Effects of the ionophore X-537A on acetylcholine release at the frog neuromuscular junction.

1. X-537A is an ionophore that can carry cations across cell membranes. We studied its effects on spontaneous and stimulated quantal acetylcholine (ACh) release at the frog neuromuscular junction. 2. When neuromuscular transmission was blocked with high Mg2+ or with curare, X-537A markedly increased the end-plate potential (e.p.p.) amplitude. Then a few minutes later the e.p.p. disappeared. 3. When neuromuscular transmission was blocked with hypertonic saline solution, X-537A did not increase e.p.p. amplitude; it did produce many transmission failures. 4. X-537A decreased the depolarization of the end-plate produced by iontophoretically applied ACh. this may account in part for the disappearance of the e.p.p. in solutions containing the ionophore. 5. X-537A depolarized muscle fibres by about 15 mV. 6. When the extracellular divalent cation concentration was very low, X-537A had little or no effect on miniature end-plate potential (min.e.p.p.) frequency. 7. When a divalent cation was present in the extracellular fluid, X-537A increased the frequency of the min.e.p.p.s. The sequence of effectiveness of the divalent ions we have tested is: Ba2+ greater than Sr2+ greater than Ca2+ greater than Mn2+ congruent to Co2+ congruent to Ni2+ greater than Mg2+. There is a rough parallel between these results and the reported affinity of X-537A for various divalent ions. 8. The increase in min.e.p.p. frequency caused by X-537A was transitory, following the increase min.e.p.p. frequency fell to a very low rate or to zero. Then nerve stimulation did not cause quantal release. A second application of X-537A was without effect. 9. X-537A decreased min.e.p.p. amplitude, in accord with the effect on the sensitivity of the end-plate to ACh. 10. The results support the idea that increases in intracellular divalent cation concentrations trigger quantal release from nerve terminals and are involved in the disensitization of end plate receptors to ACh.


The role of cyclic 3',5'-adenosine monophosphate (cyclic AMP) in the ability of sympathetic nerve stimulation to enhance growth and secretion in rat salivary glands in vivo.

1. The cyclic AMP content of the rat salivary glands was measured after sympathetic and parasympathetic nerve stimulation, and after the administration of isoprenaline and phenylephrine. 2. Stimulation of the sympathetic nerves (20 Hz, 1 msec supramaximal voltage for 30 sec of every min) initially raised the cyclic AMP level in both the parotid and submaxillary glands. The rise, which was blocked by propranolol, was not maintained and declined during stimulation from the maximum (reached after 3 min stimulation) to control levels after approximately 15 min. Stimulation of the parasympathetic nerves for 5 min did not raise the cyclic AMP levels. 3. Isoprenaline and phenylephrine raised the cyclic AMP level in both glands. 4. Theophylline enhanced the growth of the parotid and submaxillary glands as measured either by an increase in the wet and dry weights or in acinar axes lengths. N6O2-dibutyryl cyclic monophosphoric acid (dibutyryl cyclic AMP), did not enhance the growth of the glands. 5. It is concluded that cyclic AMP does not directly control cell growth in the rat salivary glands but may be a 'trigger' for events leading eventually to increased growth.


Synaptic depression related to presynaptic axon conduction block.

1. The depression of synaptic transmission, which occurs during prolonged repetitive activation, was examined in the opener muscle of the crayfish walking leg. 2. Excitatory post-synaptic potentials (e.p.s.p.s) initially facilitated but then declined to low amplitudes after about 4000 stimulus pulses had been delivered; this depression is presynaptic in origin; 3. Axon conduction blocks occured at points of bifurcation along the entire length of the presynaptic nerve. This resulted in failure of the nerve impulse to invade some branches of the terminal arborization. 4. Nerve terminal invasion failure caused either intermittent or complete inactiviation of some synaptic release sites; this was associated with depression of the post-synaptic response. 5. The statistics of transmitter release during prolonged repetitive stimulation were examined by focal extracellular recording methods. Transmitter release could be described by binomial statistics, and depression involved a drop in m, n and p. 6. The rate of spontaneous quantal release did not decrease, however, arguing against transmitter depletion. 7. It is concluded that repetitive stimulation eventually leads to depolarization of the axon membrane. This causes impulse propagation failure which reduces the number of synaptic release sites that are activated and mimics a drop in the effective stimulation rate; both effects cause synaptic depression.


Adverse effects of tris hydrochloride, a commonly used buffer in physiological media.

1. Tris 10mM, produced inhibition of motor responses to adrenergic motor nerve stimulation in all smooth muscles studied (rat anococcygeus and vas deferens perfused rabbit ear artery). Up to 30 mM Tris was without significant effect on those motor responses to cholinergic nerve stimulation examined (guinea-pig ileum). 2. a similar reduction in responses to endogenous motor agonists was seen only in the anococcygeus, while in the rabbit ear artery, responses to noradrenaline (NA) were potentiated. 3. tris consistently produced increased tone in the rat anococcygeus and rabbit aorta, but in no other tissue, Small reductions in tone were seen with Tris in spiral strips of rabbit ear artery. 4. inhibitory responses to adrenergic nerve stimulation and to NA in the rabbit jejunum and to inhibitory nerve stimulation in the rat anococcygeus were unaffected by Tris. 5. in the perfused rabbit heart Tris produced a negative inotropic effect and reduced responses to vagal stimulation. Responses to sympathetic nerve stimulation and to ACh and NA were not significantly altered. 6. up to 40 mM Tris had little effect on responses of the rat diaphragm to motor nerve stimulation. 7. the effects of Tris on the rat anococcygeus were not reduced by increased calcium levels but were reduced at lower incubation temperatures. The negative inotropic effect of Tris on the rabbit heart was also reduced at lower temperatures. 8. the buffer HEPES produced a reduction in responses to stimulation in the rat anococcygeus and vas deferens along, but at higher concentrations than those of Tris. Increases in tone in the anococcygeus and aorta were never seen with HEPES. 9. These results indicate that Tris in concentrations commonly used as a buffer in physiological salines can exert toxic effects on neuro-muscular transmission in smooth and cardiac muscle though not in skeletal muscle. The effects are variable, mainly presynaptic and appear to affect in particular motor and especially adrenergic transmission. They may be associated with intracellular metabolic actions of Tris.


Interaction between inhibitory and excitatory synaptic potentials at a peripheral neurone.

1. The interaction between inhibitory and excitatory synaptic potentials in neurones lying in the submucous plexus of guinea-pig ileum has been examined. 2. It was found that during an inhibitory conductance change, electrotonic potentials were more depressed in amplitude than were excitatory synaptic potentials. 3. It is suggested that inhibitory conductance changes may have only a slight effect on the impedance seen by excitatory synaptic currents as much of the excitatory synaptic current flow is likely to be capacitive. 4. A part of the depression of excitatory synaptic potential amplitude was not associated with changes in electrical properties of neurones and it is suggested that inhibitory transmitter may reduce the release of excitatory transmitter.


Statistical fluctuations in charge transfer at Ia synapses on spinal motoneurones.

1. Net inward charge is calculated for unitary Ia e.p.s.p.s. evoked in spinal motoneurones. Fluctuations in this charge are used to describe the fluctuations in transmission at the Ia synapse. 2. The statistical details of variation in transmission at Ia synapses are largely obscured by noise, arising from spontaneous activity and recording apparatus. 3. A computational procedure is described, which utilized the measured statistical properties of the recording noise, and the measured charge histogram of the evoked e.p.s.p. (with noise) to calculate the variations in charge due to the e.p.s.p. alone. 4. The fluctuation in charge transmission at Ia synapses are, in general, non-quantal. Nor are they described by binomial or Poisson statistics. 5. It is suggested that the basic mode of transmission at a single terminal is all-or-none. The fluctuations result from the combined effect of failures at several terminals arising from a single afferent fibre. This hypothesis is supported by further analysis of these results in a subsequent paper (Edwards, Redman & Walmysley, 1976a).


Non-quantal fluctuations and transmission failures in charge transfer at Ia synapses on spinal motoneurones.

1. The origins of fluctuations in charge transfer during the generation of Ia e.p.s.p.s have been investigated. The discrete components which make up the fluctuating e.p.s.p. have been separated. 2. Some e.p.s.p.s fluctuate between two different amplitudes and time courses. These fluctuations have been analysed to show that charge transmission always occurs at one synaptic location, but not always at a second synaptic location. 3. The failures in transmission were study by stimulating the afferent fibre at different frequencies. Although different probabilities of failure were obtained at different frequencies, there was no systematic change in probability with increasing frequency. 4. Single afferents were tetanized and histograms of charge transfer computed during post-tetanic potentiation (p.t.p.). Only half of the units studied showed any p.t.p. In those that did, evidence was found for a decrease in the probability of failure during potentiation. 5. The results could not be used to distinguish between failure of the impulse to always propagate into the terminals, and failure of the terminals to release transmitter following adequate depolarization. 6. The fluctuations in transmission at a single synapse can be described by a binomial process with n = 1 and p less than or equal to 1. Junctional mechanisms consistent with this description are discussed. Alternative mechanisms which associate failures with failure of impulse transmission at afferent fibre branch points are also suggested.


The effect of polarizing currents on unitary Ia excitatory post-synaptic potentials evoked in spinal motoneurones.

1. Depolarizing and hyperpolarizing currents were applied to motoneurones in which unitary Ia e.p.s.p.s were evoked. The results concentrate on those e.p.s.p.s which had time courses which were compatible with somatically located synapses. 2. No reversal of these e.p.s.p.s was observed. Depolarizing currents up to 150 nA simply reduced the peak amplitude. 3. Hyperpolarizing currents caused little, if any, increase in the peak amplitude of the e.p.s.p. The time course of decay became briefer as the membrane was hyperpolarized. 4. Changes in decay time course of the e.p.s.p. which accompanied depolarization and hyperpolarization could be attributed to changes in membrane conductances, rather than to changes in synaptic current time course. 5. The failure of the e.p.s.p. to increase with hyperpolarization was shown to be due to the failure of the synaptic current to increase, rather than to the shunting of anomalous rectification. 6. Chemical and electrical transmission are evaluated against these results and those of the preceding papers.


Blood pressure and sleep in the rat in normotension and in neurogenic hypertension.

