Anticellular effects of 9-(2-hydroxyethoxymethyl) guanine against herpes simplex virus-transformed cells.

The guanine derivative 9-(2-hydroxyethoxymethyl) guanine (acycloguanosine), a potent inhibitor of herpes simplex virus (HSV) multiplication, was found to have marked anticellular activity against HSV-transformed thymidine kinase-positive cells. HSV type I-transformed cells were more sensitive to the drug than HSV type 2-transformed cells.


Inactivation of human cytomegalovirus by the chemical carcinogen 4-nitroquinoline 1-oxide.

The infectivity of cytomegalovirus (CMV), strain Davis, was inactivated by 4-nitroquinoline 1-oxide (NQO). A series of survival curves indicates that the rate of inactivation was directly dependent on the concentration of NQO over a range of 5 to 200 microgram/ml. At concentrations of I microgram/ml or less, inactivation of virus stock was not observed and at concentrations in excess of 200 microgram/ml, the cellular toxicity of residual NQO prevented quantification of the relatively low surviving infectivity. At a concentration of 200 microgram/ml NQO or less, the loss of virus infectivity could be clearly shown to result from the interaction of NQO with virus and not with cells, since the addition of similar doses of NQO to assay cells simultaneously with virus did not adversely affect the sensitivity of the assay cells to measure virus infectivity. Similarly, the dimethylsulphoxide carrier at concentrations of 5% or less was shown to have a negligible effect on both virus infectivity and on the sensitivity of human skin muscle cells to assay virus infectivity. NQO inactivation of virus infectivity appeared to depend very little on white light, since the kinetics of inactivation in the presence and in the absence of white light were similar.


Absence of significant antiviral effects of beta-gamma-methylene GTP on encephalomyocarditis virus infection of L cells and mice.

The protein synthesis inhibitor beta-gamma-methylene guanosine triphosphate (Gpp-CH2p) is shown here to be ineffective as a 'leaky membrane' antiviral agent against encephalomyocarditis virus infection of L cells and mice. Studies with GppCH2p in encephalomyocarditis virus-infected L cells indicate that the cells only become permeable to the inhibitor late in infection because the compound significantly inhibits protein synthesis only when added at 4 h p.i. At this time 50 to 70% of the new infectious virus particles have already been synthesized, and this is reflected in maximum inhibition of virus yields of only about 40%. Moreover, comparison of inhibition of protein synthesis by GppCH2p in vitro and in cell cultures indicates that the intracellular concentration attained is only 0.25% of that in the medium. The lack of antiviral activity of GppCH2p in encephalomyocarditis virus-infected mice is probably due to leakiness of infected cells occurring too late for sufficient inhibition of virus synthesis to be obtained.


Protease activity on the surface of HSV-infected cells.

Monolayers of primary rabbit kidney cells infected with HSV type I bound lymphoblastoid (Raji) cells, to which the third component (C3) of the complement system had been attached (Raji-C3). This induction of cell contact did not occur on non-infected monolayers and was dependent on C3. The interaction could be suppressed by the presence of protease inhibitors (1 mM-TLCK or-PMSF). The results are interpreted as indicating de novo expression of protease activity on the surface of HSV-infected cells. This protease is characterized by its potential to activate C3 and its inhibition by TLCK.


Studies on antiviral glycosides, 4. Inhibition of the multiplication of paramyxoviruses by phenyl-6-chloro-6-deoxy-beta-D-glucopyranoside.

The antiviral activity of phenyl-6-chloro-6-deoxy-beta-D-glucopyranoside (PCG) was studied. PCG specifically inhibited the growth of paramyxoviruses including Sendai, measles and Newcastle disease viruses in LLCMK2 cells at a concentration of 0.5 to 1.0 mM, but did not restrict the multiplication of other RNA viruses (influenza, vesicular stomatitis and polio viruses) at these concentrations. PCG might act in the late stage during virus replication of Sendai virus as it did not inhibit virus RNA and protein synthesis in the infected cells. Comparative studies on the biological properties of virus particles grown in the presence and absence of PCG demonstrated that treatment with it caused the formation of non-haemagglutinating particles.


Detection of virus antigens in Swiss albino mice infected by milk-borne mouse mammary tumour virus: the effect of age, sex and reproductive status. I. Localization by immunofluorescence of four antigens in mammary tissues and other organs.

The indirect immunofluorescence technique was used to investigate the expression of virus antigens in a Swiss albino mouse strain infected by mouse mammary tumour virus (MMTV). The antisera used were monospecific for: the glycopolypeptides gp160 and gp47, and the internal polypeptides p28 and p8. In mice infected with milk-borne MMTV, all four antigens were detected in the mammary gland during the first pregnancy and then throughout life, and in mammary tumours. Antigen gp160, located at the secretory border of all glandular cells, is shared by the plasma membrane of the mammary cell, whether virus-producing or not, and by the virus envelope, as shown by the use of absorbed antisera. The other three antigens are virus-specific. Anti-p47 serum stained the secretory border of cells whereas the fluorescence related to p28 and p8 consisted of a few large intracytoplasmic spots. The specific antibodies for p28 and p8 were only absorbed by disrupted virions, confirming that these polypeptides are internal antigens. With serial sections of glands from pregnant and old female mice, the alveoli stained with anti-p28 or anti-p8 serum were found to react also with anti-gp47 serum. However, during lactation, the presence of p28 and p8 could not be detected in the alveoli stained with anti-gp47 serum. Thus, infected alveoli express virus antigens differently. In the mammary glands of all mice studied, some alveoli did not display any staining when the latter three antisera were used. The specific antigenic pattern, maintained in differentiated mammary tumours, became a diffuse and unlocalized staining after transformation into the undifferentiated state. With the exception of the kidney, where gp47 was found in the glomeruli, all other organs of the mouse did not stain with any of the antisera. The presence of gp47 in the glomeruli, probably related to immune-complex deposits, increases with the age of the mouse and is most noticeable in tumour-bearing females. In mice free of milk-borne MMTV, examined during the first week of lactation, antigen gp47 could not be detected. Internal antigens p28 and p8 were detected in nearly all mammary cells as numerous small cytoplasmic dots. This suggests a limited expression of an endogenous virus genome; this expression would be modified when milk-borne virus is produced.


Detection of virus antigens in Swiss albino mice infected by milk-borne mouse mammary tumour virus: the effect of age, sex and reproductive status. II. Radioimmunoassay of two virus components, gp47 and p28 in serum and organ extracts.

Extracts of various organs, mammary tumours and sera from milk-borne MMTV infected Swiss albino mice of different age, sex and physiological conditions were tested by radioimmunoassay for the presence of gp47, the main envelope polypeptide, and p28, the main core protein of the virus. Except in brain, ovaries and testes, both antigens were found in all organs of old animals and of females after the onset of their first pregnancy. Antigens were not present in organs of weanlings or in whole foetuses. Higher values were found in mammary glands, mammary tumours, epididymis and seminal vesicles. These organs also harboured a greater amount of gp47 than p28. The serum generally contained gp47 but rarely p28. This indicates that gp47 is not virion-bound in blood. Pregnancy, lactation and especially the presence of mammary tumours increased the concentration of gp47 in serum. The results do not allow localization of target organs of MMTV infection in the interval between ingestion of the virus by the suckling mouse and the first pregnancy. Moreover, results obtained with one group of mice devoid of exogenous virus show that, as endogenous MMTV genome expresses p28, it might account for part of the p28 detected exogenous MMTV-infected mice.


Cell cycle-dependent chronic infection of human cytomegalovirus in human osteogenic sarcoma cells.

Human osteogenic sarcoma cells transformed by murine sarcoma virus (R970) showed restricted growth of human cytomegalovirus (HCMV). Virus was attached to the same extent as in human fibroblasts. HCMV growth was blocked at early stages after virus penetration. Splitting of infected R970 cultures resulted in infection of all cells. In experiments using synchronized R970 cells it was found that factor(s) associated with the S-phase of the cell cycle might be necessary for establishment of the infection.


Selective replication of transformation-defective avian sarcoma virus mutants in duck embryo fibroblasts.

When an avian sarcoma virus (ASV), subgroup C Bratislava 77 (B77-C) was inoculated into duck embryo fibroblast cultures (DEF) at a m.o.i. of 0.02, its replication was retarded by about 3 days compared with that in chick embryo fibroblast cultures (CEF). A transformation-defective (td) mutant was isolated during this period of retardation. Unlike the sarcoma virus, this td mutant replicated in both DEF and CEF with no retardation, even at a low m.o.i. The subgroup C Prague strain of Rous sarcoma virus (PR-C), which can infect DEF, also replicated in DEF slower than its td mutant, tdPR-C, at a m.o.i. of 0.02.


Synaptic connections and functional organization in Aplysia buccal ganglia.

The buccal ganglion of Aplysia contains three morpho-functional groups (A, B, and C) of large cells and two groups (s1 and s2) of small cells. The A cells evoke monoxynaptic IPSPs in the B cells. We found that s1 cells can evoke large EPSPs in the A cells, IEPSPs in the B cells, and EIIPSPs in the C cells; several s1 cells are able to evoke all three types of responses. Many s2 cells can evoke these same responses, but only in the A and B cells. Furthermore, the s cells can evoke depolarizing PSPs in other s cells; this relation is often reciprocal. All these responses may also be contralateral. Their monosynaptic nature is shown by the consistent 1:1 relationship with the presynaptic spike, and also by the effects of intracellular tetraethylammonium and of high Mg2+ concentration in the bathing medium. d-tubocurarine reversibly suppresses the I phase of the IEPSP evoked by the s cells in the B cells. All the responses evoked by the s cells undergo depression with repetition. The network formed by all these relations is outlined, and a double relationship proposed between s cells and B cells. By electrophysiological tracing of axonal pathways it is shown that the A cells send axons into the 3rd buccal nerve, the B cells into the 2nd and/or 3rd buccal nerve and in two cases into the radular nerve, and the C cells into the gastro-oesophageal nerve. Spontaneous synaptic activity of the buccal neurons appears to be formed mostly by the described PSPs. Spontaneous firing inside the isolated ganglion corresponds well to the alternate pattern of muscular contractions of the buccal mass.


Effects of amino acids on calcium uptake by glial and neuroblastoma cells.

The uptake of [45Ca] has been studied in clonal glial and neuronal cells. It was somewhat more efficient in the neuroblastoma clone M1 compared to glial clones. In all cases [45Ca] uptake was shown to depend on the phosphate concentration in the incubation medium. It was decreased by the ionophore A 23187  at 200 microM concentration in both neuronal and glial clones. The influence of amino acids some of which are putative neurotransmitters was investigated; the interactions between [45Ca] uptake and these amino acids were related to their concentration and the type of cells used (neuronal or glial). L-aspartate and taurine for example had two opposite effects on [45Ca] uptake by the glial clone NN at two different concentrations; they could therefore play a role in the control of calcium level in the synaptic cleft.


