Peripheral neuropathy and benign IgG paraproteinaemia. Three patients with peripheral neuropathy and an associated benign IgG paraproteinaemia are described. No direct immunological evidence for an aetiological role of the paraprotein was found, and the implications of this are discussed.


Electronystagmographic criteria in neuro-otological diagnosis. 1. Peripheral lesions. Direct current electronystagmographic recordings of spontaneous nystagmus, in the light, in darkness, and with eye closure were carried out on 34 patients with peripheral lesions of the vestibular system with a view to determining their specificity. In 60% of patients the responses were characterised by an enhancement of the nystagmus both with eye closure and in darkness. The remainder exhibited no nystagmus under any recording condition.


[Peripheral neuropathy with cryoglobulinemia (author's transl)]. The author describe 2 personal observations of peripheral neuropathy with cryoglobulinemia and the 28 cases previously recorded are reviewed. The characteristics of the usually sensorimotor neuropathy are not specific. Nevertheless, the association with purpura, Raynaud's syndrome and leg ulcers and the inconstant aggravation of the symptoms with cold allow the diagnosis to be suspected on clinical grounds. Cryoglobulin can be recognised by immunoelectrophoresis and classified as type I monoclonal, types II and III mixed and polyclonal. Associated disease should be looked for; lymphocytic proliferation or auto-immune disease. If none is found a diagnosis of essential cryoglobulinemia can be made but with caution as cryoglobulinemia can precede by several years the appearance of associated disease.


Moebius syndrome, peripheral neuropathy and hypogonadotrophic hypogonadism. The association of congenital ophthalmoplegia and facial paresis (Moebius syndrome) with a variety of other developmental somatic defects has been widely recognised. Its co-existence with hypogonadism of hypothalamic/pituitary origin and subclinical peripheral neuropathy has been reported and in this paper we describe the second case of the Moebius syndrome in association with hypogonadotrophic hypogonadism and a progressive peripheral neuropathy of mixed axonal and demyelinating type.


How do axons control myelin formation? The model of 6-aminonicotinamide neuropathy. Injection of 6-aminonicotinamide into young rats produces a peculiar neuropathy characterized by selective swelling and disruption of the layer of Schwann cell cytoplasm lining the inner surface of the myelin sheath. This layer increases greatly in volume, compressing the axon and distending the myelin sheath. Morphometry of such swollen fibers discloses that the amount of myelin in the distended sheaths is considerably greater than would correspond to the size of the axons, even if axonal compression is accounted for. The data favor the concept that sheath growth is stimulated by non-specific distension of the myelin sheath from inside.


Fatty acid accumulation and abnormal lipid deposition in peripheral and border zones of experimental myocardial infarcts. Twenty-eight dogs with acute anterior myocardial infarcts due to proximal occlusion of the left anterior descending coronary artery (LAD) were studied at various periods following the occlusion to determine: (a) the time course and location of abnormal lipid accumulation after infarction, (b) the degree of muscle-cell injury associated with increased lipid deposition, and (c) whether uptake of fatty acid from the circulating fat pool contributes to lipid accumulation in certain myocardial regions. The findings show that myocardial lipid accumulation begins as early as 6 hr after proximal LAD occulsion. The increased lipid deposition occurs as nonmembrane-bound lipid droplets in muscle cells with and without ultrastructural evidence of irreversible injury. Analysis of tissue uptake of intravenoulsy injected [14C] oleic acid conjugated with albumin revealed relatively selective concentration of label in the peripheral and border regions of the infarct, but occasionally even the central subendocardial portion of the infarct concentrated the fatty acid. Thin-layer chromotography showed that most of the label was associated with the triglyceride fraction when the radiolabeled fatty acid was injected 6 or 24 hr after LAD occlusion. These myocardial cellular and topographical alterations will have to be considered when labeled fatty acids are used for imaging acute myocardial infarcts and/or if attempts are made to identify myocardial fat-laden cells scintigraphically.