1. Blood pressure was recorded in normotensive rats (114 +/- 2 mmHg) and in those made hypertensive by baroreceptor denervation (135 +/- 4 mmHg) during natural sleep. The different states of the wakefulness-sleep cycle were identified by analysis of the frontal electrocorticogram, the cervical electromyogram and by behavioural changes. 2. During synchronized sleep the arterial pressure in the control animals usually remained unchanged as compared to values prevailing when the rats were awake and quiet. The blood pressure usually attained higher levels in the sino-aortically denervated rats during this state. 3. Coincidently with the short episodes of electrocortical desynchronization of the synchronized sleep the blood pressure exhibited rapid oscillations. These fluctuations were greater in the rats with neurogenic hypertension where there were large pressure drops. 4. During desynchronized (REM) sleep arterial pressure underwent marked oscillations. While in the control rats the pressure usually rose, the baroreceptor-denervated rats showed a severe reduction in pressure. Arterial pressure in denervated animals became lower than that of the control rats, 106 +/- 2 and 121 +/- 2 mmHg, respectively. 5. We interpret these results to mean that in the rat alterations in the sensitivity of the baroreceptor reflex could be an important factor in preventing the arterial pressure from rising to hypertensive levels during syndhronized sleep and from dropping to low values during desynchronized (REM) sleep.


[The ACTH of the pituitary pars intermedia].

The present investigation suggests that the intermediate lobe contains corticotrophic cells that are morphologically similar to those of the adenohypophysis. It can also be concluded that the ACTH molecules extracted from the intermediate lobe do not differ from those from the adenohypophysis. The immunocytochemical studies confirm the presence of ACTH in the intermediate lobes of all species studied so far, and stimulate the question of the mechanism of ACTH synthesis at this level. The interrelationships between different peptides of the intermediate lobe, including LPH, ACTH, CLIP and MSH, require further investigations. The intermediate lobe synthesizes and releases ACTH in vitro. This release can be stimulated by crude extracts of the median eminence or of cerebral cortex, and does not seem to be controlled by the CRF. Although ACTH of the intermediate lobe intervenes in vivo in the response to a neurogenic stress, one may not assume that the intermediate lobe alone plays an important role in the maintenance of the adrenal cortex, as the adrenals are similarly affected by hypophysectomy or pars distalis removal.


[Hormonal control of the adenyl cyclase activity of adipose cell membranes prepared from badger, rabbit, fox and rat adipose tissues].

1. We have shown differences in hormonal regulation of adenylate cyclase activity in fat cell ghosts prepared from rat, rabbit, fox and badger adipose tissue, under the influence of catecholamines, ACTH and insulin. a) In the rat, catecholamines induced a large stimulation (+315%) of adenylate cyclase. b) In the rabbit, ACTH was the most effective hormone. c) In the fox and the badger, only catecholamines could stimulate adenylate cyclase. d) In both rat and rabbit, insulin did not reduce spontaneous enzymatic activity. Moreover, the activation of adenylate cyclase by ACTH in the rabbit was not altered by insulin, while in the rat, this hormone slightly decreased epinephrine stimulation. 2. Hormonal regulation of adenylate cyclase correlated with the lipolytic response.


[Neuromechanisms of reciprocal interrelation between jaw-opening and jaw-closing muscles in the cat (author's transl)].

Neural controlling mechanisms between the digastric (jaw-opening) and masseter (jaw-closing) muscles were studied in the cat. High threshold afferent impulses from the anterior belly of the digastric muscle to masseteric montoneurons in the trigeminal motor nucleus induced an EPSP-IPSP sequence of potentials with long latency, and high threshold afferent impulses from the masseter muscle also exerted a similar effect on digastric motoneurons in the same nucleus innervating the anterior belly of the digastric muscle. These results suggest that reciprocal inhibition via Ia interneurons as observed between the flexor and extensor muscles in the spinal cord does not exist between the digastric and masseter muscles in the cat. However, the respective motoneurons innervating the masseter and digastric muscles receive inputs of early excitation-late inhibition via high threshold afferent nerve fibers from each antagonistic muscle. As such, since EPSPs preceding IPSPs are recognized, these high threshold afferent impulses may exert not only a reciprocal inhibitory effect, but also a synchronous excitatory or inhibitory effect on the antagonistic motoneurons.


Non functional malignant nesidioblastoma. About one case (author's transl).

The authors report a case of, non functional, malignant, delta-cell, nesidioblastoma and, this way, make a review of the literature. It was a cephalic tumor, which manifestations were jaundice and loss weight and which hypervascular character involved an abdominal bruit. The angiographic aspect settle them to established correlation between functional and non functional tumors: the formers characteristic "blush", agreeding in fact with the initial phase of the growth, increase in a monstruous "pseudoangiomatous" aspect in the laters.


Induction of DNA polymerase in polyoma virus-infected mouse cells requires transcription and translation.

The induction of DNA polymerase activity and of DNA synthesis in polyoma virus-infected mouse kidney cells is inhibited by actinomycin D and cycloheximide, even in cells that are already T-antigen positive. This indicates that the induction of the DNA-synthesizing apparatus requires transcription and translation. The increase in cellular DNA polymerase activity and in the DNA synthesizing capacity follows the appearance of T-antigen only after a characteristic lag period of several hours.


Avian leukosis viruses of different subgroups and types isolated after passage of Rous sarcoma virus-Rous-associated virus-0 in cells from different ring-necked pheasant embryos.

Avian leukosis viruses of subgroups A and F (RAV-A and RAV-F) arose at a low rate after passage of Rous sarcoma virus-Rous-associated virus-0, which is subgroup E, in cells from ring-necked pheasant embryos. In cells of two embryos, all of the viruses isolated after virus passage were RAV-F. However, in cells of a third embryo, both RAV-A and RAV-F were isolated. In addition, there sometimes were type-specific differences among the different isolates of RAV-A and RAV-F from the cells of single embryos. These results indicate that the RAV-A and RAV-F probably arose by recombination of viral and cellular genes, that different ring-necked pheasant embryo may have different endogenous avian leukosis virus-related nucleotide sequences, and that recombination at different sites in these endogenous sequences might give rise to type-specific differences among the RAV-A and RAV-F.


Action of ethyl and methyl methane sulfonates on DNA injection and genetic recombination in T7 bacteriophage.

After treatment with methyl or ethyl methane sulfonate, T7 amber mutants display a reduced capacity for recombination. Moreover, alkylation reduces recombination frequency involving markers on the right-hand side of the genetic map more than it reduces recombination frequency involving markers on the left-hand side. We interpret this to mean that alkylation can stop DNA injection at any point along the DNA molecule, and that T7 phage injects its DNA in a unique fashion starting from the end carrying the genes for early proteins.


Encephalomyocarditis virus RNA: variations in polyadenylic acid content and biological activity.

Encephalomyocarditis (EMC) viral RNA was isolated from purified virus grown in Ehrlich ascites tumor cells. The viral RNA was found to contain polyadenylic acid [poly(A)] regions that were very heterogeneous in length. Chromatography of the EMC viral RNA on oligo(dT)-cellulose columns separated the RNA into three distinct fractions (peaks 1 to 3). Approximately 20% of the EMC viral RNA appeared as peak 1, 40% as peak 2, and 40% as peak 3. The RNA in each fraction appeared to be intact as shown by co-sedimentation with 35S unfractionated EMC viral RNA in SDS-sucrose density gradients. Approximately 95 to 100% of peaks 1 and 3, and 60 to 70% of peak 2, reappeared at the same elution position after rechromatography on oligo(dT)-cellulose. The RNA in peak 1 contained poly(A) with an average length of 16 nucleotides, peak 2 contained poly(A) with an average of 26 nucleotides, and peak 3 contained an average of 74 nucleotides in its poly(A) region. The distribution in the three fractions, as well as the average length of the poly(A) moieties, was relatively unaffected by changes in the cell suspension medium used during infection. Finally, each of the three viral RNA fractions was assayed for biological activity using an infectious RNA assay on L-cell monolayers. Infectivity of the viral RNA was found to increase with poly(A) length, with peak 3 viral RNA being approximately 10 times more infectious than peak 1 viral RNA.


Size of murine RNA tumor virus-specific nuclear RNA molecules.

About 1% of the total RNA of cell lines producing murine leukemia virus is virus-specific RNA. About one-third of the virus-specific RNA is located within the nucleus. The size distribution of virus-specific RNA was determined before and after denaturation. Before denaturation, virus-specific RNA sequences sedimented as a heterogeneous population of RNA molecules, some of which sedimented very rapidly. After denaturation, most of the virus-specific RNA had a sedimentation coefficient of 35S or lower, but a small fraction of the nuclear virus-specific RNA sedimented more rapidly than 35S RNA even after denaturation.


Isolation and characterization of a heat-inducible simian virus 40 mutant.

We have isolated a new type of temperature-sensitive mutant of simian virus 40 (SV40) that is capable of productive infection in permissive cells but not of maintenance of viral DNA integration in transformed cells at the conditional temperature. Virus development is induced when cells transformed by this mutant are shifted to temperatures above 39 degrees C, but is not induced below this temperature. The plaque-purified, temperature-sensitive mutant virus confers heat inducibility to new host cells, indicating that the conditional function is a property of the viral genome. Unlike previously described temperature-sensitive SV40 mutants, in (ts)-1501 is capable of productive infection in permissive cells at the conditional temperature. The morphology, growth, and oncogenicity of in (ts)-1501-transformed cells at 37 degrees C are similar to those of cell lines transformed by wild-type SV40. HK10-c2(in(ts)-1501), a cloned cell line, transformed at 37 degrees C by the mutant virus, exhibits a transient increase in DNA synthesis before cell death at the conditional temperature. Many properties of in(ts)-1501 are analogous to those of the heat-inducible mutants of bacteriophages in which a heat-inactivated protein is responsible for the stable integration of the prophage in the bacterial chromosome.


Association of the polioviral RNA polymerase complex with phospholipid membranes.

Polioviral RNA polymerase complex, which consists of enzyme, template, and nascent RNA, is membrane bound in vivo. The solubilized RNA polymerase complex associated spontaneously in vitro with phospholipid bilayer membranes (liposomes) of defined composition. The degree of association at 37 degrees C was greater for those membranes that were more fluid, suggesting that the binding involves the interaction of the RNA polymerase complex with the hydrocarbon chains in the interior of the lipid bilayer. The polymerase activity was not enhanced by addition of the lipid; in fact, the addition of some of the longer-chain lipids resulted in up to a 40% inhibition of the polymerase activity. Spin-label electron paramagnetic resonance experiments, which measured the membrane fluidity, and kinetic experiments on the rate of incorporation of tritiated UTP into RNA by the polymerase were performed as a function of temperature. The results indicated that the activity of the polymerase was not affected by the physical state of the phospholipid membrane and that its active site was not intimately associated with the membrane. Analysis of both the viral and host polypeptides associated with the smooth membrane-bound polymerase indicated that X was the primary viral polypeptide present. In addition, host polypeptides of molecular weight 86,000, 62,000, 54,000, and 46,000 were also present. If the membrane was disrupted with detergent, polypeptide X was released from the polymerase activity, suggesting that X may play a role in binding the polymerase to the membrane. In an analogous manner, polypeptide X associated spontaneously with phospholipid membranes to a greater extent than the capsid polypeptides. Analysis of both the host and viral polypeptides associated with the viral RNA polymerase purified by precipitation in 2 M LiCl indicated that host polypeptides of molecular weight 106,000, 38,000, 33,000, and 14,000 were the major constituents, whereas relatively small amounts of the viral polypeptides were present. It was confirmed that of the viral polypeptides found, polypeptide 4 was present in the largest amount.


