, ultrastructurally, was composed of homogeneous fine granules devoid of other organelles or myofilamens.


In vitro-labeled DNA for detecting viral genomes in multiple sclerosis: I. Papovaviruses. Papovaviruses appear to be neurotropic and one, JC virus, is implicated as the cause of one type of demyelinating disease, progressive multifocal leukoencephalopathy. To investigate whether human papovaviruses play a role in multiple sclerosis, radioactively labeled DNA from BK virus, human papilloma virus, and simian virus 40 was used as a probe in order to detect related unlabeled DNA sequences in DNA isolated from multiple sclerosis brain and/or spinal cord. Labeled viral probes were denatured and DNA allowed to reassociate in the presence of excess unlabeled DNA from multiple sclerosis tissue or from controls. The reassociation rate of the probe is proportional to the concentration of viral DNA present, and an increase in the reassociation rate of the probe over that of control reactions would indicate the presence of unlabeled viral DNA in multiple sclerosis cellular DNA. However, addition of DNA derived from multiple sclerosis patients did not increase rates of reassociation of viral probes. Known human papovaviruses probably have no role in the pathogenesis of multiple sclerosis.


Duchenne dystrophy: ultrastructural localization of the acetylcholine receptor and intracellular microelectrode studies of neuromuscular transmission. Despite focal degeneration and simplification of the postsynaptic region in Duchenne dystrophy, the postsynaptic acetylcholine receptor (AChR) is preserved. This is in contrast to myasthenia gravis where similar postsynaptic alterations are invariably associated with a marked decrease in AChR. There is no extrajunctional spread of AChR in Duchenne dystrophy. The amplitude and frequency of miniature end-plate potentials and the number of transmitter quanta released by nerve impulse are normal but the resting membrane potential is lower than normal. The findings indicate that the release and the postsynaptic responsiveness to acetylcholine are intact in Duchenne dystrophy.


Bone scanning and diagnosis of reflex sympathetic dystrophy secondary to herniated lumbar disks. In two cases, reflex sympathetic dystrophy (RSD) was due to herniated intervertebral disk at L4-5. In both cases, bone scanning with technetium 99m pyrophosphate was of value and its use is recommended in diagnosing mild or early cases of RSD. RSD may be the cause of persisting pain after disk surgery.


Nasopharyngeal tumor initially manifested as myofascial pain dysfunction syndrome. A patient with facial pain of 1 1/2 years' duration, associated with limitation of opening of the mouth, click, and osteodegenerative changes of the temporomandibular joint, was initially thought to be suffering from a myofascial pain dysfunction syndrome. At first, the patient reacted favorably to muscle exercises and an antidepressive drug, but reduced lacrimation and the development of deafness on the affected side led to re-evaluation and a diagnosis of nasopharyngeal tumor. Biopsy confirmed the presence of an adenocarcinoma of the nasopharynx. The variability of symptoms and the diagnostic problems presented by this tumor are discussed.


Knee maturation as a differentiating sign between congenital rubella and cytomegalovirus infections. Knee epiphyseal maturation is retarded in most neonates with congenital rubella infection, whereas neonates with congenital cytomegalovirus infection have relatively normal maturation. Asse-sment of knee maturation is useful in differentiating rubella from cytomegalovirus infection in the neonate.


Tumors of the chest wall in infants and children. Chest wall tumors of children may arise from subcutaneous tissues, the bony thorax, or extrapleural region. Review of experience at our institution and the literature indicates that primary bony tumors are most often malignant; Ewing's sarcoma is the most frequent in this category. Extrapleural tumors are rare but also usually malignant. Clinical and roentgen features of these lesions are discussed.


Epithelial cells and Von Gierke's disease. Epithelial cells and not fibroblasts from human liver and amniotic fluid contain inducible glucose-6-phosphatase (G-6-Pase) activity. The diagnosis of Von Gierke's disease has been made in a patient with hepatomegaly utilizing cultured epithelial cells grown from a liver biopsy. G-6-Pase activity in epithelial cells from this patient could not be induced by dibutyryl cyclic AMP and theophylline. This is the first use of epithelial cells for diagnosis of a metabolic disease. G-6-Pase activity in cloned epithelial cells from amniotic fluid increases 2- to 3-fold after 24-hr exposure to dibutyryl cyclic AMP and theophylline. The prenatal diagnosis of Von Gierke's disease may be possible in a laboratory experienced with these techniques if epithelial cell growth is obtained from amniotic fluid.


Spatial summation effects on lingual vibrotactile thresholds. Ascending lingual vibrotactile thresholds at 250 Hz were obtained for four groups of normal-speaking adult subjects. Contactor areas were varied for each respective group. Threshold values in microns plotted as a function of the area of vibratory displacement indicated a spatial summation function for the larger contactors (greater than .02 cm2). Findings are discussed in terms of the nature of lingual-tactile mechanoreception and possible implications for neural control mechanisms subserving speech production.