Mg++-sensitivity of neuromuscular transmission in two crustaceans: correlation with blood Mg++ levels.

Neuromuscular transmission was measured in muscles of spider crabs (Hyas areneus) and lobsters (Homarus americanus). Solutions containing 40 and 10 mM/1 Mg++, which were approximately the same as those measured in the blood of Hyas and Homarus, respectively, were used to soak the preparations prior to testing. In Homarus, neuromuscular transmission was severely depressed by 40 mM Mg++. In spider crabs, neuromuscular transmission was not severely depressed. Although the amount of transmitter released by nerve impulses was reduced, total membrane depolarization during trains of impulses was not reduced because a compensating increase in muscle fiber membrane resistance occurred in Hyas preparations exposed to M Mg++. Hyas, but not Homarus, is physiologically adapted to function at relatively high blood Mg++ concentrations.


Neurotoxicity of halogenated hydroxyquinolines: clinical analysis of cases reported outside Japan.

An analysis is presented of 220 cases of possible neurotoxic reactions to halogenated hydroxyquinolines reported from outside Japan. In 80 cases insufficient information was available for adequate comment and in 29 a relationship to the administration of clioquinol could be excluded. Of the remainder, a relationship to clioquinol was considered probable in 42 and possible in 69 cases. In six of the probable cases the neurological disturbance consisted of an acute reversible encephalopathy usually related to the ingestion of a high dose of clioquinol over a short period. The most common manifestation, observed in 15 further cases, was isolated optic atrophy. This was most frequently found in children, many of whom had received clioquinol as treatment for acrodermatitis enteropathica. In the remaining cases, a combination of myelopathy, visual disturbance, and peripheral neuropathy was the most common manifestation. Isolated myelopathy or peripheral neuropathy, or these manifestations occurring together, were infrequent. The onset of all manifestations (except toxic encephalopathy) was usually subacute, with subsequent partial recovery. Older subjects tended to display more side effects. The full syndrome of subacute myelo-optic neuropathy was more frequent in women, but they tended to have taken greater quantities of the drug.


Soleus motoneurone excitability in man: an indirect approach for obtaining quantitative data.

Using combinations of H and T reflexes, spatial summation has been studied in the human soleus motoneurone pool. With certain assumptions, mathematical treatment of the results yielded estimates of the thresholds of motoneurones to monosynaptic activation by Ia nerve fibres. It was found that, on average, about 62 EPSPs were required to discharge a motoneurone and that a single Ia fibre supplied about 139 neurones.


Neuromuscular transmission in thyrotoxicosis.

The occurrence of thyrotoxicosis in myasthenia gravis is estimated as 5.3% (Osserman et al. 1967) but the incidence of myasthenia in the course of thyrotoxicosis is only 0.2% (Simpson 1968). The association between these two diseases is not a chance one and there are two possible basic mechanisms viz. the action of thyroid hormone on the neuromuscular junction or the existence of a cause common to both. The fatigability seen clinically in thyrotoxic myopathy could be partly attributed to myasthenia. With these possibilities in mind, a search for electromyographic stigmata of latent myasthenia was done on 48 consecutive cases of hyperthyroidism who also had chronic myopathy by performing repeated nerve stimulation. Eight cases had abnormal neuromuscular transmission as in myasthenia gravis and one of these cases later developed clinical myasthenia involving the ocular muscles. Three other cases showed abnormal facilitation as in the Eaton-Lambert syndrome.


A spin label study of myelin fluidity with normal and pathological peripheral nerves.

The myelin fluidity in peripheral nerves from normal subjects and patients with neuropathy has been studied. Nerve biopsies were made in subjects without neuropathy and in patients with Guillain-Barré, alcoholic or diabetic polyneuropathies and heredodegenerative diseases. The fluidity was studied using a stearic acid spin label with a nitroxide radical at C12 (12-doxylstearic acid) which was introduced by exchange between spin label-charged BSA and nerve myelin. Spectra was obtained with a VARIAN E 109 ESR spectrometer, at temperatures from 5 to 45 degrees C. The ultrastructure of biopsied nerves was also studied. The electron spin resonance (ESR) spectra were all typical of a partially immobilized nitroxide with a distance between outer hyperfine lines (2T//) of about 50 G, at room temperature. The curve 2T// against temperature shows two critical temperatures: 10 and 30 degrees C for all nerves. No correlation could be established between ultrastructure and fluidity of nerves. However, the myelin of nerves from diabetic patients showed a significant increase in viscosity. This physicochemical modification may represent an initial alteration of the myelin sheath, and demands further investigation. No modification of the myelin fluidity was observed in any of the other nerves studied.


Calcium deposits in the perineurium and their relation to lipid accumulation.

Calcification of the perineurium in sural nerves occurs in human adults. Deposits of calcium salts can be found focally in the middle and outer layers of the perineurium. Evidence indicates that this calcification is related to nerve fibre breakdown and to the local presence of lipids in the perineurium. The origin of the lipids may be senescent cells or degradation myelin.


Cyclic nucleotide phosphodiesterase activity in muscle of patients with carcinoma.

Cyclic nucleotide phosphodiesterase activity has been measured in muscle biopsies taken from healthy controls and from cancer patients. In both groups the muscles were clinically and morphologically normal. The phosphodiesterase activity was significantly increased in muscles from cancer patients using both cyclic AMP and cyclic GMP as substrate. These findings are in line with previous reports indicating that malignancy may interfere with metabolism of the host muscular tissues, and suggest the possibility that the observed biochemical changes might be an aspect of an early muscle neurogenic involvement.


Femoral neuropathy due to compression by retroperitoneal haemorrhage: a modern evaluation.

We report a case of femoral neuropathy due to retroperitoneal haemorrhage occurring during the administration of heaprin. The site of the hematoma is illustrated in the CT scan. It is emphasised that the modern assessment of peripheral neuropathies in the lower limbs associated with retroperitoneal haemorrhage by means of CT scan will promote our understanding of the natural history of this condition.


Changes in growth and morphology of human gliomas and fibroblasts cultured in D-valine medium.

Human fibroblasts and cells cultured from a glioblastoma multiforme and and oligodendroglioma were grown in standard tissue culture medium and in a medium in which D-valine was substituted for L-valine (DVM). Growth of all three types of cells was inhibited in DVM. Morphological changes of cells cultured in DVM also occurred: many fibroblasts became stellate-shaped and the oligodendroglioma cells had fewer processes. These findings indicate that DVM can not be used to decrease fibroblastic contamination in cultures of human gliomas without altering the morphological and growth characteristics of the neoplastic cells.


Topography of Pick inclusion bodies in hippocampi of demented patients. A quantitative study.

Topographic analysis was performed on the distribution of argyrophilic inclusions in hippocampal neurones of patients with Pick's dementia. A semiautomated scanning stage microscope linked potentiometrically to an XY pen recorder permitted the plotting of cytoarchitectonic "scattergrams" from the sequentially screened hippocampal formations. The density of Pick body-bearing nerve cells per cubic mm. of (pyramidal) cortex was quantified by measuring the area of each of six "zones" with a digitizer and programmable calculator. The ranking orders of relative severity showed that neurones in Rose's H1 field, the adjacent subiculum, and the entorhinal cortex are severely involved; that H2 and the presubiculum are less afflicted; and that the endplate is the least affected of all. The similarity of these predilections to those already demonstrated for Hirano bodies, granulovacuoles and neurofibrillary tangles in Alzheimer's disease suggests that a specific neurotransmitter defect may also underlie the dementia of Pick's disease.


The CO2 laser beam in the surgical treatment of cerebral tumours.

Several cases of supratentorial gliomas and meningiomas were operated on since May 1977 with the help of CO2 laser infrared beam, generated by the Sharplan 791 He unit, with an adjustable output up to 50 Watts. Defocused laser beam has been found to be useful in direct attack on the tumour, through a vaporization of the neoplastic tissue. So the tumour could be progressively excavate with a loss of blood comparatively lower than with classical method. Haemostasis was quite good on diffuse capillary bleeding, but not on arterial or venous haemorragies, where the use of electrocoagulation was necessary. Extirpation especially, of meningiomas, was easier and less traumatic with the laser beam, and the destruction of infiltrating glial tumours was facilitated too.


The demonstration of alpha lactalbumin in human breast carcinomas.

Alpha lactalbumin, a good marker of functional differentiation, has been demonstrated in 51 of a 100 human breast carcinomas examined using the immunoperoxidase technique. There is no relationship between production of alpha lactalbumin and histological differentiation, and the absence or presence of lymph-node metastases.


The nature of virus-like particles in the paraxial muscles of idiopathic scoliosis.

Virus-like particles (VLP) have been identified by electronmicroscopy in the skeletal muscles (paraxials) of six cases of idiopathic scoliosis. These particles closely resembled VLP reported in the skeletal muscles in other conditions, e.g. Reye's syndrome, polymyositis, malignant hyperthermia, and chronic myopathy. We have shown by specific staining that these structures are composed of glycogen in a crystalline form. Using Coxsackie B infected tissue culture cells as a control we have shown that these viruses, which are of similar shape and size to the VLP, were unstained using this specific staining method.


Activation by sanguinarine of active sodium efflux from frog skeletal muscle in the presence of ouabain.

1. Applied to intact Na-rich muscle cells, sanguinarine causes an increased 22Na efflux in the presence or absence of extracellular K+ or of ouabain. 2. The increased 22Na efflux does not represent Na:Na exchange as indicated by the fact that it is not associated with an increase in one-way isotopic Na influx nor is it abolished by the absence of external Na+. 3. In both K-free Ringer and K-free Ringer containing ouabain, sanguinarine not only increases one-way efflux of 22Na, it also induces net efflux of Na+ in the face of both an electrical and a concentration gradient. Moreover, the induction of net Na+ efflux occurs in the face of an approximately fourfold increase in PNa. These surprising results lead to the conclusion that, contrary to all experiments, sanguinarine induces active Na+ efflux even in K-free Ringer containing 10(-3) M-ouabain. 4. Sanguinarine depolarizes the Na-loaded muscle to approximately the same value, -54 mV, regardless of the presence or absence of extracellular K+. This depolarization is most likely secondary to the increase in PNa. 5. Sanguinarine causes a net loss of K+, presumably secondary to the depolarization. 6. The stimulation of net Na+ efflux is not correlated with the depolarization. The stimulation in K-free conditions (with or without ouabain), which is associated with the largest depolarization, produces an increment in net Na+ efflux which is not significantly different from the increment in net Na+ efflux in 10 mM-K+ Ringer where the depolarization is smallest. 7. Although sanguinarine increases active Na+ efflux in intact cells, it inhibits the isolated (Na+ + K+)-ATPase, presumably due to interaction with a site on the inner face of the membrane fragment. 8. The surprising stimulation of active Na+ efflux in the presence of 10(-3) M-ouabain must be due to interaction of sanguinarine with a site on the outer face of the membrane, perhaps the K+ activation site. It seems probable that the component of active Na+ efflux induced by sanguinarine is mediated by the Na pump. Sanguinarine may produce a K+-like effect upon the Na pump with consequent unbinding of ouabain.