Analysis of in vitro transcription products of intracellular vesicular stomatitis virus RNA polymerase.

The intracellular transcriptase complex of vesicular stomatitis virus-infected L cells synthesized RNA complementary to the entire infectious virus genome at either 37 degrees C or 28 degrees C in vitro. Not all sequences were present at the same frequency, however; copies of that segment of the genome common to the LT defective particles were present at 20 to 100 times higher frequently than copies of the genome segment common to the ST defective particle. The less frequent region was transcribed somewhat more effectively at 28 degrees C than at 37 degrees C. The results suggest that transcriptional regulation rather than selective degradation is responsible for the differential accumulation of RNA.


Nucleotide sequence that binds primer for DNA synthesis to the avian sarcoma virus genome.

Initiation of transcription from the genome of avian sarcoma virus by RNA-directed DNA polymerase in vitro requires tRNAtrp as a primer. The tRNA is bound to the viral genome by a sequence of 16 contiguous nucleotides (U-C-A-C-G-U-C-G-G-G-G-U-C-A-C-Cp), beginning with the penultimate base at the 3' terminus of the primer and extending through the acceptor stem into loop IV of the tRNA. Consequently, the native conformation of the tRNA must be disrupted by the binding of primer to the viral genome. The binding sequence does not include two adjacent residues of pseudouridine in loop IV, which distinguish the primer from many other tRNAs, and the 3' terminal adenosine of primer may also be excluded from base pairing with the viral genome.


Conditional lethal mutants of adenovirus type 2-simian virus 40 hybrids. II. Ad2+ND1 host-range mutants that synthesize fragments of the Ad2+ND1 30K protein.

Adenovirus type 2 (Ad2) grows 1,000 times less well in monkey cells than in human cells. This defect can be overcome, not only upon co-infection of cells with simian virus 40 (SV40), but also when the relevant part of the SV40 genome is integrated into the adenovirus genome to form an adenovirus-SV40 hybrid virus. We have used the nondefective Ad2-SV40 hybrid virus Ad2+ND1, which contains an insertion of 17% of the SV40 genome, to isolate host-range mutants which are defective in growth on monkey cells although they grow normally on human cells. Like Ad2, these mutants are defective in the synthesis of late proteins in monkey cells. A 30,000-molecular-weight protein (30K), unique to Ad2+ND1-infected cells, can be synthesized in vitro, using Ad2+ND1 mRNA that contains SV40 sequences. 30K is not seen in cells infected with those host-range mutants that are most defective in growth on monkey cells, and translation in vitro of SV40-specific mRNA from these cells produces new unique polypeptides, instead of 30K. Genetic and biochemical analyses indicate that these mutants carry point mutations rather than deletions.


Translation of individual host mRNA's in MPC-11 cells is differentially suppressed after infection by vesicular stomatitis virus.

Infection of MPC-11 mouse plasmacytoma cells by vesicular stomatitis virus results in 30 to 35% reduction in [35S]methionine incorporation into total proteins within 30 min postinfection. By 6 h postinfection, total protein synthesis is reduced by 80 to 90%. However, even by 30 min postinfection, a differential suppression of the synthesis of individual host protein is observed. The synthesis of the immunoglobin G (IgG) heavy chain (H), and, in particular, the synthesis of IgG light chain (L), is considerably more resistant to vesicular stomatitis virus-induced inhibition than is the synthesis of the non-IgG proteins as a whole; e.g., when the synthesis of non-IgG proteins was reduced by 41%, the synthesis of the H and L chains was reduced by 28 and 7%, respectively. Furthermore, these alterations in the relative synthesis of the L chain, H chain, and non-IgG are comparable to the alterations previously observed in uninfected MPC-11 cells when the overall rate of polypeptide chain initiation was selectively reduced (D.L. Nuss and G. Koch, 1976). These results are discussed in terms of the strategy of virus-directed suppression of host mRNA translation.


Human cytomegalovirus: glycoproteins associated with virions and dense bodies.

The glycoproteins associated with the membranes of cytomegalovirions and dense bodies were characterized by their relative mobility, percentage of glucosamine incorporation, and molecular weight. Eight glycopolypeptides were repeatedly detectable. Three glycopolypeptides of higher molecular weight with low levels of glucosamine incorporation were occasionally detectable. These latter glycopolypeptides may be precursors or aggregates of the glycopolypeptides with lower molecular weights. The glycoproteins associated with the membranes were on the surface, as determined by iodination with 125I of virions and dense bodies partially purified in gradients of D-sorbitol. Velocity centrifugation in linear gradients of D-sorbitol was used to obtain concentrated and partially purified preparations of infectious cytomegalovirus. Viral infectivity and the membranes of cytomegalovirions and dense bodies were stable in gradients of sorbitol, but cellular contaminants were not completely removed. Additional centrifugation in CsCl separated both cellular contaminants and viral nucleocapsids from virions and dense bodies. Many dense bodies, which are considered to be aberrant forms of cytomegalovirus, had the same size, sedimentation properties, and density as virions. Consequently, they were not separable from virions by various centrifugation techniques. Electron microscopy demonstrated that purified virions and dense bodies were qualitatively free of extraneous material and that each dense body was bounded by a membrane, as evidenced by its double-tract appearance. Antisera to a preparation of purified virions and dense bodies, or to their glycoproteins, contained antibodies that neutralized viral infectivity and reacted with antigens in cells infected with cytomegalovirus. However, these same antisera did not contain antibodies that reacted with uninfected cells. The glycoproteins associated with the membranes of cytomegalovirions and dense bodies are considered to be specified by the cytomegalovirus genome.


RNA polymerase activity in purified virions of avian reticuloendotheliosis viruses.

An RNA polymerase activity that synthesizes a U-rich RNA hydrogen bonded to a large viral RNA molecule was found in the cores of virions of avian reticuloendotheliosis viruses (REV). The RNA polymerase activity was separable from the DNA polymerase activity of REV virions. The 5'-terminus of the newly synthesized RNA was A. In addition, a tRNA nucleotidyl transferase activity, which added -CpCpA ends to tRNA, appears to be present in the REV virions.


Early events in polyoma virus infection: attachment, penetration, and nuclear entry.

The plaque-assay technique was used as a tool to determine the optimal conditions for adsorption of polyoma virions to host cells. Using these optimal conditions of adsorption, an electron microscopy study of the early events of infection was performed. By electron microscopy and autoradiography, it was demonstrated that both the viral coat proteins and DNA arrive simultaneously in the nucleus as early as 15 min postinfection. When horseradish peroxidase-labeled virions, pseudovirions, and capsids were used to infect cells, only the particles with nucleic acid or a factor(s) associated with the nucleic acid, i.e., histones, appeared to enter the nucleus. Moreover, when virions were used to infect either permissive or nonpermissive cells, identical early events of viral infection, i.e., adsorption, penetration, and nuclear transport, were observed, suggesting that these early events of infection are a property of the virion and not the host cell.


Comparison of JC and BK human papovaviruses with simian virus 40: DNA homology studies.

Studies were performed to ascertain the relationship of human papovavirus JC to BK virus and to simian virus 40 (SV40) by further restriction endonuclease analysis and by DNA-DNA competition hybridization on membrane filters. Form I DNA extracted from two new isolates from cases of progressive multifocal leukoencephalopathy of human papovaviruses that were JC-like in their antigenic properties were found to yield restriction endonuclease fragmentation patterns similar to those of prototypic JC virus DNA and different from those of BK or SV40. Form I DNA preparations of JC and BK viruses were found to be related to each other and to SV40 DNA to a similar extent, with JC and BK virus DNAs containing sequences homologous to both early and late regions of the SV40 genome. The relatedness in each comparison was less than 50%, and heterologous hybrids between either JC or BK and SV40 DNAs were found to be less stable than homologous SV40-SV40 hybrids in high concentrations of formamide, suggesting substantial mismatch within homologous regions, to the extent of 15 to 30%. The new JC-like isolates were also studied in competition hybridization reactions with SV40 DNA and yielded results similar to those obtained with JC virus.


Differential response of type C and intracisternal type A particle markers in cells treated with iododeoxyuridine and dexamethasone.

Mouse neuroblastoma cells containing intracisternal type A particles were treated with iododeoxyuridine and dexamethasone to induce the release of type C oncornavirus particles. For 5 days after treatment, antigenic markers and DNA polymerase activities specific to particles of each of the two types were assayed in the cells and in pellets obtained by high-speed centrifugation of the culture fluid. There was a marked release of C-particle antigen (p30) and DNA polymerase activity in extracellular particulate form, reaching a maximum on day 3 after treatment and falling thereafter. In contrast, no extracellular A-particle antigen was detected, and A-particle-specific DNA polymerase activity in the medium pellets did not increase from the original very low level. Electron microscopy confirmed the presence of free type C virus particles, but not intracisternal type A particles, in the culture fluid. Although intracellular levels of C-particle antigen rose 20- to 30-fold per milligram of cell protein, intracellular A-particle antigen and DNA polymerase activity did not vary more than two-fold. The relative rate of A-particle synthesis in the treated cells, as judged by incorporation of radioactive amino acids into the major structural protein (P73), was also unchanged over the period of observation. Thus, the induction of type C virus particle formation in cultured neuroblastoma cells had no detectable effect on the quantity, synthesis rate, or location of intracisternal type A particles.


DNA-binding proteins induced by herpes simplex virus type 2 in HEp-2 cells.

Affinity chromatography on single-stranded and double-stranded DNA-cellulose indicates that 12 proteins previously identified from herpes simplex virus type 2-infected cells, ranging in molecular weight from 28 X 10(3) to 186 X 10(3), bind to DNA-cellulose. The DNA-binding proteins found in infected cells differed in relative binding strengths for denatured DNA-cellulose. The virus specificity of these DNA-binding proteins was further studied by comparison with DNA-binding proteins isolated from mock-infected cells, and by immunoprecipitation of infected-cell DNA-binding proteins with antisera specific for viral antigens. The promise this technique holds for the purification and study of polypeptides involved in virus DNA replication, recombination, or repair is discussed.