Changes in rem dream content during the night: implications for a hypothesis about changes in cerebral dominance across rem periods. REM dream content was scored for categories suggesting the predominant influence of the left hemisphere, e.g., good ego functioning, verbalization, or the right hemisphere, e.g., music, spatial salience, bizarreness. Data from 5 samples of college men showed consistent evidence of an increase in the prominence of left-, but not right-, related categories from earlier to later REM periods. These data suggest there is an increase in left hemisphere control/dominance across the REM periods during the night. Two sets of predictions based on this hypothesis (using more direct estimates of the hypothesized change) yielded supportive evidence. First, as predicted, there was a positive relation between change in percentage of right eye movement (R%) and (a) temporal position of the REM period and (b) change in left-related categories; greater R% was associated with later REM periods and with more prominent left- (but not right-) hemisphere categories. Second, as predicted, there was a positive relation between the diminution of the ratio of left to right EEG amplitudes (L/R) and (a) temporal position of the REM period and (b) prominence of verbal activity. As expected, this relation was attenuated for those subjects showing a preference for left-handedness. Two possible explanations for the inferred increase in left-hemispheric influence during the night are suggested.


Intra- and extracellular measurements of frog neuromuscular transmission upon stretch of the muscle at different stimulus frequencies. Flexible intracellular micro-electrodes were used to study the effect of changes in muscle length on the end-plate potential in the isolated m. cutaneus pectoris for different frequencies of stimulation (1/60-5Hz). A 20% step-wise increase in muscle length within the physiological range increases the end-plate potential immediately by about 50% (range 0-120%) at all frequencies tested. At stimulus frequencies lower than 1/5 Hz this increase is sustained during a period of 15 min stretch. At 1 Hz, however, the initial increase in the end-plate potential amplitude on the average declines within a few minutes to a steady-state value about 35% higher than the steady-state end-plate potential before stretch. At 5 Hz, the initial amplitude increase is followed by a decline of about 15 min duration and the final amplitude is not increased in comparison with the pre-stretch amplitude. The amplitude of the compound muscle action potential of the gastrocnemius muscle with intact circulation shows a similar time dependent increase upon stretch at different stimulus frequencies. It is concluded that stretch of a frog muscle gives both an immediate and a sustained increase in transmitter release from the nerve terminals during prolonged stimulation at frequencies up to about 5 Hz. This effect of stretch on transmitter release can improve in vivo neuromuscular impulse transmission.


The use of a standardized teaching program in diabetes education. A standardized plan for the patient with diabetes has been implemented in our institution to structure the methods and content of diabetic education. To meet the specific needs of each diabetic patient the plan must be individualized by the primary nurse caring for the patient. The plan requires frequent evaluation and revision to update the content and materials used. In our experience, the use of a standardized program for diabetes education has provided primary nurses with a systematic approach to this important facet of nursing care.


Two kinds of opiate receptor.

Two kinds of opiate receptor. Studies with azidomorphine derivatives have revealed that some of them, particularly N-cyclopropylmethylnorazidomorphine (CAM), stimulate some opiate receptors, while inhibit the others. The opiate receptors stimulated by CAM are called opiate A receptors, while those antagonized by CAM are called opiate B receptors. Opiate receptors are located at nerve terminals and upon stimulation decrease the release of a neurotransmitter. Opiate A receptors are most probably located at cholinergic nerve terminals, are present in the guinea pig ileum, mouse vas deferens and in the brain. Their stimulation leads to constipation and mental clouding. Opiate B receptors located on adrenergic nerve terminals are present in the cat nictitating membrane and in the brain. Their stimulation produces analgesia, depression of coughing and respiration, catalepsy, and mental clouding.


The influence of haloperidol and propranolol on behavior and biochemical changes in the brain of mice treated with LSD. The influence of haloperidol and propranolol on aggressiveness, motility exploration and general behavior, and the activity or level of adenylate cyclase, cyclic AMP, and protein kinase in the brain of mice treated with LSD was tested. Haloperidol evidently, and propranolol slightly less, inhibited the behavioral and biochemical changes induced by LSD. It is suggested that psychotomimetic effects of LSD depend on complex action of this compound on aminergic receptors in the central nervous system, and the antipsychotic effectiveness of haloperidol and propranolol is related to interaction of these drugs and LSD with the receptors for monoamines participating in the central neuromediation.


The cellular structure of bronchial carcinoids. A histological and electronmicroscopic study of bronchial carcinoids indicated that these tumours are argyrophilic in keeping with their occurrence in tissue of embryologically foregut origin. Normal bronchi contain cells that resemble, in their electronmicroscopic content, intestinal Kultschitzky-type cells. Characteristic of the carcinoid cell is the presence of intracellular neurosecretory granules seen on electronmicroscopy despite the apparent absence of argentaffin granules under light microscopy.