Tc-99m pyrophosphate muscle labeling in McArdle syndrome. This paper reports the findings on two patients with McArdle syndrome (myophosphorylase deficiency) in whom conventional bone scans with Tc-99m pyrophosphate revealed intense muscle labeling following exercise tests. The temporal pattern observed was similar to that seen with other types of muscle damage. The prolonged cramps often occurring with this entity appears to produce muscle damage that is readily demonstrable using conventional bone-scanning techniques.


Scintigraphic abnormalities in glycogen storage disease. Fifteen patients with glycogen-storage disease type 1 (von Gierke's disease) were evaluated by serial scintigraphy, with a clearly recognizable pattern of an enlarged liver with diminished radionuclide accumulation, splenomegaly with considerably increased uptake and renomegaly. In seven of these patients with GSD-1 scintigraphy demonstrated focal defects of varying size. Small or stable defects suggest benign hepatic adenomata, whereas malignant change occurred in growing large lesions. The potential malignant end-point of hepatic-cell carcinoma in GSD-1 warrants careful serial liver scintigraphy with scintiangiography on a routine basis.


Enteroviral disease in early infancy.

Enteroviral disease in early infancy. To discover the nature of enteroviral infections in early infancy (infants less than two months of age) in the United States, data were examined of 338 such infants with nonpolio enterovirus isolates reported to the Center for Disease Control in 1972-1975. Interpretation of such data is limited by reporting bias and difficulty in documenting disease causality. Enteroviral disease was severe in 74% of the infants. The most common clinical manifestations were meningitis, severe systemic disease, nonspecific febrile illness, and gastrointestinal symptoms. Fifty-one percent of all enteroviral disease was associated with echoviruses; 45% with Coxsackie B viruses; and only 4% with Coxsackie A viruses. Disease was more prevalent in the summer months, was recognized throughout the country, and occurred more frequently in males.


Archenteronoma (yolk sac tumors).

Archenteronoma (yolk sac tumors). Ten cases of archenteronoma are reviewed with a follow-up between 2 mo and 7 yr. Their presentation was similar to that of rhabdomyosarcoma except for testicular tumors that may present as a hydrocele. A good prognosis was found with tumors amenable to primary resection and chemotherapy. Other tumors are best treated initially with chemotherapy before relatively conservative surgery, radiotherapy being used either pre- or postoperatively.


Behavioral and electrophysiological effects of peptides related to lipotropin (beta-LPH). Both C-terminal fragments of lipotropin (beta-LPH) (endorphins) and N-terminal fragments (e.g., ACTH 4-10) delayed extinction of pole-jumping avoidance behavior in rats. After subcutaneous injection Met5-enkephalin appeared to be as active as ACTH 4-10 whereas beta-LPH 61-69, alpha- and beta-endorphin were more potent in delaying extinction  of pole-jumping avoidance behavior (approximate ED50 of alpha-endorphin 4 x 10(-11) M rat.) However, the potency of beta-LPH 61-69 and alpha-endorphin appeared to be approximately the same whereas that of beta-endorphin was less than that of ACTH 4-10 after intraventricular administration (approximate ED50 of alpha-endorphin 0.2 x 10(-11) M rat). alpha-Endorphin and ACTH 4-10, administered subcutaneously in a dose which markedly delayed extinction of pole-jumping avoidance behavior, had only slight effects on open field behavior and on responsiveness to electric footshock. A 5 times higher dose of both peptides facilitated passive avoidance behavior. Morphine in two doses significantly delayed extinction of pole-jumping avoidance behavior but the effect was not dose dependent. The specific opiate antagonist naltrexone, however, markedly facilitated extinction of the avoidance response. ACTH 4-10, alpha- and beta-endorphin and a behaviorally potent ACTH 4-9 analog (Org 2766) restored pole-jumping avoidance behavior of rats pretreated with naltrexone. Treatment with a similar dose of naltrexone blocked beta-endorphin-induced analgesia. These results suggest that the influence of peptides related to C-terminal and N-terminal fragments of lipotropin on extinction of avoidance behavior may be dissociated from those exerted on opiate receptor sites. Subcutaneously injected beta-LPH 61-69 or intraventricularly administered beta-endorphin induced a shift from lower to higher frequencies of hippocampal theta rhythm during paradoxical sleep in the same way as that found after ACTH 4-10. This effect is interpreted as indicating an increased arousal state in certain midbrain limbic structures. This may, as has been postulated for ACTH 4-10, alter the motivational value of environmental stimuli (e.g., aversive stimulation).