Polyadenylic acid in the genomic RNA of mengovirus.

The polyadenylic acid contained in 35S mengovirus RNA produced in infected BHK-21 cells contained approximately 94% AMP and was estimated to contain an average of 50 to 55 nucleotides. The polyadenylic acid is placed at the 3'-end of the genomic RNA based on the presence of significant levels of [3H]adenosine in complete alkali or RNase T2 digests of polyadenylic acid from [3H]adenosine-labeled 35S viral RNA.


Interaction of liposomes with subviral particles of poliovirus type 2 and rhinovirus type 2.

Subviral particles ("A particles") were produced from rhinovirus type 2 by treatment with acid and from poliovirus type 2 by incubation at 37 degrees C in a low-ionic-strength buffer. A particles, but not virions, adsorbed to liposomes. It is proposed that these reactions may provide an in vitro model for two early steps of infection.


Radioisotopic labeling of human papovavirus (BK) by iodination and reductive alkylation.

Purified virions of the GS strain of the BK group of human papovaviruses were labeled with 125I using chloramine T or lactoperoxidase or with tritium using sodium borohydride. All viral polypeptides were labeled. Tryptic digests of iodinated VP1 were analyzed.


Lysine tRNA's associated with avian myeloblastosis virus 70S RNA.

The lysine tRNA released from the 70S RNA of avian myeloblastosis virus was separated by reversed-phase chromatography. All of the AAG-coding lysine tRNA's were present in the 70S-associated fraction; however, the AAA-coding lysine tRNA could not be detected. Chromatography of the lysine tRNA released at various temperatures did not show any preferential release of one AAG-coding species over another.


Hepatic adenomata with type 1 glycogen storage disease.

In older patients with glucose-6-phosphatase deficiency adenomatous nodules develop within the liver parenchyma. Investigation of eight such patients, age 3 to 28 years, using radioisotopic scans, has demonstrated areas of depressed isotope uptake in the liver in all except the one preteenaged child. Three patients were further studied with hepatic angiograms and liver biopsy specimens. A diffuse nodularity, more widespread than apparent on isotopic scans, was demonstrated on angiograms. Although initial histologic study in each case showed adenomatous tissue without evidence of neoplasm, the development of hepatocellular carcinoma in one of our patients and in others from the literature suggests that the nodules may be premalignant.


Positive differential heterophile antibody test. Persistence in a symptomatic patient.

A patient who had documented mononucleosis in June 1971 has been persistently symptomatic since that time, with easy fatigability and weight loss. He has also had a positive differential heterophile antibody test for mononucleosis for 14 months. Although no evidence for active Estein-Barr virus (EBV) infection was discovered, the possibility of a chronic EBV infection is discussed.


Hepatic abscess. Diagnosis in the adult by total body opacification.

Two adults had the hypovascular nature of their hepatic abscesses ascertained by total body opacification. In one, total body opacification identified the hypovascular nature of the lesion, facilitating its differentiation from vascular hepatoma. In the other, the clinical diagnosis was acute cholecystitis with empyema of the gallbladder, and the liver was normal to inspection and palpation at celiotomy. Total body opacification may be helpful in the examination of abdominal masses.


Clostridium perfringens exotoxins. III. Binding of theta-toxin to erythrocyte membrane.

When Clostridium perfringens theta-toxin was incubated with sheep erythrocytes the toxin activity disappeared before lysis, the fact of which suggests fixation of the toxin to erythrocyte membranes. 2. Theta-Toxin lost its activity by binding to cell membranes, and the membrane constituted inhibitor of theta-hemolysis was neither a protein, a carbohydrate nor a phosphatide, but was cholesterol. From these results this report proposes that the theta-toxin binding site of erythrocytes should be cholesterol.


Vertical transmission of type I and type 2 Herpes simplex virus infection in hamsters.

The relation of type 2 herpes simplex virus infection during the several stages of embryogenesis of hamster was examined using histopathological and immunofluorescent techniques. The comparative investigation between type 2 and type 1 was done. Both types of herpes simplex virus caused placental lesions and vertical transmission in fetuses when the virus was inoculated intravenously into pregnant hamsters at the middle and late stage of gestation.


Temporal analysis of cellular cytotoxicity and humoral factors during progession and regression of Rous sarcomas in Japanese quails.

Temporal appearance of cellular cytotoxicity and humoral activities including blocking and arming activities during the entire course of Rous sarcoma development in Japanese quails was examined by microcytotoxicity assay with comparison of animals bearing regressing tumors induced by a moderate dose of virus (regressors) and animals bearing growing tumors induced by a large dose of virus (progressors). Cellular cytotoxicity of the spleen cells in regressors was detected in a biphasic pattern; the first phase being observed as early as 3-5 days post inoculation (p.i.), followed by an eclipse period between 7-10 days p.i. which was the time of active tumor growth, and the second phase occurring after 12 days p.i. when the tumor had attained the maximum size. In progressors, only the first phase was observed. Instead, a stimulatory effect of the spleen cells on growth of target cells was noticed. Arming activity which confers cytotoxic activity on the normal spleen cells was demonstrated in the sera of regressors in the similar biphasic pattern as the cellular cytotoxicity; the early activity being present at 3 days p.i., and the late one after 19 days p.i. The former was detected by pre-incubation of serum with effector cells in microcytotoxicity assay and the latter by pre-incubation with target cells. In progressors, only the early arming activity which reacts with effector cells was demonstrated. Blocking activity which abrogates cellular cytotoxicity was demonstrated in both regressors and progressors but in different patterns of appearance, that is, blocking activity in regressors was only transiently demonstrated only by pre-incubation with effector cells at the time of maximum tumor growth, while the activity in progressors seemed to persist after the tumor reached the maximum size. Since the earlier activity was found to be effective at effector cell level, and the later one at both effector and target cell levels, participation of blocking factors of different types in progressors was also suggested.


[Gene frequencies of red cell adenylate kinase and adenosine deaminase in the population of Northern Bavaria (Germany) (author's transl)].

Within a population sample of 2,000 blood donors in Northern Bavaria the AK- and ADA-polymorphisms have been investigated. The calculated gene frequencies are: AK1=0,9665; AK2=0,333; AK3=0,0003; AKA1=0,9453; ADA2=0,0548.


Redistribution of immunoglobulin receptors on human neutrophils and its relationship to the release of lysosomal enzymes;.

Release of the lysosomal enzyme beta-glucuronidase from human neutrophils was induced by IgG or its Fc fragment, aggregated by immune precipitation or by coating on latex particles. Such release was inhibited when the cells were preincubated with free IgG or Fc fragments; F(ab')2 fragments were ineffective in both inducing and inhibiting beta-glucuronidase release. Neutrophils incubated with IgG or Fc fragments, when challenged with anti-IgG antibody, released lysosomal enzymes without the release of the cytoplasmic marker lactic dehydrogenase; These studies indicate that human neutrophils have surface receptors for the Fc portion of IgG. Neutrophils treated with IgG or its Fc fragment and subsequently with fluorescein- or ferritin-labeled anti-IgG showed binding of Fc or IgG to the cell membrane. Under suitable conditions, polar capping of labeled antibody was seen by fluorescence or electron microscopy. These studies suggest that the immunoglobulin receptors on neutrophils are redistributed when they are cross-linked with antibody. Fluidity of the membrane receptors appeared to be time and temperature dependent. Compounds such as 2-deoxyglucose, colchicine, and cyclic AMP, which inhibit the release of lysosomal enzymes, also inhibited the redistribution of the surface receptors. Cytochalasin B, an agent which increases the release, was found to increase the receptor redistribution; The relationship between the release of lysosomal enzymes and receptor mobility is discussed;


An ultrastructural study of central nervous system disease produced by wild-type and temperature-sensitive mutants of vesicular stomatitis virus.

Outbred Swiss mice 3 to 4 weeks of age were injected intracerebrally with wild-type vesicular stomatitis virus or its temperature-sensitive (ts) mutants ts 11, ts 22, ts 31, and ts 41. Brain and spinal cord were then studied for pathologic changes by electron microscopy. All mice infected with wild-type vesicular stomatitis virus died within 2 days of inoculation. Diffuse ependymal alterations often culminating in necrosis in brain and especially spinal cord and rare foci of necrosis and mononuclear cell infiltration in the injected hemisphere were the main pathologic changes in these animals. In contrast, mice infected intracerebrally with ts 22 and ts 31 showed their first clinical signs, consisting of hind limb paralysis, on day 4 and did not die until day 7 or 8 after inoculation. Ependymal alterations were of less severe degree in these animals, whereas the most striking changes were those of status spongiosus limited to the gray matter of the spinal cord. Such status spongiosus was mainly due to ballooning of dendrites and astrocytic processes, although myelin and neurons were also occasionally involved. Mice infected with ts 11 and ts 41, on the other hand, remained clinically well and failed to show significant pathologic features at 4 and 8 days after intracerebral inoculation. This study would indicate that some ts mutants of vesicular stomatitis virus are capable of altering the fulminating disease produced by the parent virus and of producing strikingly different pathologic changes.


Ultrastructural study of contraction of pulmonary vascular smooth muscle cells.

The pulmonary vessels of rats treated with fulvine were studied electron microscopically for morphologic signs of contraction of smooth muscle cells. Except for a number of indirect indications of vasoconstriction such as medial smooth muscle cell hypertrophy and excessive crenation of elastic laminae, conspicuous smooth muscle cell excrescences were observed and were interpreted to be a direct result of contraction. The close relationship between contraction and smooth muscle cell excrescences was confirmed by their simultaneous occurrence within 1 minute after administration of histamine to isolated perfused guinea pig lungs. The images of pulmonary vessels of rats with prolonged survival times after fulvine administration suggested a gradual increase in the size of the excrescences with a simultaneous degeneration of their cytoplasmic content, and in some cases their eventual detachment from the main cell body. The latter changes were possibly associated with the widespread vasculitis that often occurred at longer intervals after fulvine application. The smooth muscle cell excrescences in pulmonary veins were generally much more prominent than those in pulmonary arteries. This difference was probably caused by the more rigid structure of the arterial wall whcih prevented the formation of large excrescences. The mechanism of the formation of smooth muscle cell excrescences, their possible general validity as markers of vascular smooth muscle cell contraction, and the implications for the mechanism of action of fulvine are briefly discussed.


An ultrastructural study of the glomerular slit diaphragm in New Zealand black/white mice.