Cyclic GMP and the permeability of the disks of the frog photoreceptors.

1. The diffusion of sodium, potassium and rubidium (not chloride) ions across the disk membrane is increased by cyclic guanosine monophosphate (cyclic GMP). 2. The increase is greater for sodium than for rubidium in the 0.01-0.1 mM concentration range. 3. Cyclic adenosine monophosphate (cyclic AMP) is less efficient than cyclic GMP; GMP and guanosine triphosphate are without effect. 4. The effect is present with either 1.8 mM calcium ions or 4 mM-EGTA in the perfusion fluid. 5. The presence of the cyclic GMP phosphodiesterase on the disk membranes is not needed for this effect. 6. The effect is present in both unbleached and fully bleached membranes.


Nephron electrolyte transport and sodium-potassium adenosine triphosphatase activity: influence of nicotine in rat and rabbit.

1. The influence upon mammalian renal epithelial transport of L-nicotine was studied in different types of experiments. 2. The kidney in situ (rat), when infused with nicotine (1 mg hr-1 kg-1 I.V.), lowered its absolute and fractional K+-absorption significantly and reversibly, but Na+-absorption did not change. Effects on glomerular function and an irreversible effect upon epithelial Na+ and K+ absorption were prevailing at higher infused amounts (10 or 50 mg hr-1 kg-1). 3. Single dissected nephron segments (collecting tubule, rabbit) were perfused in vitro, and the Na+ and K+-transtubular net flux was measured while L-nicotine (50 ng/ml.) had been added to the contraluminal side of the epithelium. Both, Na+-absorption and K+-secretion were decreased reversibly. 4. The activity of the Na+-K+-activated ATPase was significantly decreased in the cortical collecting tubule and in the proximal convoluted tubule of the rabbit after incubation of single in vitro dissected nephron segments with L-nicotine (50 or 100 ng/ml.). In contrast, nicotine added to a homogenate of renal cortical tissue had no effect on the Na+-transport enzyme or on key enzymes of glycolysis and gluconeogenesis. 5. These observations on the kidney in situ and on defined, perfused and non-perfused nephrons in vitro suggest that L-nicotine has dose-dependent, direct epithelial and mediated systemic effects on mammalian renal ion transport.


Retinal varicosity in Klippel-Trenaunay syndrome.

A retinal varicosity in the Klippel-Trenaunay syndrome is observed and diagnosed in a 24-year-old female. Fluorescein angiography was used to illustrate and specify the nature of the retinal vascular anomaly.


Structure--activity relationships of pyrrole amidine antiviral antibiotics. 1. Modifications of the alkylamidine side chain.

Representatives of three types of side-chain analogues of distamycin A (1) were synthesized. These were tested for cytotoxicity, inhibition of herpes simplex virus (HSV) replication in cultured cells, effects on the synthesis of HSV DNA in isolated nuclei in vitro, as well as on DNA synthesis by purified HSV DNA polymerase. Distamycin A was the most active compound in all three antiviral tests, as well as the most toxic. However, several compounds, in particular the aromatic analogues 15 and 16, showed no toxicity under the experimental conditions used but were still very active in the three antiviral tests.


A model of Martian surface chemistry.

Alkaline earth and alkali metal superoxides and peroxides, gamma-Fe2O3 and carbon suboxide polymer are proposed to be constituents of the Martian surface material. These reactive substances explain the water modified reactions and thermal behaviors of the Martian samples demonstrated by all of the Viking Biology Experiments. It is also proposed that the syntheses of these substances result mainly from electrical discharges between wind-mobilized particles at Martian pressures; plasmas are initiated and maintained by these discharges. Active species in the plasma either combine to form or react with inorganic surfaces to create the reactive constitutents.


Sarcoid of the nervous system.

Sarcoid is a granulomatous disease of undetermined etiology characterized by the presence of epithelioid cell aggregates without caseation which proceeds to conversion to hyaline fibrous tissue or resolution. The sites of nervous system involvement include the meninges, cranial, and peripheral nerves, hypothalamus and pituitary gland, muscles, and, more rarely, brain and spinal cord parenchyma. In nervous system involvement there is usually cerebrospinal fluid lymphocytic pleocytosis and elevated protein but these findings are not specific. When other systems are involved with sarcoid the diagnosis of neurosarcoidosis is obvious. When an unusual neurological symptom complex presents, sarcoid should be considered. The author reviews the literature and presents 22 cases of sarcoid involvement of the nervous system.


Experimental infection of wildebeest with the herpesvirus of infectious bovine rhinotracheitis/infectious pustular vulvovaginitis.

Intravaginal inoculation of a wildebeest (Connochaetes taurinus) with a wildebeest strain of the infectious bovine rhinotracheitis/infectious pustular vulvovaginitis herpesvirus induced only mild vulvovaginitis. The same virus did not produce any disease in another wildebeest exposed intranasally. A wildebeest bull which was inoculated by preputial instillation developed mild posthitis. The virus was reisolated only from the sites of inoculation. A carrier state was initiated in a wildebeest inoculated only once, intravaginally. The presence of this virus in the various secretions is a potential source for venereal transmission.


Powassan virus in Ixodes cookei and Mustelidae in New England.

Powassan virus was recovered from a pool of 3 nymphal and 1 adult female Ixodes cookei removed from a striped skunk (Mephitis mephitis) trapped in Massachusetts during 1967 and from a pool of 9 nymphal I. cookei from a long-tailed weasel (Mustela frenata) captured in Connecticut during 1978. Virus was detected in the blood of both mammals. Hemagglutination-inhibiting (HI) antibody to Powassan virus was demonstrated in 16.0% of the skunks sampled in Connecticut, and neutralizing antibody was detected in 83.3% of the skunks tested from Massachusetts. HI antibody was found in 1 of 6 long-tailed weasels from Connecticut and 1 of 6 short-tailed weasels (Mustela erminea) from Maine.


Morphologic evidence of poxvirus in "tattoo" lesions from captive bottlenosed dolphins.

Skin lesions known as "tattoos", were observed in several captive Atlantic bottlenosed dolphins (tursiops truncatus) maintained at the Naval Ocean Systems Center, San Diego, California. Biopsy of typical lesions and subsequent ultrastructural examination revealed intracytoplasmic particles with poxvirus morphology.


Pox infection among captive peacocks.

An outbreak of avian pox was detected among captive peacocks (Pavo cristatus) at Baghdad Zoological Park during spring, 1978. A total of 45 of the 60 birds in the aviary developed pox lesions around the beaks and eyes. Morbidity was 75% and mortality was 13%. A virus isolated from the skin lesions produced large plaques on the chorioallantoic membrane of developing chicken embryos and induced cytopathic effect characteristic for pox viruses in chicken embryo cell cultures. The virus neither haemagglutinated nor haemadsorbed to chicken erythrocytes. It was ether resistant and chloroform sensitive. Chickens inoculated with the virus by scarification developed localized pox-like lesions, while turkeys had only transient swelling of feather follicles at the site of inoculation. Virus partially purified with Genetron 113 was precipitated by antisera to fowlpox and pigeon pox viruses.


Secondary myocardial disease. Virus myocarditis and cardiomyopathy.

In an attempt to prove the hypothesis that virus myocarditis may be a cause of idiopathic cardiomyopathy, clinical and experimental studies were performed. Eleven patients with a presumptive or proven diagnosis of virus myocarditis were followed for one and a half to 13 years after the acute illness. One patient died in the acute stage, six recovered completely and one continued to have bifascicular block without subjective symptoms. Three patients had exertional dyspnea, cardiomegaly and an abnormal ECG three to 13 years after the onset, and two of them had an enlarged LV cavity with reduced EF and histological changes in myocardial biopsies. The clinical picture in these three cases was similar to that seen in congestive cardiomyopathy. Clinical observations of the heart in an epidemic of coxsackie B 3 virus infection among school children revealed that 49 (19%) of 263 infected children had abnormal chest-X ray, electrocardiographic or echocardiographic findings one to 10 months after the onset, however none of them developed cardiomyopathy. In experimental infection of weanling golden hamsters with coxsackie B 3 virus (Nancy strain), all animals developed acute and severe myocarditis, and the virus was detected in the myocardium. Hemodynamic data suggested decreased contractility of the left ventricle in the acute stage. Histologically the heart showed focal myocardial necrosis and cellular infiltration without calcification, findings which resemble those in human doxsackie B virus myocarditi. Thus, the golden hamster is a better animal model than the mouse in studies on virus myocarditis and postcarditic cardiomyopathy.


Experimental study of virus myocarditis in culture.

In this present paper, the authors would like to present here two points of results from the experimental study concerning virus myocarditis in culture. One is the direct delicate features of heart cells affected with Coxsackie B3 (Cox. B3) virus viewed through both light microscope and electron microscope. The other is the fact that marked proliferation of fibroblastic cells take place to replace the heart cells impaired by the virus suggesting to us the possibility of one of the genesisses, revealing fibroblastic involvement in the Idiopathic Cardiomyopathy (ICM) which could be followed by the residual of the virus myocarditis.


Effects of coronary vasodilator on cyclic nucleotides. The concentrations of cyclic AMP and cyclic GMP in canine coronary artery and left ventricular muscle following the administration of various coronary vasodilators.

We examined the effects of various coronary vasodilator drugs, papaverine, dipyridamole, isosorbide dinitrate, amyl nitrite, nitroglycerin, diltiazem, and nifedipine, on cyclic nucleotides of the coronary artery and left ventricular muscle of anesthetized dogs at maximum coronary blood flow after the administration of each agents. Only papaverine and dipyridamole significantly increased the concentration of c-AMP in the coronary artery. Nitroglycerin and siosorbide dinitrate did not significantly change the concentration of c-AMP but rather increased the concentration of c-GMP. Coronary vasodilator drugs were divided into three groups in association with the relationship of cyclic nucleotides in the coronary artery. Group I, including papaverine, dipyridamole, and amyl nitrite, increased the concentration of c-AMP and the ratio of c-AMP to c-GMP. Group II, including nitroglycerin and isosorbide dinitrate, incrased the concentration of c-GMP and decreased the ratio of c-AMP to c-GMP. Group III, including nifedipine and diltiazem, had no effect on the cyclic nucleotides. Group I drugs also increased the concentration of c-AMP in the left ventricular muscle and so group I drugs may predispose the ischemic heart to develop ventricular arrhythmias. It seems that the most useful coronary vasodilator is no effect on the c-AMP in the ventricular muscle and group II and group III drugs are more useful coronary vasodilator drugs than group I drugs.