Purine nucleoside and nucleotide interactions on normal and subsensitive alpha adrenoreceptor responsiveness in guinea-pig vas deferens. Adenosine and adenosine 5'-monophosphate (AMP) augment contractile responses to norepinephrine (NE) in isolated guinea-pig vas deferens. Dipyridamole slightly enhances, while theophylline antagonizes, adenosine effects on responses to NE. Adenosine triphosphate (ATP) and the nonhydrolyzable analog, adenosine 5'-(beta,gamma-imido)triphosphate (AppNp) depress responses to NE. Adenine and adenosine diphosphate (ADP) are ineffective in influencing alpha adrenoreceptor responsiveness. Repetitive stimulation of isolated vas deferens with maximal concentrations of NE markedly reduce the contractile response to test concentrations of 6 micron NE. Spontaneous resensitization of responses to NE to control levels occurs within 25 to 35 minutes after the end of desensitization treatment. Adenine nucleosides and nucleotides promote a more rapid rate of alpha adrenoreceptor resensitization, with a potency order: AMP greater than adenosine greater than ADP. Adenine and ATP did not influence the rate of alpha adrenoreceptor resensitization. The adenine nucleotides ADP, ATP and the analog AppNp elicit concentration-dependent contractions of guinea-pig vas deferens. Theophylline antagonizes this contractile activity to adenine nucleotides. AMP, adenosine and adenine are devoid of agonistic activity. In the presence of NE, however, AMP and adenosine produce contractile responses of isolated vas deferens strips, and the agonistic activity of ADP, ATP and AppNp is profoundly enhanced. Agonistic actions of purines in the presence of NE are antagonized by phentolamine much more effectively than by theophylline. The results suggest the existence of a purinergic receptor mediating excitatory responses of guinea-pig vas deferens. Furthermore, there appears to be mutual interaction between purinergic and alpha adrenoreceptor mechanisms. That adenyl derivatives are capable of augmenting subsensitive alpha adrenoreceptor responsiveness suggests that adenine nucleosides or nucleotides, released during sympathetic transmission, may be required for maintenance of normal alpha adrenoreceptor sensitivity.


Epinephrine-induced automaticity of canine cardiac Purkinje fibers and its relationship to the adenylate cyclase-adenosine 3',5'-monophosphate system. We studied the relationship between epinephrine-induced increases in automaticity and in the adenylate cyclase-adenosine 3',5'-monophosphate (cAMP) system in canine cardiac Purkinje fibers. In intact Purkinje fiber bundles superfused with Tyrode's solution, epinephrine induced a concentration-dependent increase in automaticity and in cAMP content. Both of these effects were reduced by propranolol. Decreasing the temperature of the superfusate from 37 degrees to 25 degrees C blocked the epinephrine-induced increase in automaticity, but not the increase in cAMP content. Addition of the metabolic blocking agent, iodoacetate, to the superfusate did not block the effect on Purkinje fiber automaticity, but the cAMP content did not increase. In Purkinje fiber bundle whole homogenates adenylate cyclase was stimulated more by isoproterenol than by epinephrine. Low concentrations of phenylephrine decreased adenylate cyclase activity; higher concentrations induced an increase toward control values. Iodoacetate did not significantly alter cyclase activity and did not affect its response to epinephrine. These students have shown that although intact Purkinje fiber bundles respond to epinephrine with increases in automaticity and cAMP content, these two events can be dissociated under appropriate conditions.