Glomerular epithelial slit alterations and their relation to proteinuria have not been studied in detail in New Zealand Black/White (NZB/W) mice. The kidneys of proteinuric and nonproteinuric female NZB/W mice and normal Swiss albino mice were perfusion-fixed with tannic acid-glutaraldehyde and studied by light and electron microscopy. Semiquantitative studies were performed on full montages of glomeruli enlarged 10,000 times. Fine structural alterations of the epithelial slits, with emphasis on the slit diaphragm, were studied on semiserial thin sections. Proteinuric NZB/W mice with features of membraneous nephropathy exhibited: (1) wedging of electron-dense deposits below the slit diaphragm, (2) enlargement and distortion of interpedicel spaces, (3) displacement, folding, and stacking of slit diaphragms, (4) formation of occluding junctional complexes in residual slits, and (5) variable loss of foot processes. Similar alterations were not observed in controls or nonproteinuric NZB/W mice, including animals having complexes inglomerular mesangia but not in epithlialslits. These studies show that in NZB/W mice, abnormal protein excretion is associated with structural modification of the slit pore and suggest a role for such a component in the process of protein ex


Assessment of skeletal aging using the 3H-proline topographic labeling method.

The present report represents an extension of initial 3H-proline autoradiographic studies designed to provide, at both the cytologic and histologic levels, an indelible topographic record of skeletal events in aging mice. 3H-proline was administered in multiple time-spaced doses and the animals were serially killed between 6 and 14 days later. Whole femora sectioned in longitudinal and three cross-sectional planes were prepared for autoradiography and subsequent grain count and micrometric analysis of the silver topographic bands which form over mineralizing surfaces. These bands, coincidental with the cellular uptake and turnover of radiotracer, allowed for the assessment of cellular rates of labeled matrical precursor production and deposition from which the processes of bone growth and remodeling and the effects of aging on the skeletal system can be evaluated. In addition, composite tracings of topographic data derived from multiple sections prepared at different geometric planes allowed for three-dimensional mapping of skeletal events. The results showed that the matrical rates of bone production in micrometers per day varied considerably from one surface to the next and decreased significantly with increasing age. The rates at posterior periosteal surfaces were twice those observed at the anterior periosteal surfaces of 5-week-old mice. By 26 weeks the rates decreased to 1/8 times, respectively. Between 52 and 104 weeks of age, no significant matrix production occurred at the periosteal surfaces. Endosteal activity was greater than twice the posterior periosteal rate at 5 weeks, 1/4 times at 26 weeks, and 1/8 times at 104 weeks of age. It was concluded that endosteal matrix production continued throughout life via active remodeling processes, but periosteal activity was turned-off by 52 weeks. Furthermore, the study unexpectedly showed that in old animals, subendosteal bone formation exhibiting hematopoietic activity is activated at 52 weeks in focal areas...


Crystals, paracrystals, and rigid tubules in multiple sclerotic brain and spinal fluid.

Brain and spinal fluid were obtained post mortem from an individual with multiple sclerosis who died following carbon monoxide inhalation. Electron microscopic examination of the plaque region revealed a crystalline structure (repeat unit 380 to 410 A) within rough endoplasmic reticulum of astrocytes, a paracrystal (160 to 220 A) within the cytoplasm of astrocytes, and rigid tubules (outer diameter 320 to 450 A) within a cell in the lumen of a capillary. The paracrystal was enclosed by the glial fibrils. A second crystalline material was observed within cells in the 15,000 X g pellet from spinal fluid. The ultrastructural pattern of this second crystal was generated by three sets of parallel lines; a given set was intersected by the other two sets. The crystal had only two different repeat units (410 to 450 A; 310 to 350 A) since the smaller dimension was common for two sets of lines. The two crystals were large; the one in astrocytes had a length of 52,000 A and that in spinal fluid sediment was 62,000 A. These structures were observed in the one multiple sclerotic brain obtained at autopsy but were not detected in multiple sclerotic brains that had been frozen for extended periods. Plaque regions and grossly normal white matter were homogenized and the 1000 X g supernatant of this material was fractionated in a continuous CsCl-sucrose density gradient. Long tubules (320 to 500 A diameter) were observed in the 1.24 to 1.26 density region. The major proteins of the 1.24 to 1.26 fraction had molecular weights clustered at 40,000 to 45,000, 88,000 to 130,000, and 300,000.


The malignancy of bronchial adenoma.

The literature on bronchial adenoma has been reviewed and controversy found to exist about certain aspects of these tumors, particularly their malignancy. A retrospective study of 79 cases managed in Edinburgh since 1946 is presented. Seventy-one were of the carcinoid type, 7 were adenoid cystic carcinomas, and there was a single example of mucoepidermoid carcinoma. All types show a definite malignant potential, greatest in the rare mucoepidermoid tumor and least in the common carcinoid variety. Adenoid cystic carcinoma occupied an intermediate position in frequency and malignancy.


Immune reactivity in primary carcinoma of the lung and its relation to prognosis.

Detailed studies of immune reactivity were performed in 154 patients with primary lung cancer, 20 patients with benign thoracic lesions, and 109 healthy persons. Reactions to the 2,4-dinitrochlorobenzene (DNCB) skin test were postive in 73 per cent of patients with lung cancer and all (100 per cent) of the patients with benign disease (p less than 0.05). The incidence of DNCB reactions was 78 per cent for Stage I and II cancers (37 patinets), 73 per cent for resectable Stage III cancer (22 patients), and 66 per cent in patients with unresectable or inoperable Stage III cancer. DNCB reactivity showed a relationship to primary histology. The incidence of DNCB positive reactions was 80 per cent in patients with epidermold carcinoma versus 57 per cent in patients with adenocarcinoma, 64 per cent in patients with oat cell cancer, and 80 per cent in patients with terminal bronchiolar carcinoma. In vitro immune studeis correlated best with stage of disease. These included the absolute lymphocyte count and absolute T cell count and lymphoxyte stimulation witalen A (Com A). These values were in the normal range in patients with Stage I cancer but were significantly depressed in patients with Stage III cancer. Svrvival curves were plotted in patients with Stage III disease according to the responses to three immune parameters: DNCB, absolute lymphocyte count, and PHS stimulation. Although patients with normal reactions generally had better survival rates, PHA responses showed the most significant correlation to survival. These tests support the usefulness of immune testing as an additional parameter of assessing biological risk in patients with primary lung cancer.


Results of treatment of primary carcinoma of the lung. Analysis of 3,000 cases.

Three thousand patients with primary carcinoma of the lung entered in the Armed Forces Central Medical Registry are reported. Forty-one per cent had squamous cell, 28.5 per cent adenocarcinoma, 25.2 per cent small cell/undifferentiated, and 4.9 per cent miscellaneous cell types. When first seen, 71.1 per cent had no organ metastases and 50.6 per cent no lymph node metastases. Over-all survival rate was 18.2 per cent at 5 years and 14.5 per cent at 10 years. Survival following definitive resection, palliative resection, definitive radiation, palliative radiation, and chemotherapy was determined both in the presence of mediastinal nodal involvement and in the absence of mediatinal nodal involvement. Where resection for cure could be carried out, 5 year survival rates of 48.8 per cent were possible. The factors affecting this improved outlook in our military population are discussed and, in general, appear to be related to a ready accessibility of medical care and the necessity, because of global commitments, of establishing an early diagnosis. Cell type ecerted some influence on survival, but the major determinant appeared to be the absence of involved nodes at the time of the operation.


Ultrastructural cytochemistry of platelets and megakaryocytes in the carcinoid syndrome.

Platelets and megakaryocytes from 11 patients with the carcinoid syndrome have been studied by transmission electron microscopy. Cells fixed in phosphate-buffered glutaraldehyde are oval to discoid, with pseudopods, a dilated open-channel system, and a prominent dense tubular system as defined by peroxidase activity and alkaline bismuth stain. Atypical with hexagonal lattices and treaded substructures and large (diameter greater than 0.5 mum), phosphatase-positive, debris-containing vacuoles are four times more numerous than in normal platelets. Incubation of platelets in a 0.05% suspension of latex results in particle incorporation into phagosomes and the debris-containing vacuoles. Molybdate-dichromate stain reveals two classes of dense bodies, one of which (with a reticular core) is 20 times more numerous than in normal platelets. Bone marrow megakaryocytes lack both dense bodies and debris vacuoles analogous to those found in circulating platelets. These results suggest autophagy or endocytosis abnormalities and provide evidence for multiple types of dense bodies in carcinoid platelets.


[Etiological aspects regarding the development of laryngeal papillomas (author's transl)].

Papilloma of the larynx are virus-induced, benign neoplasmas. The virus belongs to the Papovavirus group and causes skin-warts, condylomata acuminata and papilloma of the mucous membranes. The close relation of these diseases is shown by a review of dermatologic and otologic literature and compared with own results.


Treatment of senile dementia and cerebellar disorders with phthalazinol. Cyclic AMP-increasing agent, phthalazinol, in therapeutic trials in hitherto incurable morbid conditions (I).

Three patients suffering from presenile dementia and two patients suffering from senile dementia were treated with phthalazinol. A limited but definite improvement in attentiveness, vocabulary reception and psychologic attainment and also a limited but definite improvement in range of motion, gait and endurance were noted shortly after the treatment with phthalazinol and the progression of their dementia seemed to be temporarily retarded by phthalazinol. Seven patients suffering from parenchymatous cerebellar degeneration (late cortico-cerebellar atrophy) and four patients suffering from olivopontocerebellar atrophy were treated with phthalazinol. A relatively rapid, limited and sustained improvement of cerebellar functions, including those of speaking, writing and walking, was noted in almost all patients. Also, rigidity, akinetic tendency and abnormal posture seen in olivopontocerebellar atrophy fairly well responded to the treatment. Dysphagia, disturbance of micturition, and hypersecretion of saliva have also rapidly and completely disappeared in all cases. The cases with a long history responded relatively poorly, but the cases with a relatively short history responded quite strikingly to the therapy. No side effects were noted.


Evaluation of an automated procedure for primary screening of hyperlipoproteinemias.

An automated procedure for the determination of beta-plus-pre-beta-lipoprotein cholesterol is described and evaluated for mass screening of hyperlipoproteinemias. It is shown that this method fulfills the requirements for an ideal method for primary screening: it can be used in non-fasting samples, is capable of detecting hyperlipoproteinemias characterized by both cholesterol and/or triglyceride abnormalities, is fast, accurate and extremely inexpensive. Evidence is shown that the method is extremely useful for primary screening and, when tested in a coronary screening detection program, gave only 4% false positive and 3.4% false negative results.


Therapeutic failure in familial type II hyperlipoproteinemia.