Detection of erythrocyte membrane structural abnormalities in lecithin: cholesterol acyltransferase deficiency using a spin label approach.

The membrane fluidity of erythrocytes from patients with Lecithin: cholesterol acyltransferase (LCAT) deficiency was studied by means of electron spin resonance. The temperature dependence of the separation of the outer extrema of the spectra of 2-(3-carboxy-propyl)-4,4-dimethyl, 2-tridecyl-3-oxazolidinyloxyl spin probe was monitored for normal, presumed carrier and clinically affected subjects. The temperature profile of controls was significantly different from that of the presumed carriers and the clinically affected individuals. The results show that the compositional abnormalities previously noted in erythrocyte membranes from patients with LCAT deficiency are associated with alterations in the physiocochemical state of the membrane. An investigation of the spectral lineshapes below 10 degrees C allowed a distinction to be made at the membrane level between clinically affected subjects and clinically normal heterozygous carriers. Alterations in the temperature dependence of elec-ron spin resonance parameters may provide a sensitive index of red cell membrane alterations in pathological states of generalized membrane involvement.


Flow cytometry analysis of early DNA content changes in human and monkey cells following infection with Simian Virus 40.

Simian virus 40 (SV40) is capable of inducing cellular DNA synthesis in permissive and nonpermissive cells. Utilizing flow cytometry, we analyzed the DNA content changes in two diploid human cell strains and two monkey cell lines. The osteogenesis imperfecta (OI) human skin fibroblasts were induced into DNA synthesis, and within one to two cell generations, a polyploid cell population was produced. With WI-38 phase II cells, a similar pattern of increased cycling of cells into DNA synthesis was observed; however, the majority (approximately 60%) of the cells were blocked in the G2 + M phase of the cell cycle. At later time intervals, an increase in the G1 population was demonstrated. The two monkey cell lines responded to SV40 virus with an accumulation of cells in the G2 + M phase of the cell cycle. Thus, two diploid human cell strains exhibited different cell cycle kinetics early after infection with SV40 virus. The one strain (WI-38) behaved similarly to the two monkey cell lines studied. The other strain (OI) responded similarly to nonpermissive (transforming) cells infected with SV40 virus.


Endogenous cyclic AMP does not modulate transport of hexoses, nucleosides, or nucleobases in Chinese hamster ovary cells.

In a previous study we have demonstrated that neither extracellular nor intracellular cyclic adenosine monophosphate (AMP) levels directly affect the uptake of nucleosides, nucleobases, or hexoses by various types of cultured mammalian cells. Uptake of these nutrients into cells, however, involves two processes operating in tandem: facilitated transport across the membrane and intracellular phosphorylation; and uptake rates generally reflect the rates of substrate phosphorylation rather than of transport. In the present study we have examined the question of whether substrate transport per se is regulated by intracellular cyclic AMP. Initially various cell lines, grown both in suspension and monolayer culture, were screened for their cyclic AMP response to prostaglandin E1, isoproterenol, and inhibitors of cyclic AMP phosphodiesterase. Prostaglandin E1 treatment of Chinese hamster ovary cells was selected as the system giving the largest and most consistent (50-fold to 100-fold) elevation of cyclic AMP. Rapid kinetic techniques were used to measure the transport of 3-O-methylglucose, thymidine, adenosine, hypoxanthine, and adenine in wild-type cells and in mutant sublines incapable of phosphorylating these substrates. In no case was an increase in intracellular cyclic AMP accompanied by a significant change in the rate of transport of these substrates, although prostaglandin E1 slightly inhibited the transport of various substrates.


Colorectal carcinoma 15 years after the diagnosis of perianal Paget disease.

We report a patient with perianal Paget disease who developed a colorectal carcinoma fifteen years later, complicated by pseudomyxoma peritoneal. While perianal Paget disease is often associated with underlying carcinomas noted at the time of the diagnosis, we suggest that perianal Paget disease may also predispose to the future development of colorectal cancer.


Pontine gigantocellular field neuron activity time-locked with the PGO waves in the transitional phase of sleep in the cat.

The extracellularly recorded discharge of pontine gigantocellular tegmental field (FTG) neurons was studied in the cat during the transitional phase of sleep (TPS), which signifies in this study the shift from slow wave sleep (S) to paradoxical sleep (PS). The first appearance of pontogeniculo-occipital (PGO) waves, which were led off from the lateral geniculate nucleus, indicated the start, and the decrease of neck muscle tone the end of TPS. TPS was chosen because most PGO waves were then isolated and thus suitable for comparison with the discharge of neurons, which were localized to the antero-lateral part of the FTG. In order to analyze the possible role of these neurons in the control of PGO wave activity, the mean discharge rate of FTG neurons was studied in greater detail during 200 ms periods before and after the onset of each isolated PGO wave. Most FTG neurons discharged phasically and their activity was closely related to isolated PGO waves, so that the highest rate of unit discharge occurred within 40 ms period after the onset of the isolated PGO wave, being time-locked with its first deflection.


Urinary cyclic AMP and cyclic GMP in normal and asthmatic subjects during exercise.

The purpose of this study was to compare exercise-induced changes in urinary cyclic AMP and cyclic GMP of normal and asthmatic subjects. All subjects were exercised to 80% of predicted maximum heart rate for 15 minutes on a bicycle ergometer. FEV1 was measured before and after exercise to determine the degree of bronchoconstriction due to exercise. Five urine specimens were collected by voluntary voiding at 20 minute intervals (one specimen before exercise and four after exercise). Before exercise, the asthmatic subjects excreted less cyclic GMP (p less than 0.05) but the same cyclic AMP as the normal subjects. Creatinine excretion in nine asthmatic subjects decreased by 24% (p less than 0.01) during exercise. In sixteen normal subjects (post-exercise bronchoconstriction less than 10%) cyclic AMP excretion increased by 19% (p less than 0.05) within 20 minutes after exercise, while cyclic GMP excretion did not change significantly. In thirteen asthmatic subjects (post-exercise bronchoconstriction greater than 20%) cyclic AMP excretion did not change significantly, while cyclic GMP excretion increased by 23% (p less than 0.05) within 20 minutes after exercise. This study has shown that exercise affects the excretion of cyclic AMP, cyclic GMP, and creatinine in normal and asthmatic subjects. The response of normal and asthmatic groups to exercise was the same for creatinine but different for cyclic AMP and cyclic GMP.


Involvement of cyclic AMP and its receptor protein in the sensitivity of Escherichia coli K 12 toward serine: excretion of 2-ketobutyrate, a precursor of isoleucine.

A relationship between serine-induced growth sensitivity and the cAMP-CAP complex is established. Mutants of Escherichia coli K 12 deficient either in the cya or crp gene function exhibit a resistant phenotype on serine media although they harbor a relA allele normally leading to sensitivity toward serine. The presence of a crp allele in a cya delta relA background restores the sensitivity phenotype, while the analysis of serine resistant mutants selected from a crp cya delta relA strain shows that the mutation leading to resistance is located at, or very near, the crp gene, giving a more or less Crp- phenotype. In addition crp cya delta relA strains excrete large quantities of 2-ketobutyrate when grown on glucose M63 medium. This excretion is unambiguously linked to the presence of the crp allele and is correlated with an enhanced threonine deaminase activity. Besides, the complex regulation exerted on the acetolactate synthase activities is discussed.


The stimulation of Escherichia coli stringent factor-dependent synthesis of guanosine 3',5'-polyphosphate [(p)ppGpp] by rat liver ribosomal proteins.

The effect of groups of proteins from rat liver ribosomes on the Escherichia coli stringent factor-catalyzed synthesis of (p)ppGpp was tested. Most groups were capable of supporting (p)ppGpp synthesis; the exceptions were A40, B140, B240 and B160 which contain proteins which are relatively less basic than those in the active groups. The capacity of 30 individual rat liver ribosomal proteins to activate stringent factor was assessed; most sustained the synthesis of (p)ppGpp. Proteins S12, S21, L12, P1, and P2 (which are acidic or relatively acid) had no activity; proteins S6, S8, and L3 were the most active: the others had moderate activity.


Control of minicell producing cell division by cAMP-receptor protein complex in Escherichia coli.

It has been established that the strain CA8000 of Escherichia coli K12 produces minicells. This phenotype of CA8000 has been shown to be suppressed by additional mutations in cya or crp genes. Minicell production by cya+ crp+ min bacteria is probably a consequence of error, introduced by horizontal growth, in the selection of site on the envelope for initiation of hemispherical growth.


A possible role of estrogens in carcinogenesis of non-target tissues.

The mitogenic action of the estrogen-receptor complex is supposedly similar in both normal and malignant target tissues. As receptors are present in several types of non-target tissues, in the case of lesions at the nuclear acceptor sites, the complex in those might be able to cause successive mitoses. Estrogen-dependent tumors of non-target tissues have been reported by several investigators. In normal and malignant cells of the breast and some other types of non-endocrine cells, the ability to produce their own estrogens (from circulating precursors) has been shown. The locally formed estrogens might have a role in the initiation of some malignant transformations. Indications of this process are the switching to estrogen production of some neoplastic endocrine or undifferentiated cells, certain ectopic effects displayed by some cancerous tissues, and the possible roles of GH, PRL and cholesterol in the development of some malignancies. The present endocrine system for the synthesis of the sexual hormones might be a specialization of a system at the cellular level. Polypeptide hormones might evolve from regulatory parts of cyclases or phosphodiesterases. Traces of the original biological processes might still be maintained by several cell-types.


Metabolic effects of oral contraceptives containing 30 micrograms and 50 micrograms of oestrogen.

In Study A, biochemical data from 17 women who were not taking oral contraceptives were compared with those from women taking preparations which contained either 30 microgram of ethinyl oestradiol and 150 microgram of D-norgestrel (18 women) or 50 microgram of ethinyl oestradiol and 250 microgram of D-norgestrel (nine women). In Study B, biochemical data were collected from eight women before and during the first three or four months therapy with preparations containing 30 microgram of ethinyl oestradiol and 150 microgram of D-norgestrel. The two oral contraceptive dosage forms studied produced qualitatively and quantitatively similar metabolic changes. Both caused an increase in serum concentration of triglycerides (30% to 33%), beta-lipoproteins (27% to 29%) and ceruloplasmin (75% to 90%), and a decrease in serum levels of antithrombin III (22% to 29%) and ascorbic acid (30% to 42%). Serum cholesterol and phospholipid concentrations were unchanged. However, the proportion of serum cholesterol carried by alpha-lipoproteins (high density lipoproteins) decreased, while that carried by beta-lipoproteins (low density and very low density lipoproteins) increased. The former change is in the same direction, but much smaller than that observed in coronary heart disease.