Position sensitivity of de-efferented muscle spindles in experimental acrylamide neuropathy. An experimental neuropathy was induced in cats by injections of acrylamide (7.5, 15 or 30 mg/kg/day) for 2 to 10 days. The responses of primary and secondary endings of soleus muscle spindles to stretch were evaluated and correlated with the appearance of ataxia and incoordinated motor movements. Animals that received 15 or 30 mg/kg/day became ataxic and demonstrated poor motor coordination on the 7th or 4th day, respectively. At these times, both primary and secondary endings of muscle spindles had elevated thresholds and diminished discharge frequencies. Continued acrylamide administration resulted in exacerbation of the clinical symptoms and further attenuation of spindle responses. The discontinuation of acrylamide was followed by slow recovery. Only those cats which received a total dose of 75 mg/kg or less remained asymptomatic and had normal spindle function. The coincidence of onset of motor coordination deficits and spindle dysfunction, coupled with a lack of demonstrable motor defect at the same time, suggests that the initial clinical features of acrylamide neuropathy may be partly the consequence of impaired spindle function.


Involvement of nicotinic and muscarinic receptors in synaptic transmission in cat superior cervical ganglions reinnervated by vagal primary afferent axons. Artificial synapses were established in the superior cervical ganglion reinnervated by vagal afferent fibers by heterologous cross-anastomosis between the cranial end of nodose ganglion and the caudal end of superior cervical ganglion in cats. Formation of functional synapses was evidenced by unilateral mydriasis and contraction of the nictitating membrane in response to inflation of the stomach with a balloon or to electrical stimulation of the afferent vagus. Electron microscopic findings indicated that the vagal afferent fibers terminated in the superior cervical ganglion after cross-anastomosis. In the superior cervical ganglion reinnervated by the afferent vagus, activities of choline acetyltransferase and cholinesterase were higher than those in the preganglionically denervated ganglion, but lower than those in the sympathetic preganglionically reinnervated ganglion. Contractions of the nictitating membrane and postganglionic action potentials evoked by electrical stimulation of the vagal artificial preganglionic trunk in the cross-anastomosed ganglion were blocked by treatment with tetraethylammonium and also with atropine. Atropine did not affect these responses in the normal and the preganglionically reinnervated ganglion, except at an early stage after operation. Comparisons of pharmacological properties in normal, anastomosed, preganglionically denervated and reinnervated ganglia indicated that activation of muscarinic receptors in the anastomosed ganglia is probably not secondary to an incomplete nerve supply, but may be dependent on the nature of the nonmyelinated vagal afferent fibers. The possibility that the transmitter involved may be acetylcholine is discussed.


Effects of theophylline and N6,O2-dibutyryl adenosine 3':5'-monophosphate on sympathetic ganglionic transmission in rats. The effects of theophylline and N6,O2-dibutyryl adenosine 3':5'-monophosphate (DBcAMP) on the amplitude of the postganglionic action potential during and after a 10 Hz repetitive volley, and 50 to 1000 msec after a conditioning stimulus were investigated. The effects of both drugs on some electrophysiological properties of single cells of the isolated superior cervical ganglia of rats were also studied. At low concentrations of theophylline a reversible potentiation of the compound action potential occurred during and after repetitive stimulation at 10 Hz and also after the single conditioning stimulus. This effect was antagonized by atropine. Large concentrations of theophylline exerted a depressive effect only. Low concentrations of DBcAMP caused a reversible initial depression followed by a durable facilitation of transmission during repetitive stimulation. These concentrations potentiated the action potential amplitude after repetitive stimulation, but depressed it after a single conditioning stimulus. Atropine augmented the latter two effects. DBcAMP at large concentrations depressed transmission, but transmission was facilitated after drug washout. Theophylline and guanosine 3':5'-monophosphate, at ineffective concentrations when used singly, potentiated each other and elicited facilitation which was abolished by atropine. Theophylline and DBcAMP at these concentrations depolarized ganglion cells with a time course shorter than that of the aforementioned effects. Both drugs reduced the frequency and amplitude of the spontaneous miniature excitatory postsynaptic potentials. Theophylline did not increase the evoked transmitter release appreciably. On the basis of these findings and the evidence from literature, it is suggested that the reversible facilitatory effect of theophylline may be at least in part due to inhibition of phosphodiesterase of the ganglion cells leading to an enhanced muscarinic transmission. The prolonged facilitatory effect of DBcAMP may result from a durable change in the postsynaptic membrane structure leading to enhanced muscarinic transmission. An enhancement in the muscarinic transmission by both drugs increases the membrane excitability causing recruitment of subthreshold depolarized cells to discharge resulting in facilitation.