The extended use of diet and cholestyramine therapy in familial type II hyperlipoproteinemia was examined in patients who previously participated in a short-term, double-blind trial. A striking secondary failure in therapeutic response during 4 yr of use of this therapy was noted with plasma cholesterol rising an average of 15%. A 3 mo, out-patient, follow-up study designed to reinforce patient motivation and dietary and drug adherence resulted in a prompt but partial reversal of this therapeutic deterioration in 16 patients. Additional inpatient studies confirmed that patient noncompliance with the dietary regimen was the major factor responsible for the secondary failure. Cholestyramine together with a low cholesterol diet can be an effective agent in familial type II hyperlipoproteinemia, given a comprehensive program of out-patient follow-up with continued emphasis on dietary principles and drug adherence.


Low density lipoprotein metabolism in a family of familial hypercholesterolemic patients.

A low-density lipoprotein turnover study was performed on six heteroxygous familial hypercholesterolemic subjects and five control subjects. The diagnosis of the condition was clear-cut on biochemical, clinical, and genetic grounds. Kinetic analysis of the plasma decay curves gave the following mean values: (1) The plasma concentration of low density lipoprotein apoprotein (apoLDL) in the affected subjects was 247 mg/dl, a 2.5-fold increase over control values (98 mg/dl). (2) The calculated fractional catabolic rate of the intravascular apoLDL pool in the dyslipo-proteniemics was reduced with respect to control data (16.4%/day versus 31.2%/day), and the half-life of the apolipoprotein was correspondingly increased (6.07 days versus 3.63 days). (3) The absolute catabolic rate of the apoLDL was 15.6 mg/kg/day in contrast to the lower value of 11.6 mg/kg/day in the control group. (4) A strong negative correlation was observed between the plasma apoLDL concentration and the fractional catabolic rate (r = 0.96). We conclude from our data that increased apoLDL synthesis coupled with a defective or saturated catabolic mechanism is implicated in the pathogenesis of familial hypercholesterolemia.


Declining mortality and improving survival from Wilms' tumor.

Mortality from kidney cancer in the United States declined dramatically over the period 1940-1969. This trend was nearly proportional to the increases in survival for 644 U.S. children reported to the End Results Group, National Cancer Institute. The improvements in survival occurred for all stages of disease and appeared to result from therapeutic advances rather than earlier detection. Total incidence trends showed no consistent change, but a shift toward female preponderance resembled that seen in the mortality statistics and suggests sex differences in etiology.


Acute leukemia in Burkitt lymphoma.

Burkitt lymphoma uncommonly presents as a acute leukemia. We describe the clinical course and findings of a 14-year-old female with Burkitt lymphoma who presented with acute leukemia. splenomegaly and an abdominal mass. She responded initially to prednisone alone and later achieved full remission with combination chemotherapy. Established morphologic criteria and clinical course were consistent with the diagnosis of Burkitt lymphoma (1), although B-cell determinants were not present on her tumor cells.


Simultaneous determinations of Fe, Cu, Zn, and Br concentrations in human tissue sections.

Proton-induced x-ray fluorescence has been used to determine the concentration of trace elements in human prostate and liver tissue samples. Ca, Fe, Cu, and Zn were observed in all specimens. Small amounts of Br were detected in most of the prostates, and in nearly half of the livers. The data indicate that the Fe content decreases in malignant liver tissues and the quantities of Cu and Zn are also decreased, but to a lesser extent.


Effect of different carbon sources on the biosynthesis of polyunsaturated fatty acids of alpha-linolenic acid family in culture of minimal deviation hepatoma 7288 C cells.

The effect of three different carbon sources on the biosynthesis of polyunsaturated fatty acids of the alpha-linolenic acid series was investigated in hepatoma tissue culture (HTC) cells. Alpha linolenic acid was converted to higher homologs by a desaturating route that synthetized mainly 18:4 (delta6, 9, 12, 15), 20:4 (delta8, 11, 14, 17) and 20:5 (delta5, 8, 11, 14, 17) and an elongating route that produced 20:3 (delta11, 14, 17) and 20:4 (delta5, 11, 14, 17) acids. "Fasting" decreased both biosynthetic routes whereas glucose reactivated only the elongating pathway. Lactabumin hydrolysate enhanced significantly only the desaturating route whereas glycerol was inactive. Glucose and aminoacids increased similarly the incorporation of labeled alpha linolenic acid in the cells. The results are independent of hormonal effects.


Interaction of aminoacyl-tRNA synthetases and tRNA: positive and negative cooperativity of their active centres.

The influence of tRNA on the kinetics of PP-ATP exchange and aminoacyl-tRNA formation catalysed by leucyl-, phenylalanyl-, and tryptophanyl-tRNA synthetases has been investigated. These enzymes were chosen because they belong to three main classes of quaternary structure alpha1, alpha2beta2 and alpha2, respectively. The present paper shows that the investigated synthetases manifest kinetic cooperativity of the active centres which is negative in the case of AAA formation and positive in the case of leucyl- and tryptophanyl-tRNA synthesis. The obtained data were interpreted with the aid of the trigger model of the enzyme.


[Reaction capabilities and structure of poly(rG) and poly(rG)-poly(rC) in solution by the method of the kinetics of hydrogen ion exchange].

Data on the kinetics of 1H greater than 3H exchange between water and C(8)H groups of guanylic residues in the poly(rG) and poly poly(rG)-poly(rC) are presented. Furthermore, optical properties (CD spectra and hyperchromism) of neutral solutions of these polymers from 20 to 100 degrees C are described. It is shown that the exchange in poly(rG) within the temperature range from 20 to 80 degrees C proceeds faster than in rGMP. Within the temperature range from 20 to 40 degrees C such an acceleration of the exchange is observed also in poly(rG)-poly(rC). According to the ylide mechanism of the exchange reaction the observed accleration of the exchanged in in C(8)H groups of guanylic residues is considered as a consequence of an increase of the positive charge at N(7) atoms. This effect is due to formation of additional hydrogen bonds in which N(7) atoms take part. The exchange in poly(rG)-poly(rG) at temperatures hihger than 75 degrees C, when these additional hydrogen bonds are absent, proceeds more slowly than in rGMP. Such picture is usual in other previously studied polynucleotides whose structure in solution is stabilized only by Watson - Crick hydrogen bonds and stacking interactions. The data obtained support a Guschelbauer's model of the four-stranded stranded poly(rG). They also indicate the posibility of associates formation in poly(rG)-poly(rC) solutions at temperature lower than 40 degrees C being stabilized by hydrogen bonds in which N(7) atoms of guanylic residues take part.


[Determination of the distance between spin labels and the paramagnetic center in spin-labeled proteins according to parameters of the saturation curves of the label EPR spectra at 77 degrees K].

A new method of estimation of the distance between spins of the spin-label and paramagnetic center is suggested. Method is based on the quantitative analys of saturation curve of spin-label EPR spectra at 77 degrees K. New approaches have been tested using haemoglobin labeled on SH-groups with various iminoxyl radicals. Values of the distances between labels and haem estimated from the saturation curve parameters and by current methods and values of distances estimated from X-ray data are in good agreement. In the case of rapid spin relaxation of he paramagnetic center, the new method allows one to determine a farther distance. Results of the present work make it possible to investigat by the spin-label technique the structure of haem-containing propeins and the structure of other proteins with known spin relaxation time of paramagnetic center.


Radiation enhanced reactivation of herpes simplex virus: effect of caffeine.

Ultaviolet enhanced (Weigle) reactivation of UV-irradiated herpes simplex virus in UV-irradiated CV-1 monkey kidney cell monolayers was decreased by caffeine. X-ray enhanced reactivation of UV-irradiated virus in X-irradiated monolayers (X-ray reactivation) and UV- or X-ray-inactivated capacity of the cells to support unirradiated virus plaque formation were unaffected by caffeine. The results suggest that a caffeine-sensitive process is necessary for the expression of Weigle reactivation for herpes virus. Since cafeine did not significantly affect X-ray reactivation, different mechanisms may be responsible for the expression of Weigle reactivation and X-ray reactivation.


[Kinetic manifestations of the interaction of active centers in swine skeletal muscle D-glyceraldehyde-3-phosphate dehydrogenase].

The kinetic method and selective chemical modification have been used in studies of the kinetic manifestations of active site interactions in D-glyceraldehyde-3-phosphate dehydrogenase (GAP dehydrogenase). The reactions of glyceraldehyde and glyceraldehyde-3-phosphate oxidation were studied in the absence of substrate excess. In support of the data obtained previously it was shown that only a part of the tightly bound NAD molecules can be reduced after substrate addition. "Partial reducibility" is observed at various degrees of saturation of the enzyme with NAD involving a single NAD molecule per tetrametric enzyme. These facts can hardly be explained by assumption of functional non-equivalence of active sites, whether induced by coenzyme or preexisting in the apoenzyme. It was proven by selective alkylation of the catalytic SH groups that "partial reducibility" is due to the circumstance that equilibrium in the system under investigation is established at nearly equal NAD and NADH concentrations. A plot of initial reaction rates versus NAD concentration (at non-saturating substrate concentrations) gives S-shaped curves; this is explained by considerable enzyme activation upon saturation of the fourth site with coenzyme. After modification of three active sites with iodoacetate the S-shape of the curve disappeared. This fact leads to the conclusion that active site interactions are required for formation of the S-shaped curves. The activity of a single site functioning in the modified enzyme reached values equal to those of the active sites in the native enzyme in the fully activated state. A model is proposed which can explaine the variations in mode of enzyme activation in the native and modified states. It is suggested that the surroundings of all four SH groups must be altered in order to activate the enzyme; such changes can be induced either by alkylation of the SH groups or by NAD binding. Evidence is presented that important functional properties of GAP dehydrogenase cannot be elucidated at low enzyme concentrations and with excess of substrates: three active sites are saturated under such conditons and practically inactive, and the fourth site obeys Michaelis - Menten kinetics.


Infection of UV-irradiated xeroderma pigmentosum fibroblasts by herpes simplex virus: study of capacity and Weigle reactivation.

The capacity of monolayers of both normal human and xeroderma pigmentosum (XP) filbroblasts to support plaque formation by herpes simplex virus was decreased when the monolayers were ultraviolet (UV) irradiated and infected with virus. Fibroblasts of XP complementation groups A, B, and D were sensitive to UV, being 4-6 fold more sensitive than either fibroblasts of XP complementation group C or fibroblasts from a normal individual. When the monolayers were irradiated 4 days prior to infection, the capacity of normal fibroblasts to support herpes virus growth recovered, whereas the capacity of the XP strains decreased further compared to that measured when infection immediately followed irradiation. Concurrent experiments with UV-irradiated herpes virus showed that the survival of this virus did not increase when infection by irradiated virus immediately followed irradiation of the monolayers. However, if the monolayers were irradiated 4 days prior to infection, the survival of this virus increased by a factor of nearly 2. Such Weigle reactivation (WR) occurred at lower fluences to the XP fibroblasts than to normal fibroblasts, suggesting that WR results from residual cellular DNA damage left after excision repair.