[The problems of lipid metabolism. Demands for diagnostically improved insight into the function of lipoproteins (author's transl)].

In the course of a follow-up of 87 subjects with known hyperlipoproteinemia a new, simplified method was tested for quantification of the individual lipoprotein fractions capable of being performed by every larger laboratory, in contrast to the expensive ultracentrifugation. The LDL/HDL ratio proved to be the conclusive parameter for daily diagnosis in hospital and general practise. This enabled the most important aspects of the lipid metabolic situation of a patient to be recognized immediately.


Effects of dipyridamole in vivo on ATP and cAMP content in platelets and arterial walls and on atherosclerotic plaque formation.

In rabbits receiving an atherogenic diet for 2 months, the ATP content of platelet rich plasma (PRP) and arterial tissue was significantly elevated as compared to normal rabbits. This increase in ATP levels of platelets from atherosclerotic rabbits was paralleled by higher basal as well as PGI2-induced cAMP levels. In arterial tissues, an increase was only obtained in PGI2-stimulated cAMP content. Treatment with dipyridamole (DPD) for 4 weeks resulted in a reduction of the ATP content in platelets and arterial tissue from atherosclerotic rabbits to values seen in normal animals. Again, the reduction of ATP content was reflected in a decrease of basal as well as PGI2-induced cAMP levels in platelets, whereas in arterial tissue a decrease was only obtained in PGI2-induced cAMP content. At the same time, DPD treatment enhanced atherosclerotic plaque formation in the aortic wall. The enhanced atherosclerotic plaque formation seen in DPD treated atherosclerotic rabbits may be linked to the inhibition of adenosine uptake, resulting in a decrease of the adenine nucleotide pools of arterial wa-l cells. The decrease also caused a reduction in PGI2-induced cAMP content. This effect may be linked to altered proliferative activity, since in many cell types, stimulation of cAMP levels results in reduced proliferation rates.


Interaction of diazepam with synaptic transmission in the in vitro rat hippocampus.

Diazepam (5 x 10(-8) - 10(-6) M) was found to augment recurrent inhibition of pyramidal neuron firing in a dose-dependent manner in rat hippocampal slices. To determine possible loci of this effect, diazepam was locally administered by pressure ejection from a micropipette, while recording action potentials from single inhibitory ("basket") interneurons. Diazepam induced reversible and reliable increases in interneuron firing in response to stimulation of Schaffer collateral and commissural afferents. Taken together with previous electrophysiological reports, these data suggest that benzodiazepines may augment central inhibition by increasing either the excitability of inhibitory interneurons, or by increasing the strength of excitatory afferents to these cells.


Increased susceptibility of leukemia-infected chickens to Marek's disease.

Intramuscular (i.m.) injection of RAV-49 into 1--3 day-old chickens infected with Marek's disease herpes virus (MDHV) by contact led to exceedingly high rate of Marek's disease (MD) incidence. According to the data obtained in three experiments 52 out of 108 (48.1%) chickens fell ill in this group. While in the group uninfected with RAV-49 but contactly infected with MDV-Kekava there were only 11 out of 102, (10.7%) incidences. Simultaneous inoculations of RAV-49 and MDV had no effect on MD morbidity. Increased susceptibility of i.m. RAV-49-infected and contactly MDV-infected chickens to MD was abolished by simultaneous inoculation of RAV-49 and herpes virus of turkeys (HVT).


Studies of the leukemic cell associated antigen(s) in mice with myeloid leukemia (Graffi), using the immunoperoxidase techniques.

Thymus, spleen and lymph nodes from myeloid leukemic mice (Graffi) were examined by light and electron microscope using immunoperoxidase techniques. As immune sera autologous sera were applied. The percentage of the labeled leukemic cells ranged from 30 to 60. It was established that leukemic cell associated antigen(s) was located mainly on the plasma and nuclear membranes. Occasionally this antigen(s) was observed in some areas of the endoplasmic reticulum, on separate cytoplasmic vesicles, on the budding viruses and on the chromatin. In addition, weakly expressed antigen(s) was located in cytosol. The mature virus particles were not labeled with peroxidase.


Conditions for hormone-stimulated expression of endogenous C57Bl strain-associated mammary tumor virus genome.

Full MMTV-Y gene expression can occur in dexamethasone-insulin-prolactin-stimulated cell cultured derived from C57Bl/10 mammary adenocarcinomas induced by syncarcinogenic action of chemical carcinogens (dimethylbenzanthracene), and mammotropic hormones (estrogen and prolactin). The rate of the hormone-stimulated virus production, as determined by biochemical, immunological and electron microscopical methods, was comparable to the level of spontaneous MMTV production by cells of established mouse mammary cancer cultures (CCL-51 and Mm5mt/cl). On the contrary, no virus production has been detected in hormone-stimulated cultures derived from C57Bl/10 mammary tumors induced by chemical carcinogen alone (urethan).


Carcinoembryonic antigen (CEA) in patients with malignant and non-malignant diseases.

Serum carcinoembryonic antigen (CEA) concentration was found to be raised in 503 of 550 patients (91%) with bladder cancer, lymphoma of intestine, hepatocellular carcinoma, bronchogenic carcinoma, prostate cancer, cirrhosis of liver and bilharziasis. The degree of elevation was moderate in all patients except in 189 patients in whom values more than 20 ng/ml were recorded, of which 53 patients with bladder cancer and 118 patients with bilharziasis. The mean CEA value in the patients with cirrhosis in the non-tumorous liver was slightly higher than that in those without cirrhosis, but the difference did not reach statistical significance (P greater than 0.01). There was no correlation between serum CEA and alph-fetoprotein (AFP) levels in all patients except in patients with bladder carcinoma, hepatoma and bilharziasis.


Compared localizations of prolactin-like and adrenocorticotropin immunoreactivities within the brain of the rat.

By an immunoperoxidase (PAP) technique, ACTH- and prolactin-like substances were detected in neurons of several hypothalamic nuclei in young and adult rats. Neurons in the arcuate and ventromedial nuclei were simultaneously both ACTH-like and PRL-like positive. Labelled fibres were followed to their fields of projections. Intensity of labelling was not influenced by experimental procedures known to change PRL and ACTH content of the pituitary. Colchicine enhanced labelling of perikarya while lowering that of fibres. The results support neuronal synthesis of hormonal ACTH and PRL related molecules and their involvement in the control of autonomic nerve functions.


Nelson's syndrome: one disease or two?

We present a case of Nelson's syndrome in which the clinical, radiographic, and surgical findings are indicative of a discrete microadenoma rather than of diffuse pituitary hyperplasia. Ultrastructural analysis reveals this to be the first example of a sparsely granulated lesion in Nelson's syndrome and only the second such adrenocorticotropic hormone-secreting tumor reported. Unfortunately, there do not seem to be any firm ultrastructural features that differentiate between hyperplasia and discrete adenoma. Separation of the two entities may be important in delineating the pathogenesis and treatment of Nelson's syndrome; these are discussed.


Serum cortisol abnormalities after craniocerebral trauma.

Serial estimations of serum cortisol were performed in 49 patients with craniocerebral trauma. Abnormalities of serum cortisol, including alterations in diurnal rhythm and elevations of serum cortisol level, occurred in 21 patients. The frequency and severity of the abnormalities correlated with the severity of the head injury, and there was a trend suggesting that middle fossa basal skull fractures predisposed to cortisol abnormalities. A further 6 patients were studied to assess the effects of exogenous dexamethasone, and in all patients there was suppression of elevated serum cortisol levels by the dexamethasone. The findings suggest that hypercortisolemia after head injury is related to an alteration rather than an abolition of the normal feedback mechanism. (Neurosurgery, 5: 559--565, 1979).


Use of metrizamide computerized tomographic cisternography in the evaluation of patients with malignant glioma for immunotherapy.

The communication between the subarachnoid space and the surgically created tumor cavity in glioma patients was evaluated by metrizamide cisternography. It was necessary to know the presence or absence of such a communication to determine the route of administration of autologous lymphocytes as a form of immunotherapy. The contrast injections were made either into the spinal subarachnoid space or directly into the tumor cavity. The presence of communication was demonstrated and followed by computerized tomographic (CT) scanning. The results were also corroborated by comparing the white cell counts in the fluid from the tumor cavity and the spinal subarachnoid space 24 hours after autologous lymphoid cell infusion. In only two of seven patients was a communication present. In the five patients without a communication, the blocks were at the tentorial hiatus (one patient), due to a nonpatent subarachnoid space over the cerebral convexity (two patients), and the result of adhesions at the pial margin of the tumor cavity (three patients). In addition, certain limitations in the use of computerized tomography in the evaluation of glioma patients are demonstrated. These problems include the effects of steroids on tumor size, the poor correlation between "enhancement" on CT scan and tumor recurrence, and the difficulty of differentiating metrizamide and hemorrhage by CT scan in the immediate postoperative period. (Neurosurgery, 5: 576--582, 1979).


[Considerations on a series of malaria cases].

Stress is laid on the seriousness of malaria and the causes of its unusual clinical pictures. Results obtained in the evaluation of G-6-PD, PK, intracellular K and globular resistance, in addition to other parameters, suggest (as pointed out in the literature) that altered cell metabolism may be the primary cause of haemolysis. Attention is also drawn to the lengthy persistance of gametes after gametocidal management in some patients, and the risks that this involves.


The role of coxsackie B viruses in heart disease in New Zealand.

Sera from 622 patients received in a 20-month period were examined for coxsackie B neutralising antibody to serotypes 1--6. Two hundred and thirty-four of the total 622 were patients with a cardiac symptomatology, and 388 formed a comparable group of non-cardiac cases. There were 141 (60 percent) seropositive patients in the cardiac group, 34 percent and 29 percent of which respectively had antibody to B4 and B2. The clinical data provided on these 141 patients indicated that 33 (23 percent) had acute myocarditis, and 59 (42 percent) had acute pericarditis; 41 (29 percent) were recorded as having acute pleurodynia or Bronholm disease while eight (6 percent) had other indications of cardiac disease. Amongst the cardiac patients the highest prevalence of seropositivity was found in the 20--39 age group and 67 percent of these were males.


[5-fluorouracil inhibits bacterial collagenase and the collagenolytic system of a human rhabdomyosarcoma].