Control of acetylcholine sensitivity and synapse formation by muscle activity. 1. The formation of ectopic junctions between the 'foreign' superficial fibular nerve and the soleus muscle of adult rats, and its relation to changes in extrajunctional sensitivity to acetylcholine (ACh), has been studied by denervating the muscle 3-6 weeks after implanting the foreign nerve. 2. The earliest signs of nerve-muscle transmission were seen 2.5-3 days after denervation, in those fibres where the extrajunctional ACh sensitivity first reached its full post-denervation level. The number of innervated fibres continued to increase throughout the first week after denervation until 70-100% of fibres underlying the foreign nerve growth were innervated. 3. Direct stimulation of muscles with chronically implanted electrodes from the time of denervation prevents the formation of functional neuromuscular junctions (n.m.j.s). If stimulation begins 2 or 4 days after denervation, some functional n.m.j.s are formed which can be detected 7-9 days after denervation, though not as many as in the absence of stimulation. 4. Direct stimulation of muscles from the time of denervation prevents the development of detectable extrajunctional ACh sensitivity. If stimulation begins 2 days after denervation nearly maximal sensitivity develops during the third day and then rapidly declines to undetectable levels by the beginning of the eight day after denervation.


Cyclic AMP and cyclic GMP in rabbit blastocysts. Concentrations of both nucleotides were significantly higher in Day-6 than in Day-5 blastocysts but the ratio of cAMP to cGMP changed from 0.5 to 1.5.


The relationship of puberty to other maturity indicators and body composition in man. The purpose of this paper is to examine the relationship between sexual development and other maturational processes in children. The word "puberty" is used as a general term to include the development of the secondary sex characters and the attainment of reproductive competence.


Effect of polymyxin B on the structure and the stability of lipid layers. Polymyxin B (PX) does not penetrate phospholipid monolayers and bilayers at low field strength across the lipid layers. The degree of penetration of PX is evaluated from its effect on the capacitance of the monolayers and on the conductance of the bilayers. PX added to one side of a bilayer causes its destabilization, it also enhances destabilization of lipid monolayers at positive electric fields across the surface layer in the direction of the adsorbed PX. PX lowers very little the fluorescence polarization of 1,6-diphenyl 1,3,5 hexatriene embedded in phospholipid vesicles. It is suggested that the penetration mechanism of PX into gram-negative bacteria is based on transient local breakdown of the plasma membrane.