Differential cytology in the diagnosis of astrocytary gliomas. A statistical study using the "t" Students-Fisher test.

The authors have used the "t" Student-Fisher test in order to verify the differences between two compared average values. The results show the anatomo-clinical individuality of four main types of astrocytary gliomas; multiform glioblastoma, malignant astrocytoma, protoplasmatic astrocytoma, and fibrillary astrocytoma. The occurrence of significant quantitative differences between these four astrocytary gliomas indicates that the histological diagnosis must be differentially made, because these astrocytary neoplasias represent individualized anatomo-clinical entities.


Inactivation and mutation of coliphage T4 by aliphatic nitrosamides and methanesulphonates: in vitro recovery of infectivity of T4 inactivated by isopropyl methanesulphonate.

The inactivation and mutation (to r phenotype) of extracellular coliphage T4 wild-type by the monofunctional alkylating agents N-methyl- and N-ethyl-N-nitrosourea and isopropyl methanesulphonate were investigated. The rate and extent of change in phage infectivity observed during the post-treatment period were found to correlate with what is known of the mechanisms by which these agents react in vitro. Loss of phage infectivity was found to occur during the period following treatment with these agents, but that resulting from treatment with isopropyl methanesulphonate was preceded, in the first 24 to 48 h, by a recovery of infectivity. This suggested that changes in phage infectivity occurring after treatment with monofunctional alkylating agents are resultant of various processes which diversely promote loss and recovery of infectivity. The mutagenicity of N-methyl-N-nitrosourea was similar to that of its ethyl homologue at a level of phage survival of 4 x 10-3, but less than that of isopropyl methanesulphonate. At a level of survival of 3 x 10-2 ethyl methanesulphonate was a mutagenic as its isopropyl homologue, but methyl methanesulphonate was only slightly if at all mutagenic. These results could not be correlated with the compounds' reaction mechanisms. The efficiency of isopropyl methanesulphonate (compared with its toxicity to phage) was found to decrease as the severity of the dose was increased.


Mutagenic and inactivating effects of methyl alkylaminosulfonates on Escherichia coli.

Methyl alkylamino methanesulfonates are mutagenic agents as shown by treating several strains of E. coli at pH 7. Methyl methylaminosulfonate (CH3-NH-SO3-CH3) was more efficient than methyl ethylaminosulfonate (C2H5-NH-SO3-CH3) which itself was more efficient than methyl isopropylaminosulfonate (C3H7-NH-SO3-CH3). Methyl methylaminosulfonate seemed to be at least as effective as methyl methanesulfonate (CH3-SO3-CH3). Methyl methylaminosulfonate produced a yield of up to 1% of auxotrophic mutants. All three new mutagens appeared to react according to the same mechanism by ester fission and methylation of nucleophilic groups as is known for methyl methanesulfonate. The reaction mechanism seems to be of the SN2 type.


Relationships between the changes of some oxidative and hydrolytic enzymes in the kidney of irradiated rat.

Changes of succinic dehydrogenase, adenosinetriphosphatase, alkaline and acid phosphatase in rat kidney, during 6 months after a whole-body 800 r X-ray exposure were studied. Temporary but long-lasting activation of the histoenzymatic reactions accompanied by partly reversible diffusion of enzymes related to the dystrophic and restoring postirradiation renal changes were observed.


Non-endemic Burkitts's lymphoma. A B-cell tumor related to germinal centers.

To investigate the nature of non-endemic Burkitt's lymphoma, we examined neoplastic cells from eight American patients for receptors for sheep erythrocytes (E), complement (EAC), and Fc fragment of lgG (igGEA), and for surface immunoglobulins (Slg) and hydrolytic enzymes. In addition, we reviewed 47 biopsies and 17 autopsies from American patients to ascertain patterns of involvement by tumor in lymph nodes, spleens and Peyer's patches. Neoplastic cells in all cases studies bore monoclonal surface immunoglobulins of the igM class. Receptors for EAC and igGEA were identified on a minority of the cells. Little or no hydrolytic enzyme activity was demonstrable. These results indicate that, like Burkitt's lymphoma in Africans, this histologically identical tumor in American patients consists of B lymphocytes. In 10 biopsies and two autopsies, germinal centers were selectively involved by tumor, suggesting that these neoplastic cells may be related to some B lymphocytes of normal germinal centers.


Distribution of antibody to hepatitis a antigen in urban adult populations.

To investigate the prevalence and distribution of antibody to hepatitis A antigen we tested 947 randomly selected people in the Greater New York City area; 45 per cent were antigen positive, as determined by the immune adherence method. Antibody was detected two to three times more frequently in lower social classes (72 to 80 per cent) than in middle and upper-middle classes (18 to 30 per cent). The rate of antibody detection was strongly correlated with age; the prevalence gradually increased throughout adulthood and reached its peak level in people 50 years of age and older. Those with serologic evidence of past exposure to hepatitis B virus were significantly more often antibody positive than those without such evidence (61 vs. 40 per cent; P less than 0.001). Very few of the positive subjects had had hepatitis. The prevalence of this antibody varies among different population groups, increases with age, decreases with rise in socioeconomic status, is independent on sex and race, and correlates with serologic evidence of hepatitis B virus infections.


A possible common mechanism of action of antidepressant treatments: reduction in the sensitivity of the noradrenergic cyclic AMP gererating system in the rat limbic forebrain.

The response of the noNEpinephrine (NE) sensitive cyclic AMP generating system in slices of the rat limbic forebrain after both the acute and chronic administration of the tricyclic antidepressants desipramine (DMI) and iprindole as well as electro-convulsive treatment (ECT) was investigated. Neither the basal level of cyclic AMP nor the hormonal response to NE were altered after administration of a single dose of short term treatment with DMI and iprindole. However, the administration of the antidepressants on a clinically more relevant time basis markedly reduced the sensitivity of the cyclic AMP generating system to NE. This change in sensitivity was not related to the levels of the drugs in brain. The response of cyclic AMP to NE was also reduced by ECT, but the onset of this action was shorter than that observed with the antidepressants. ECT also antagonized the enhanced response of cyclic AMP to NE following destruction of central adrenergic nerve terminals with 6-hydroxydopamine. It thus appears that the therapeutic action of tricyclic antidepressants could be related to postsynaptic adaptive changes in the sensitivity of the noradrenergic adenylate cyclase receptor system rather than to acute presynaptic events.


Possible role of cyclic AMP in the relaxation process of mammalian heart: effects of dibutyryl cyclic AMP and theophylline on potassium contractures in cat papillary muscles.

The effect of dibutyryl cyclic AMP (DB-c-AMP; 3 X 10(-4)-3 X 10(-3) M) on electrically induced twitch and high potassium (142.4 mM KCl)-induced contracture tension was studied in papillary muscles from normal and reserpinized cats ([Ca]0 1.8 mM; 25 degrees C; pH 7.4). In both groups of preparations, the increase in twitch tension evoked by DB-c-AMP was accompanied by an abbreviation of the time to peak force and of relaxation time. In the same preparations, the high potassium contracture was markedly depressed by DB-c-AMP in a concentration-dependent manner. Similar results were obtained with the N6-monobutyryl derivative of cyclic AMP. The relaxing effects of the cyclic nucleotides on KCl contractures did not appear to be due to possible non-cyclic breakdown products: adenosine, 5'-AMP and sodium butyrate did not attenuate contracture tension at concentrations up to 3 X 10(-3) M. The same applies to ATP and non-cyclic N6-2'-0-3'-0-tributyryl-adenosine-monophosphate. Theophylline (10(-2) M) was found to prolong the relaxation time of the twitch and to enhance the high KCl contracture. It is concluded that cyclic AMP may be capable of modulating the relaxation process  of mammalian heart and that not only the positive inotropic but also the relaxant effects of catecholamines on myocardium described before may be mediated by the cyclic AMP system. The relaxant effects of cyclic AMP derivatives on intact myocardial preparations are attributed to a stimulation by cyclic AMP of the calcium transport of the sarcoplasmic reticulum (SR) and are interpreted to be a corollary to the effects of cyclic AMP previously obtained on isolated SR preparations.


Antagonistic effects against single lethal doses of Amanita phalloides.

Agents with antagonistic effects against phalloidin or alpha-amanitin were tested in mice against lethal doses of an extract from the whole mushroom Amanita phalloides. The following categories of agents reduced lethality after the extract. First, agents protecting only against phalloidin such as rifampicin, phenylbutazone and antamanide. Second, silymarin and prednisolone which display both antiamatoxic and marked (silymarin) or moderate (prednisolone) anti-phallotoxic acitivty. Thioctic acid displayed some activity when tested against mid-lethal doses of the extract. Cytochrome c, a chemical with curative potencies against alpha-amanitin did not reduce the lethality of the exact. All of the effective agents acted only when applied prior to the poisoning. The pattern or protective activity would indicate that in mice death after single doses of Amanita phalloides may follow a qualitatively particular couse which is difficult to ascribe to phallo- or amatoxic effects alone.


Inhibition by polyhalogenated hydrocarbons (PHHC) of ATPases in plasma membranes of parenchymal liver cells.

Suspensions of freshly prepared plasma membranes from rat livers were equilibrated with defined mixtures of air with polyhalogenated hydro-carbons (PHHC) at 37 degrees C. During the exposition enzymatic degradation of ATP was inhibited by carbon tetrachloride, chloroform and halothane in a dose-dependent manner. On a molar basis carbon tetrachloride was 5 times more effective than chloroform and halothane. The inhibition was partially reversible. Diethyldithiocarbamate, an agent protecting animals against carbon tetrachloride, was ineffective in this assay.


Effects of adenosine on adrenergic neurotransmission; prejunctional inhibition and postjunctional enhancement.

The action of adenosine on adrenergic neuroeffector transmission was studied in the rabbit kidney in vitro and in situ, in the canine subcutaneous adipose tissue in situ and in the guinea pig vas deferens in vitro. In the kidney, adenosine (0.1-10 muM) caused a concentration-dependent increase in vascular resistance and in vasoconstrictor responses to nerve stimulation and administered noradrenaline. In the adipose tissue, adenosine also increased the vaso-constrictor responses but it decreased vascular resistance. In all three tissues studied adenosine significantly and reversibly depressed noradrenaline release evoked by nerve stimulation in a concentration-dependent manner. This effect of adenosine was not altered by phenoxybenzamine which blocked all vasoconstrictor responses and diminished the rise in vascular resistance by adenosine in the kidney. It is concluded that adenosine affects adrenergic neuroeffector transmission by two discrete mechanisms, prejunctional inhibition and postjunctional enhancement.