Protease inhibitory activity of eight cytostatic drugs on 5 proteases was tested by applying an immunoelectrophoretic system, a rhabdomyosarcoma's collagenolytic activity was investigated and inhibition studies were performed. We found 5-fluorouracil stopping collagenolytic activity of clostridial origin and the enzyme released by the tumor. No other cytostatic drug showed protease inhibition. The collagenolysis of the rhabdomyosarcoma was examined for its origin. Negative inhibition studies with ethylene-diamine-tetraacetate make the leukocyte origin highly improbable so that one can suggest that the hydrolytic enzyme derived from the tumor. Our findings could contribute to characterize the enzyme released by the tumor. We  suggest its role as a mediator of invasion and metastasis and on the other hand we detected its relatively specific inhibitor, 5-fluorouracil, which could influence tumorous and normal growth, regeneration and the intermediary protein metabolism.


[Pressing problems of after care of patients with cancers of the cervix and corpus uteri].

A modern and optimal follow-up treatment of patients with uterine cancer requires a detailed knowledge of this disease and its sequelae. Incidence and localisation of cervical and endometrial cancers as well as kind and incidence of therapeutical side effects are demonstrated in the cases of the Gynecological University Hospital of Freiburg, GFR. A study of the psychosexual situation 2--4 years after therapy of uterine cancer is reported. Actual state of social assurances for cancer patients in the German Federal Republic is pointed out. The dates allow to draw conclusions for the follow-up of individual cases.


[Familial Hodgkin's disease: a case report and review of the literature].

In a family of five sisters four developed malignant lymphomas. A case report is given together with a literature survey of familial Hodgkin's disease. The observations fit the hypothesis that for the development of Hodgkin's disease two requirements must be met: a genetically determined susceptibility as well as some causative agent.


[Enzyme induction by Epstein-Barr-virus-producing infection of lymphoblastoid cell lines].

By means of biological in vitro assay collagenolytic activity of normal peripheral lymphocytes was compared to that of EBV-infected lymphoblastoid cell lines: the EBV-producing P3HR1 cells and the non-productive Raji cells. While normal lymphocytes and Raji cells revealed only traces of activity, significantly increased (p less than 0.001) collagenolytic activity was detected in samples of P3HR1, the cell-free EBV-suspension alone showed no collagenolysis. The described enzymatic activity was fully inhibited by 0-phenanthroline and EDTA indicating collagenase action. As main explanations for the phenomenon we suggest the possibility of viral enzyme induction, increased release of protease and thirdly that EBV carries a gene for synthesis of a collagenase.


[Cyclic nucleotide concentration changes in different tumors and therapeutic success through increasing the cAMP level].

Changes in the concentration of cyclic AMP as well as cyclic GMP were measured in different murine tumors and in human tumors of varying malignancy. The quotient of cAMP and cGMP seems to be an important parameter for the molecular-biological derangement. Because of the recently much discussed importance of cAMP and cGMP in the immune defence the changes in the concentration of both nucleotides were measured in the T-lymphocytes of tumor patients. Significant changes occurred in patients with malignant melanoma. Investigations of the stimulatibility of the cAMP and cGMP levels revealed a diminished activatibility of the cAMP level and a higher stimulatibility of the cGMP level in the T-lymphocytes of patients with malignant melanoma as compared with those of the controls. On the basis of the working hypothesis that there is a causal relationship between the deranged dualism of cAMP and cGMP in the T-lymphocytes and the failure of the immunological tumor cell defence, an increase in the cAMP level is offered as a possible therapy. Therapeutic results in tumor-bearing mice and first results in melanoma patients are discussed.


Cranial nerve damage in patients after alpha (heavy)-particle radiation to the pituitary.

The records of 161 patients were reviewed to determine if radiation damage had occurred following cranial irradiation. All of these patients had received alpha-particle radiation to their pituitary glands during the period when this form of therapy was given for diabetic retinopathy. Extraocular muscle palsy developed in 11 of these patients, iridoplegia in six, and fifth nerve damage in six. All of the palsies developed within a short period following their irradiation, and a definite dose relationship was present. The dose rate was approximately 100 rads/min for all cases. Fractionation varied but it is known for all cases. The estimated doses to the third, fourth, fifth, and sixth cranial nerves was calculated at a saggital plane 13 to 15 mm from the pituitary by using computer-drawn dosimetry charts for the respective aperture size. The energetic alpha particles were produced by the 184-in synchrocyclotron at Berkeley, Calif. A dose relationship for radiation palsies was apparent.


Aniridia caused by a heritable chromosome 11 deletion.

A child with aniridia, multiple anomalies, severe failure to thrive, and severe psychomotor retardation is shown to have a syndrome similar to, though more severe than, other patients with overlapping deletions of the short arm of chromosome 11 (Pediatrics 64:604, 1978). Her deletion (46,XX,del [11p] [pter yields p14::p11.3 yields qter]) was derived from her mother, who has a chromosome 11 shift (46,XX,der [11] [pter yields p14::p11.3 yields q22::p14 yields p11.3::q22 yields qter]). The significance of del (11p) in the aniridia-Wilms' tumor association is discussed, and the del (11p) basis for aniridia is compared with other genetic bases for aniridia.


[Human erythrocyte protein kinases (author's transl)].

Cyclic AMP dependent and independent protein kinase activities are present in membrane as well as cytosol of human erythrocyte. Red cell membrane contains at least: 1) a cAMP dependent histone kinase type I using ATP as phosphoryl donor; 2) a cAMP independent casein kinase using ATP or GTP as phosphoryl donor; this kinase probably phosphorylates erythrocyte spectrin. In lysate of human red cell there are two or three cAMP dependent protein kinase type I and II and one cAMP independent casein kinase. Characteristics of these protein kinase activities are reviewed. Membrane substrates are numbered taking in account that the activity assays are often of dubious value. Several phosphorylation abnormalities previously reported in hematologic and non-hematologic disorders are still uncertain and at present time not any red cell protein kinase deficiency has been demonstrated.


An analysis of the peritonitis in acute murine reovirus infection.

Reovirus 3 replicates in the mesothelial cells of the peritoneal cavity of infected neonatal Prince Henry mice. As virus replication becomes evident in these cells lymphocytes, polymorphonuclear leucocytes and especially mononuclear phagocytes increase in number in the peritoneal exudate and often surround the exfoliated and degenerating infected mesothelial cells. These events are accompanied by a polymorphonuclear leucocytosis, a lymphopenia and a transient monocytosis which reverses to a monocytopenia as virus replication appears in a variety of organs. Although in the peritoneal cavity the majority of exuded monocytes ingest virions and cellular debris a small proportion supports replication of reovirus 3. Polymorphonuclear leucocytes, on the other hand, did not exhibit evidence of reovirus replication.


The pathological assessment of ovarian neoplasms. III: The malignant "epithelial" tumours.

One thousand common "epithelial" tumours of the ovary were encountered in a 25-yr study period at the King George V Memorial Hospital. These tumours were classified according to the World Health Organisation (W.H.O.) Histological Classification of Ovarian Tumours. In this report a detailed histological assessment of the 298 malignant "epithelial" tumours is presented, including criteria for histological grading and typing. Evidence for the multifocal tumorigenesis of serous tumours and the pathological correlates of endometrioid carcinoma are stressed.


Influence of aminophylline and cyclic AMP on glycogen metabolism in fetal rat lung in organ culture.

The glycogen content of fetal rat lung declines coincident with increased pulmonary phospholipid synthesis. Aminophylline, a methylxanthine cyclic adenosine 3',5' monophosphate (AMP) phosphodiesterase inhibitor, and cyclic AMP augment fetal lung phospholipid synthesis. Because lung glycogen breakdown may contribute to pulmonary phospholipid synthesis, the effects of aminophylline and cyclic AMP on glycogen metabolism were studied in explants of 19 day fetal rat lung in organ culture. Treatment with aminophylline or dibutyryl cyclic AMP for 24 hr, resulted in a 25% (P less than 0.025) and 75% (P less than 0.001) decrease, respectively, in the glycogen content of the explants. Glycogen synthase I activity was reduced by 32% in aminophylline treated cultures (P less than 0.025) and 25% in cyclic AMP treated cultures (P less than 0.025). The percent of total synthase in the active form was significantly reduced in all treated cultures. Neither aminophylline nor cyclic AMP treatment resulted in significant changes in glycogen phosphorylase a or total phosphorylase activity.


Hormonal phenotype and HLA-genotype in families of patients with congenital adrenal hyperplasia (21-hydroxylase deficiency).

The response of 17-hydroxyprogesterone (17-OHP) and cortisol (F) to a 6-hr ACTH stimulation in families of patients with congenital adrenal hyperplasia due to 21-hydroxylase deficiency was studied. These studies demonstrated that siblings who should be heterozygous carriers of the 21-hydroxylase deficiency gene based on HLA genotyping are hormonally different from the general population. In pre- and early pubertal children predicted to be heterozygous carriers of the gene based on HLA genotyping, the 17-OHP level (13.1 +/- 4.5 ng/ml), the rate of increase of 17-OHP (0.03 +/- 0.01), and the ratio of 17-OHP/F at 6 hr (0.27 +/- 0.07) were significantly higher (P less than 0.001) than in the control population, (3.9 +/- 1.9, 0.009 +/- 0.005, and 0.08 +/- 0.04 ng/ml, respectively). In late and postpubertal males, these hormonal parameters in the heterozygotes (17 +/- 9.7, 0.04 +/- 0.026, 0.42 +/- 0.33 ng/ml, respectively) were significantly higher (P less than 0.001) than in the general population (5.3 +/- 1.6, 0.009 +/- 0.004, and 0.1 +/- 0.03 ng/ml, respectively). In postmenarchal females, the mean hormone responses in the heterozygotes (12.1 +/- 9.7, 0.03 +/- 0.02, and 0.27 +/- 0.24 ng/ml, respectively) were significantly higher (P less than 0.005, less than 0.01, less than 0.005, respectively) than in the general population (5.2 +/- 2.5, 0.01 +/- 0.007, and 0.1 +/- 0.04 ng/ml, respectively). However, the overlapping values did not permit a clear differentiation of the hormonal responses in these two groups. Another (ACTH) stimulation in one family demonstrated that a father of a patient probably is a previously unrecognized homozygous affected patient and,thus, revision of the congenital adrenal hyperplasia (CAH) genotype for this family was required.


Deficiency of glucose-6-phosphate dehydrogenase found in a case of hepatic fructose-1,6-diphosphatase deficiency.