Mode of action of theophylline on sodium efflux in barnacle muscle fibers. The response of the Na efflux in unpoisoned barnacle fibers to 10 mM theophylline is biphasic; i.e., inhibition is followed by stimulation. The stimulatory response is unaffected by ouabain. Fibers pretreated with ouabain show no transitory inhibition when 10 mM theophylline is applied, but show prompt stimulation the magnitude of which is comparable to that observed with unpoisoned fibers. The same holds true for lower concentrations of theophylline. Prior injection of 500 mM EGTA completely abolishes the biphasic action of 10 mM theophylline. External application of 10 mM theophylline following removal of external Ca2+ fails to bring about a biphasic effect. Ca2+ restoration, however, results in a moderate rise in the Na efflux. External application of 10 mM theophylline stimulates the Na efflux into Ca2+-free artificial seawater (ASW) when the test fibers are pretreated with ouabain. Injection of the protein inhibitor of Walsh leads to reduced stimulation by 10 mM theophylline of the Na efflux in unpoisoned fibers. Injection of the protein inhibitor of Corbin into unpoisoned fibers leads to reduced stimulation by 10 mM theophylline. Injection of cAMP into ouabain-poisoned fibers, following internal application of Corbin's inhibitor and external application of 10 mM theophylline, fails to cause a marked rise in the ouabain-insensitive Na efflux. Injection of Corbin's inhibitor into ouabain-poisoned fibers, following the onset of peak stimulation by 10 mM theophylline, fails to reduce the Na efflux. Fibers injected with 1 mM and 100 mM EGTA and exposed to 10 mM theophylline show a marked reduction in the response of the ouabain-insensitive Na efflux to injected cAMP when the concentration of theophylline is 10 mM. A poor response to injected cAMP is also seen in fibers bathed in Ca-free ASW containing 10 mM theophylline. Theophylline (10 mM) fails to cause an enhanced stimulation of the ouabain-insensitive Na efflux into Ca-free 3 mM-HEPES ASW or 10 mM-Ca2+ -3mM-HEPES ASW following the addition of protons to the bathing medium. An enhanced response is similarly not observed with injected cAMP following the addition of theophylline to the bathing medium. Injection of 8-fluorotheophylline, 3-isobutyl-1-methylxanthine and doxantrazole leads to a marked reduction in the response of the ouabain-insensitive Na efflux to injected cAMP. Contraction always takes place upon injecting these substances. These results are in keeping with the theory that theophylline acts chiefly by reducing myoplasmic pCa(pCa=-log10[Ca2+]), and that a reduced pCa leads to stimulation of the ouabain-insensitive Na efflux as the result of activation of the cGMP-dependent protein kinase system by newly formed cGMP.


Effect of basic protein from human central nervous system myelin on lipid bilayer structure. The effect of myelin basic protein from normal human central nervous system on lipid organization has been investigated by studying model membranes containing the protein by differential scanning calorimetry or electron spin resonance spectroscopy. Basic protein was found to decrease the phase transition temperature of dipalmitoyl phosphatidylglycerol, phosphatidic acid, and phosphatidylserine. The protein had a greater effect on the freezing temperature, measured from the cooling scan, than on the melting temperature, measured from the heating scan. These results are consistent with partial penetration of parts of the protein into the hydrocarbon region of the bilayer in the liquid crystalline state and partial freezing out when the lipid has been cooled below its phase transition temperature. The effect of the protein on fatty acid chain packing was investigated by using a series of fatty acid spin labels with the nitroxide group located at different positions along the chain. If the protein has not yet penetrated, it increases the order throughout the bilayer in the gel phase, probably by decreasing the repulsion between the lipid polar head groups. Above the phase transition temperature, when parts of it are able to pentrate, it decreases the motion of the lipid fatty acid chains greatly near the polar head group region, but has little or no effect near the interior of the bilayer. Upon cooling again the protein still decreases the motion near the polar head group region but increases it greatly in the interior. Thus, the protein penetrates partway into the bilayer, distorts the packing of the lipid fatty acid chains, and prevents recrystallization, thus decreasing the phase transition temperature. The magnitude of the effect varied with the lipid and was greatest for phosphatidic acid and phosphatidylglycerol. It could be reversed upon cooling for phosphatidylglycerol but not phosphatidic acid. The protein was only observed to decrease the phase transition temperature of phosphatidylserine upon cooling. It had only a small effect on phosphatidylethanolamine and no effect on phosphatidylcholine. Thus, the protein may penetrate to a different extent into different lipids even if it binds to the polar head group region by electrostatic interactions.