Immunodiffusion analysis of the antigenic structure of the transplantable MC 29 tumor.

By the use of specific anti-chicken liver sera it could be proved with immunodiffusion techniques that the transplantable MC 29 tumor line contains liver-specific antigens. Production of chicken serum-proteins by the tumor could also be demonstrated. Thus, the transplantable MC 29 tumor line can be regarded as a hepatoma.


Differentiation of intestinal smooth muscle relaxation caused by drugs that inhibit phosphodiesterase.

In this study a number of chemically unrelated smooth muscle relaxants were tested: a) for potency of phosphodiesterase (PDE)-inhibition, using guinea pig colon-PDE and rat erythrocyte-PDE, b) for potency and duration of relaxation of the isolated guinea-pig colon, c) for their interaction with NH4Cl and orciprenaline as well as with "high calcium". Compared with the spasmolytic effect inhibition of guinea pig colon-PDE or rat erythrocyte-PDE was strong for papaverine and some other relaxants but was low or virtually absent with verapamil, hexobendine and bencyclane. The spasmolytic effect of some PDE-inhibitors was found diminished by the PDE-activator NH4Cl. Most of these drugs enhanced the relaxant effect of the "cyclase activator" orciprenaline; the latter are designated A-type drugs. Verapamil, bencyclane, hexobendine and M 13 did not show this type of interaction with NH4Cl and orciprenaline; they are designated B-type drugs. With A-type drugs--but not with B-type drugs--a highly significant correlation was observed between potency of relaxation and inhibition of colon-PDE (r = 0.85) as well as rat erythrocyte-PDE (r = 0.77). The spasmolytic action of aminophylline and papaverine (A-type drugs) was not inhibited by elevation of extracellular calcium to 9 mM, whereas relaxation induced by verapamil and hexobendine (B-type drugs) at 1.8 mM calcium was found abolished by 9 mM calcium. It is concluded that A-type drugs relax guinea-pig colon by inhibition of PDE and accumulation of cAMP, and that B-type drugs in all probability act by (a) cAMP-independent mechanism(s).


Transplantability and biological behavior of a rapidly growing secondary Morris hepatoma.

Transplantable hepatomas have a special importance in cancer research because most are well-differentiated and grow at widely differing rates. A simple and reproducible technique was developed to transplant the rapidly growing Morris hepatoma 9618A2 from a subcutaneous to an intrahepatic site. The tumor has been maintained for over 80 successive transplant generations by inoculating Buffalo rats subcutaneously. The hepatoma, transplanted many times from the subcutaneous site to the liver, was examined histologically on various occasions. By varying inoculum size, growth of the hepatocarcinoma could be controlled. Throughout transplantation, the donor animals were bled without causing the death of the animal. Histological studies revealed that the tumor, when transplanted intrahepatically, remains a highly differentiated neoplasm with no observable morphological changes.


Adrenocortical responses of rats to acute hypoxic and hypercapnic stresses after treatment with aminergic agents.

Monoaminergic influences on the regulation of the hypothalamo-hypophyseal-adrenocortical (HHA) system during acute stresses (hypoxia and hypercapnia) were investigated in male rats. Plasma corticosterone levels were used to assess HHA activity, and the alterations in monoaminergic activity were induced by pretreating the animals with various pharmacologic agents (reserpine, alpha MT, FLA-63, pCPA, L-Dopa, pargyline, Lilly 110140, phentolamine and propranolol). Dexamethasone-treated rats were utilized to assess the site at which these monoaminergic substances acted. The latter experiments showed that these agents did not have a marked effect directly on the adrenal cortex and thus the site(s) of action was at the level of the anterior pituitary and/or above. Altering the serotoninergic system did not appreciably influence the HHA response to hypoxia and hypercapnia, whereas increasing the activity of the adrenergic system partially prevented the rise usually observed in plasma corticosterone levels during these stresses. These data suggest that different aminergic pathways may be utilized for different stresses.


Injection of melatonin into cisterna magna increases concentration of 3', 5' cyclic guanosine monophosphate in cerebrospinal fluid.

Rabbits were anesthetized with urethane and were given intracisternal injections of the following substances: adrenocorticotropin, beta-melanocyte stimulating hormone, choroid plexus peptide IIF, epinephrine, serotonin, histamine, oxytocin, lysine and a arginine vasopressins, acetylcholine and melatonin. The effects on the concentration of 3', 5' cyclic guanosine monophosphate (cGMP) in cerebrospinal fluid were then measured. Only melatonin and acetylcholine caused a significant (p less than 0.05) effect on cGMP concentration. Both agents increased the nucleotide's concentration within 30 min. Melatonin was about 1,000 times more potent than acetylcholine; the mininal effective doses were 1 mug and 1,000 mug, respectively.


[Serum lipids in Down's syndrome].

In 28 patients with Down syndrome and in 28 controls matched for age and sex serum lipoproteins and their main lipid constituents were determined. Both groups comprised subjects living in the same institutions for mentally retarded No significant differences were found between the two groups in total serum lipids, triglycerides, total cholesterol and phospholipid levels and in the proportion of esterified cholesterol and beta-lipoproteins. No characteristic pattern of alpha and beta lipoproteins could be observed in patients with Down syndrome by means of electrophoresis in polyacrylamide gel. Alanine aminotransferase activity was significantly higher in the group of patients.


Sleep EEG and motor activity as indicators in affective states.

This report reviews a number of studies which support our current classification schema for affective disorders. This classification differentiates between an anxious-hyperactive type and an anergic-hypoactive type which are then further subdivided into primary and secondary affective disorders. While the motor and activity measurements are for the most part limited to patients suffering from primary affective disease, current studies under way indicate that secondary affective disorders may also have characteristic biologic changes. EEG sleep and motor activity parameters provide useful pointers for differential diagnosis and treatment while having the added advantage of diminishing the clinician's almost exclusive dependency on amnestic data and the psychological observations made on the patient's behavior.


The effect of ACTH treatment on lymphocyte subpopulations in patients with myasthenia gravis.

Blood lymphocytes were collected before, at various times during treatment and after treatment with ACTH in patients with myasthenia gravis. The relative proportions of thymus-derived (T cells) and bursa-derived (B cells) lymphocytes were determined. Eight of nine patients had a relative increase in B cells, whereas T cells showed a relative decrease. Myasthenic symptoms in these eight patients also were relieved. One patient who was not thymectomized showed virtually no relief in symptoms nor any changes in the relative T and B cell proportions or numbers, indicating a close association between the disease and the thymus-derived immune system.


"Giant axonal neuropathy" caused by industrial chemicals: neurofilamentous axonal masses in man.

Symmetrical polyneuropathy developed in two patients after they had been in contact with acrylamide and methyl n-butyl ketone, respectively. In sural nerve biopsy material from both patients, electron microscopy showed frequent focal axonal swellings containing masses of neurofilaments. Some axons undergoing axonal degeneration also were seen. These morphologic features are identical to those produced in experimental animals after exposure to these chemicals and are similar to those found in n-hexane neuropathy and in the three reported cases of giant axonal neuropathy. Sural nerve biopsy is an important diagnostic test in identifying cases of peripheral neuropathy caused by these chemicals.


Hereditary sensory neuropathy and tonic pupils.

The clinical features of two siblings with both hereditary sensory neuropathy and bilateral tonic pupils are described. Other neurologic disorders associated with tonic pupils are reviewed. Our patients appear to represent a unique instance of the association of disorders that primarily affect the ciliary and dorsal root ganglia.


The childhood type of dermatomyositis.

The childhood type fo dermatomyositis, which occurs in children and young adults, shows a specific constellation of pathologic changes in muscle. Capillary necrosis leads to capillary loss, generally starting on the periphery of muscle fascicles. Electron microscopy discloses undulating tubules in endothelial cells, lymphocytes, pericytes, and pseudosatellite cells. The muscle fiber damage is coextensive with capillary damage and probably results from progressive ischemia. The muscle cells, before atrophying, show mitochondrial elongation, Z disk streaming, focal myofibrillary loss, and occassionally selective thick filament loss. Muscle cell necrosis is rare and limited to infarctlike lesions. Inflammatory infiltrates, if present, occur only in connective tissue septa. The cause of the capillary damage has not been determined.


Peripheral motor and sensory neuropathy of early childhood, simulating Werdnig-Hoffmann disease.

A five-year-old girl developed neurogenic muscular atrophy during infancy. On the basis of clinical findings and a muscle biopsy study a diagnosis of infantile spinal muscular atrophy, benign type II, was entertained. As the disease progressed, involvement of sensory pathways was detected by electrophysiological studies. Examination of a sural nerve specimen suggested a chronic polyneuropathy rather than spinal muscular atrophy. These findings were confirmed at autopsy by demonstrating severe loss of myelinated axons in ventral and dorsal spinal roots, peripheral nerves and dorsal columns of the spinal cord. In addition, islands of astroglial fibers were found in ventral and dorsal spinal roots, regarded as a secondary reaction to the breakdown of myelinated axons. This unusual scarring process seems to result from nerve fiber loss during the perinatal period, since radicular glial scar tissue is not known to occur in the spinal muscular atrophies of later onset. Examination of the sensory nervous system in patients with Werdnig-Hoffmann disease appears mandatory to clarify the precise disease entity leading to infantile neurogenic muscular atrophy.


The diagnosis of infantile generalized ceroidlipofuscinosis (type Hagberg-Santavuori) using skin biopsy.

Skin biopsies were performed in two cases of infantile generalized ceroidlipofuscinosis. In the first case the biopsy was done at the age of 8 years and the diagnosis was confirmed shortly thereafter by a postmortem examination. In the second case, the biopsy was performed much earlier (27 months) at a time when the diagnosis was not suspected; it was subsequently confirmed by similar findings in a neuro-muscular biopsy. In both cases, numerous cytoplasmic inclusions with granular osmiophilic deposits were present in epidermal cells, eccrine sweat glands, smooth muscle cells, nurve fascicles, fibroblasts and vascular elements while none were found in age-matched controls. As already stressed by Anzil et al. (1975), the morphology of the inclusions can be heterogeneous and linear profiles were often observed, mainly in the cytosomes present in the vascular cells. We feel confident that a diagnosis of infantile generalized ceroid-lipofuscinosis can be made by skin biopsy obviating therefore the need for other surgical procedures.