The first case of fructose-1,6-diphosphatase (FDPase) deficiency in Japan showed a decreased activity of glucose-6-phosphate dehydrogenase (G6PD) in the liver, white, and red blood cells. In the enzymatic study of G6PD which was partially purified from red cells, the following characteristics were observed in the enzyme of the patient. 1) The G6PD activity of the patient was reduced to 17% of normal, but no evidence of a hemolytic episode was found in his past and family history. 2) In the investigation of G6PD of the patient, no abnormalities were observed in its enzymatic parameters such as electrophoretic mobility, Km for G6P and NADP, Ki for NADPH, the utilization of 2-deoxy G6P and deamino NADP, heat-stability, and pH curves. 3) The dissociation constants of red blood cell G6PD for NADP and NADPH, which were obtained from the investigations on the reactivation of cold-inactivated G6PD at 37 degrees C, were about 3 times higher in the patient as compared to the values of the normal controls. Based on these findings, it might be concluded that the G6PD deficiency found in the red blood cells of this case of a FDPase deficiency is a unique variant, which could not be characterized by using only the method recommended by a World Health Organization (WHO) scientific group. Considering that the abnormality observed in the G6PD of this patient was a decrease in the affinity of the enzyme for its coenzymes, the dissociation constants for the coenzymes in reactivation process might be another important kinetic parameter in characterizing the G6PD deficiency.


Role of the basal sodium intake in the rats on their response to a natriuretic factor.

A previously described natriuretic factor found in urine from man and dogs receiving a high salt diet has been postulated to be of hormonal nature. In order to test this hypothesis, natriuretic factor obtained from a pool of human urine of a standard activity has been injected into rats fed 0.55 (group A), 3.55 (group B), 6.55 (group C) mEq sodium a day as it has been shown that the endogenous activity of this factor depends on the sodium content of the diet. This material induces an increment of the UNaV and FENa in the rats averaging respectively 429.6 +/- 150.1% and 317.5 +/- 71.14% for the rats from group A, 103 +/- 29% and 99.7 +/- 26.25% for the rats from group B, 56.3 +/- 34.4% and 63.2 +/- 35.8% for the rats from group C reflecting the degree of statistical significance observed between the absolute values obtained during the control and experimental periods for each group. In addition, a dose-response curve has been established. Altogether the results suggest the hormonal nature of the natriuretic factor and design the kidney as the target organ. These data allow to define the best conditions under which the rats have to be in order to provide a valuable bioassay for this natriuretic material.


Serotoninergic mechanisms of beta-endorphin-induced hypothermia in rats.

The effects of intraventricular administration of beta-endorphin on thermoregulatory responses of unanesthetized rats to different ambient temperatures (Ta) of 8, 22 and 30 degrees C were assessed. Administration of beta-endorphin produced a fall in rectal temperature at both Ta 8 and 22 degrees C. The hypothermia in response to beta-endorphin was brought about by both cutaneous vasodilation (as indicated by an increase in both the tail and the foot skin temperatures) and decreases in metabolic heat production. However, at Ta 30 degrees C, administration of beta-endorphin produced no change in rectal temperature or other thermoregulatory responses. Furthermore, the hypothermic effect induced by beta-endorphin was greatly attenuated by either the depletion of brain serotonin levels (with 5,6-dihydroxytryptamine and p-chlorophenylanine) or the blockade of opiate receptors (with naloxone). The data indicate that beta-endorphin leads to hypothermia in rats by increasing sensible heat loss and decreasing metabolic heat production, probably via the release of endogenous serotonin within brain.


Description of the firing pattern of respiratory neurons by frequency modulated interspike interval distributions.

The description of the firing pattern of respiratory neurons presented here is based on extracellular recordings and directed to the possible influence (facilitating of inhibiting effect) that the neuron might have upon others (output) as well as to the extend of influences that are received from other neurons (input). Developing such a description the spike generating mechanism of the neuron was in a first approach characterized by a frequency modulated gamma distribution. Using this approach an acceptable match between calculated and measured interspike-interval distribution was found although mostly not statistically significant. This non-significancy is most probably due to the choice of the simple gamma distribution as a first approach. However, applying this description it seems possible to retrieve statistical information about the interaction between neurons from extracellular recordings. Based on this description it is proposed to use modal spike frequency as well as the variance of the interspike interval distribution as estimates for the activity and the extent of activation of the neuron.


The role of the adrenergic innervation to the pancreatic islets in the control of insulin release during exercise in man.

The normal depression of plasma insulin concentration during exercise has been ascribed to adrenergic inhibition of insulin release and the role of humoral catecholamines in this hormonal adjustment has repeatedly been stressed. In the present study this contention has been investigated in 6 bilaterally adrenalectomized patients and in 6 sex- and age-matched controls who undertook exercise on an ergometer until they were exhausted. No differences were observed in any cardiovascular or metabolic adjustments between the two groups during strenous exercise. Mean plasma insulin concentration fell by about 50% in both groups. Phentolamine effectively abolished the fall in plasma insulin concentration during exercise in 2 adrenalectomized patients. The results suggest that the adrenergic nerves that supply the B-cells have a functional role in man during exercise.


The steady state response of brainstem respiratory neuron activity to various levels of PA,CO2 and PA,O2.

Extracellular recordings were made of 52 respiratory neurons in the brainstem of cats, anesthetized (chloralose-urethane), vagotomized and artificially ventilated. Phrenic nerve activity was recorded and quantified as an index of the output of the respiratory neuronal organization in the brainstem. The unit activity was quantified by using the modal spike frequency as a possible indication of the activating effect of one unit on other respiratory neurons (Smolders and Folgering, 1979). Inspiratory neurons showed the strongest reaction to changes in PA,CO2 and/or PA,O2. Expiratory neurons and frequency modulated neurons responded less to changes in chemical drive. Phase spanning neurons did not show any consistent response. Four out of ten continuously firing neurons without any respiratory rhytmicity increased their firing frequency when PA,CO2 was increased. Apart from the increase in modal spike frequency, the respiratory neuronal organization also reacted with an increase in active units (recruitment) when the chemical drive was increased. The relationship between quantified phrenic nerve activity and spike frequency was independent of the stimulus (hypercapnia or hypoxia). A model was developed in which the increase in modal frequency of a unit arouses other units: when the chemical drive increases, progressively more units tend to be recruited into the respiratory neuronal organization in the brainstem.


Purification of the mRNA for chicken very low density lipoproteinII and molecular cloning of its full-length double-stranded cDNA.

The mRNA coding for the small apo-Very Low Density Lipoprotein (apo-VLDLII) from chicken serum was highly enriched by oligo(dT) chromatography and preparative gel electrophoresis of estrogenised liver RNA. Double-stranded cDNA was synthesised by the subsequent actions of reverse transcriptase and DNA polymerase, and used for a preliminary characterisation of the structural gene. Molecular cloning of dC-tailed ds-cDNA into the Pst I site of plasmid pBR 322 yielded several recombinant clones. Five chimeric DNAs were selected and characterised by restriction enzyme mapping and electron microscopy of R-loops. At least two of them (pVLDLII 3.33 and pVLDLII 4.82) contain an almost full-length ds-transcript of VLDLII mRNA in which no more than 10-20 bases at the 5'- end are missing.


Synthesis of the human insulin gene. Part II. Further improvements in the modified phosphotriester method and the synthesis of seventeen deoxyribooligonucleotide fragments constituting human insulin chains B and mini-CDNA.

The purification of protected deoxyribooligonucleotides containing phosphotriester internucleotidic linkages has been improved by developing a deactivated silica gel chromatographic technique. The efficiency of this technique as applied in the modified phosphotriester approach has been demonstrated in the rapid synthesis of seventeen pure fragments constituting the sequence of human insulin B and mini-C DNA. The sequence of each oligomer was confirmed by the two-dimensional mobility shift method of fingerprinting.


Regions of the polyoma genome coding for T antigens.

The early region of the polyoma genome encodes three T antigens. We have analyzed the organization of the coding regions for the T antigens, using the nucleotide sequence of polyoma DNA and peptides derived from purified, radio-labeled T antigens, separated by two-dimensional electrophoresis and chromatography. We compared the peptides, predicted from the nucleotide sequence of the DNA, with those derived from the purified T antigens. We also compared chemically synthesized peptides, predicted from the DNA sequence, with observed peptides. The results show that the three polyoma T antigens are encoded in overlapping regions of the viral DNA, translated, in part, in two different reading frames.


Experimental evidence for asymmetrical shielding of nucleosomal DNA by histones.

Electron spin resonance study of Mn (II) binding to chromatin and derivatives, including core particles, shows that Mn (II) is a good probe for testing the overall electrostatic balance of the nucleoproteic complex as well as DNA accessibility. Experimental results are in good agreement with a recent model proposed (Mirzabekov A. D. and Rich A. (1979) Proc. Natl. Acad. Sci. USA 76, 1118-1121), for the core particle, in which an asymmetrical shielding of DNA by the protein core is assumed. Furthermore, it was found that the histone H1 hinders a number of charges on the linker DNA in a proportion equal to the net positive charge of the histone itself. This result is interpreted as due to a tighter interaction between the linker DNA and the core histones in the presence of histone H1.


Intracellular distribution of hepatic and renal gluconeogenic enzymes in embryonic and growing chickens.

The intracellular distribution of hepatic and renal gluconeogenic enzymes in 20-day-old chicken embryos and 4-week-old chickens (Gallus domesticus: New Hampshire male X Columbian female) has been studied. Pyruvate carboxylase, fructose-1,6-diphosphatase, and glucose-6-phosphatase were found primarily in the mitochondrial, cytosolic, and microsomal fractions, respectively. Phosphenolpyruvate carboxykinase was present not only in the mitochondria but also in the cytosol of the chicken liver and the kidney. The intracellular distribution of the liver enzyme differed from that of the kidney enzyme in chicken embryos as well as in growing chickens.


The cholinergic system of the brain and its interactions with other aminergic systems.

The new achievements in the understanding of acetylcholine (ACh) synthesis, storage, metabolism, the neuronal action, the structure of cholinergic synapses and the nature of cholinergic receptors in the central nervous system are described. Moreover, interactions of cholinergic mechanisms with adrenergic, serotoninergic, GABA-ergic and histaminergic systems in the brain are discussed.


Implantation of the isolated human erythrocyte anion channel into plasma membranes of Friend erythroleukemic cells by use of Sendai virus envelopes.

Sendai virus envelopes and human erythrocyte band 3 membrane polypeptides were isolated in detergent solutions and coreconstituted into detergent-free vesicle structurally resembling viral envelopes. Formation of hybrid viral envelope-band 3 vesicles (VB3) was demonstrated by immunoprecipitation of VB3 containing 4,4'-diisothiocyano-2,2'-ditritostilbenedisulfonic acid-labeled band 3 in the presence of antiviral antiserum. The hybrid VB3 vesicles displayed agglutination, fusion, and hemolytic properties similar to those of reconstituted viral envelopes. Incubation of VB3 with Friend erythroleukemic cells resulted in fusion-mediated insertion of viral antigens and of band 3 into the plasma membranes of the recipient cells. The presence of band 3 incorporated into Friend erythroleukemic cells was revealed structurally by electron microscopy after staining with ferritin-conjugated anti-human erythrocyte membrane antiserum. Incorporation of band 3, the putative erythrocyte antion channel, led to a marked and specific stimulation of anion exchange in Friend erythroleukemic cells. The viral-mediated insertion of an isolated membrane protein into viable Friend erythroleukemic cells provides a method for studying membrane protein turnover and for assaying reconstituted membrane components such as carriers or channels.


Binding of inhibitor alters kinetic and physical properties of extracellular cyclic AMP phosphodiesterase from Dictyostelium discoideum.

The extracellular adenosine 3',5'-cyclic monophosphate phosphodiesterase (3',5'-cyclic-nucleotide 5'-nucleotidohydrolase, EC 3.1.4.17) produced by Dictyostelium discoideum has two kinetic forms. The free enzyme has a Km of approximately 10 microM. The second form is the result of a complex formed with a heat-stable inhibitor and has a Km in the millimolar range. Treating the enzyme-inhibitor complex with dithiothreitol stimulated enzyme activity 20- to 100-fold and changed in Km from millimolar to micromolar. Dithiothreitol inactivated the inhibitor. Reconstituting purified enzyme with excess inhibitor returned the Km to the millimolar range. Under conditions known to inhibit the production of extracellular inhibitor or in mutants that lack it, extracellular phosphodiesterase activity was already high and could not be increased by dithiothreitol. The phosphodiesterase and inhibitor sedimented at 6 S and 3 S, respectively; the enzyme-inhibitor complex sedimented at 6.7 S.


[Triiodothyronine binding by the liver nuclei and mitochondria].

The binding of I125-triiodothyronine by male thyroidectomized rat liver nuclei and mitochondria in vivo and in vitro was studied. Labeled triiodothyronine was bound by the liver nuclei and mitochondria proteins. 90% radioactivity was bound by the nuclear nonhistone proteins. The binding of I125-triiodothyronine to the nuclei and mitochondria protein receptors was inhibited by unlabeled triiodothyronine and ICl. It is suggested that aromatic amino acids serve as the binding sites of the protein receptors, and that iodine atoms in the thyroid hormone molecules participated directly in the binding process.


Properties and localization of beta-endorphin receptor in rat brain.

Stereospecific high-affinity binding sites for beta h-[3H]endorphin could be demonstrated in the P2 pellet of rat brain homogenate. Scatchard analysis of the binding data revealed binding sites with Kd values of 0.81 and 6.8 nM and density of 120 and 240 fmol/mg of protein. Distribution of beta h-[3H]endorphin binding in various brain regions parallels that of opiate receptor:striatum greater than thalamus greater than amygdala greater than hypothalamus, septum greater than cortex greater than midbrain, brainstem. Similar to their effect on 3H-labeled agonist binding, Na+ and other monovalent cations, GTP, trypsin, chymotrypsin, phospholipase A2, and N-ethylmaleimide all inhibited the specific binding of beta h-[3H]endorphin. In contrast to their action on alkaloid and enkephalin binding, Ca2+, Mg2+, and Mn2+ also inhibited beta h-[3H]endorphin binding. These data suggest a difference between beta h-endorphin and alkaloid/enkephalin binding sites.


Purification of chicken intestinal receptor for 1,25-dihydroxyvitamin D.

The 3.3S chicken intestinal receptor for 1,25-dihydroxyvitamin D [1,25-(OH)2D] has been purified approximately 86,000-fold from the cytosolic fraction. The receptor was selectively precipitated with Polymin P from high-speed supernatants derived from 800 g of intestinal mucosa and then sequentially chromatographed on DNA-cellulose. Sephacryl, blue dextran-Sepharose, DNA-cellulose, and heparin-Sepharose. Polyacrylamide gel electrophoresis of 8-10 microgram of the purified receptor in sodium dodecyl sulfate indicated the presence of one major and three minor protein bands of molecular weights 50,000-65,000. Sucrose gradient analysis of the purified material in 0.3 M KCl suggested that a fraction of the receptor remained complexed to the 1,25-(OH)2D and that its sedimentation property of 3.3S remained unchanged. These results represent a major purification of the chick intestinal receptor for 1,25-(OH)2D, an extremely rare and labile protein whose isolation is estimated to require a 200,000-fold purification. Of primary importance is the observation tht affinity ligands such as DNA and blue dextran can effect major purification of this protein, lending credence to the hypothesis that the 1,25-(OH)2D receptor functions within the cell nucleus by altering the expression of specific genes.


Regulation of synthesis of a major outer membrane protein: cyclic AMP represses Escherichia coli protein III synthesis.

Cyclic AMP is the effector molecule for both positive and negative control of synthesis of several Escherichia coli proteins. Among the latter is the major outer membrane protein III. The control mechanism occurs at the level of transcription and involves the cyclic AMP receptor protein. The repressing system is saturated at lower concentrations of cyclic AMP than is the positive control system.


Complementation of the temperature-sensitive defect in H5ts125 adenovirus DNA replication in vitro.

Soluble extracts of adenovirus-infected HeLa cell nuclei support DNA replication on exogenous adenovirus DNA templates. Conditions of synthesis using both wild-type and temperature-sensitive extracts have been defined. Nuclear extracts prepared from cells permissively infected with the adenovirus mutant H5ts125 expressed the temperature-sensitive phenotype and could be inactivated at 37 degrees C in vitro. These extracts were completely complemented by the addition of wild-type adenovirus DNA binding protein but not by H5ts125 DNA binding protein. Enhancement by binding protein in the mutant extracts represents replication, as demonstrated by the production of full-sized products and orderly chain elongation originating, as in vivo, at both ends of the linear DNA. Replicative synthesis required the 5'-terminal protein bound covalently to template DNA and could be inhibited by denaturation of this 55,000-dalton protein. Various inhibitors of eukaryotic DNA polymerases, such as aphidicolin and 2',3'-dideoxythymidine triphosphate, inhibited replication of exogenous adenovirus templates in this system as they do in previously reported systems that only elongate endogenous replicating intermediates.


Spliced adenovirus-associated virus RNA.

We describe the structure of cytoplasmic RNA species transcribed from the DNA of adenovirus-associated virus, a defective parvovirus. The RNA was hybridized with minus strand template DNA and visualized in the electron microscope. Alternatively, the DNA.RNA duplex molecules were digested with nuclease S1 or Escherichia coli exonuclease VII and analyzed by agarose gel electrophoresis. A set of RNA species was observed with 5' terminal at map positions 5, 13, 19, or 39 and a 3' terminus and poly(A) tail at position 96 (one map unit is equivalent to 1% of genome length). Most of these RNAs are spliced and lack sequences approximately between positions 40 and 49. Some RNA preparations also contained unspliced molecules with 5' and 3' terminal at positions similar to those in the spliced RNA.


Solubilization of the low density lipoprotein receptor.

The low density lipoprotein (LDL) receptor was solubilized from membranes of bovine adrenal cortex and cultured human cells by incubation with the nonionic detergent octyl-beta-D-glucoside. Receptor activity released into the 100,000 x g supernatant was assayed by a solid-phase procedure: an aliquot of the soluble extract was removed, the detergent was diluted below its critical micellar concentration, causing the receptor to precipitate as a lipid-protein aggregate; the precipitate was collected by centrifugation and incubated with (125)I-labeled LDL ((125)I-LDL); and the receptor-bound (125)I-LDL was separated from free (125)I-LDL by filtration. The (125)I-LDL binding site that was precipitated from the soluble extract of bovine adrenocortical membranes appeared to be the same as the functional LDL receptor of cultured bovine adrenocortical cells and human fibroblasts. It exhibited high affinity and specificity (affinity for LDL more than 200-fold greater than for acetylated LDL, methylated LDL, or high density lipoprotein), dependence on calcium, and susceptibility to destruction by Pronase. The amount of (125)I-LDL binding activity in solubilized membranes from cultured cells was proportional to the number of receptors on the surface of the intact cells. Thus, the number of solubilized receptors was 1/20th of normal in mutant fibroblasts from a subject with homozygous familial hypercholesterolemia and was 1/4th of normal in human epithelioid carcinoma A-431 cells when they were grown in the presence of 25-hydroxycholesterol plus cholesterol. While in the soluble form in the presence of octyl-beta-D-glucoside, the LDL receptor can be carried through several steps of purification.


Synthesis of simian virus 40 t antigen in Escherichia coli.

Plasmids are constructed by using recombination in vitro according to Roberts, T.M., Kacich, R. & Ptashne, M. (1979) Proc. Natl. Acad. Sci. USA 76, 760-764 in which the t antigen gene of simian virus 40 is fused to a promoter of the Escherichia coli lac operon. In the fusions, transcription commences at the lac promoter, and, in some of the fusions, translation begins at the ATG initiator codon of the t gene. This translation is directed most efficiently by those plasmids in which the lac sequences abut the t gene such that a hybrid ribosome binding is encoded. In this case, the Shine-Dalgarno sequence is of lac origin but the ATG derives from the t gene. translation from this initiator codon is greatly decreased if the lac sequences are separated from the ATG by 17 base pairs and is abolished if the AT of this triplet is deleted. Cells bearing the productive fusions synthesize a 20,000-dalton protein with t antigenic determinants. This protein has an isoelectric point(s) indistinguishable from that of t antigen isolated from simian virus 40-transformed cells. Moreover, a partial sequence of the amino-terminal region of the bacterial product is that predicted for authentic t antigen. We conclude that these bacteria are for authentic t antigen. We conclude that these bacteria are producing a protein, the sequence of which is identical to that of authentic t antigen unfused to other polypeptides.


Differentiation as a requirement for simian virus 40 gene expression in F-9 embryonal carcinoma cells.

Infection of differentiated mouse embryo cells by simian virus 40 (SV40) leads to the production of the early mRNAs and the tumor (T) antigens that they encode. In contrast, undifferentiated F-9 murine teratocarcinoma cells do not support these early stages of the SV40 cycle. This block results from the inability to accumulate stable processed early SV40 mRNAs. It has recently been shown that vitamin A and its derivatives can induce in vitro differentiation of stem cells. Undifferentiated F-9 cells, upon treatment with a low concentration of retinoic acid, exhibited pronounced morphologic changes as well as the appearance of the H-2 surface antigens. After differentiation, the susceptibility of F9 cells to SV40 infection could be demonstrated by the appearance of large T and small T antigens, as shown by immunofluorescence and immunoprecipitation. Furthermore, SI nuclease mapping of early SV40 transcripts confirmed the presence of the two spliced early mRNAs. These results indicate that the undifferentiated F-9 stem cells contain the genetic information needed for generating stable processed early SV40 mRNAs but are blocked in the production of functional species.