Cytotoxic activity of methylene bis-nucleotides. New facts relating to the mechanism of cytotoxic and antitumor activity of formaldehyde-treated RNA and its hydrolysates have been revealed. Synthetic methylene bis-nucleotides and, in particular, methylene bis-nucleosides have a cytotoxic effect on cultured human adenocarcinoma cells (line HCCL). The cytotoxic effect of methylene bis-derivatives of the guanine series was most pronounced. In addition, methylene bis-nucleotides were depbosphorylated in the cell culture medium. Chromosome aberrations were induced by alkaline hydrolysate of dialyzed formaldehyde-treated RNA in which 39% of purine nucleotdies were modified to methylene bis-derivatives.


Effect of the Fv-1 locus in vivo: host range pseudotypes of murine sarcoma virus. Fv-1-specific host-range pseudotypes of murine sarcoma virus (MuSV) were developed by rescue from nonproducer cells with N- or B-tropic leukemia viruses. The MuSV(B) and MuSV(N) pseudotype viruses were tested in vitro and were restricted specifically by the Fv-1 gene locus. When the pseudo-type viruses were tested in vivo in mice of specific Fv-1 geno-types, tumor induction was completely restricted in nonpermissive animals, including athymic nude mice, whereas tumors grew progressively in permissive animals. All tumors regressed in adult heterozygous mice, but not in adult athymic nude mice.


Simian virus 40 (SV40) production from SV40-transformed human amnion cells of established lines. Sixteen established cell lines of simian virus 40 (SV40)-transformed human amnion cells were examined for SV40 production. Many of these lines produced SV40 for extensive periods. Virus production had not ceased for 2 lines after 18 months, for 3 lines after 12 months, and for 3 lines at 3 months after recovery from "crisis". Three lines became virus-free in the first month, 1 line in the second month, 1 in the third month, and 1 in the fourth month, and 2 lines stopped virus production between 6 and 11 months after recovery. The virus titers were relatively low. Inclusion body-containing cells were infrequent. In contrast, in most cultures of SV40-transformed human fibroblasts rescued from crisis, no infectious virus was demonstrated, although exceptions have been reported. Virus was produced after heterokaryon formation of cells of the virus-free amnion lines with CV-1 cells in the presence of inactivated Sendai virus, as observed for SV40-transformed human fibroblasts. During the crisis period, some of the SV40-transformed amnion cells produced substantial amounts of virus. Titers decreased during the later periods of crisis. The most pronounced decrease in titers was in cultures from which established lines were recovered.


Induction of intracisternal type A particles by 5-bromo-2'-deoxyuridine in rat hepatoma cells. Electron microscopic examination of hepatoma tissue culture (HTC) cells revealed low numbers of intracisternal type A particles (IAP) and type C viruses. Exposure of HTC cells to either 10(-4) or 10(-5) M5-bromo-2'-deoxyuridine (BUdR) caused a more than 50-fold increase in the number of IAP observed with the electron microscope. The number of IAP increased after only 2 days of growth in 10(-5) M BUdR, whereas 3 days of growth in 10(-4) m BUdR were necessary to observe an increase. A 2-day pulse of 10(-4) M BUdR was also sufficient to cause the increase in type A particles, provided the cells were continued in culture for another 2 days. Unlike many other cell lines, HTC cells treated with BUdR did not show an increase in type C viruses. This conclusion was based on the observations that BUdR treatment caused no detectable increase in extracellular particulate viral reverse transcriptase or in viral RNA complementary to a DNA probe made to a rat endogenous type C virus (RaLV).


Variation of properties of chrysotile asbestos subjected to milling. Mechanical milling is commonly used to produce short chrysotile asbestos for experimental purposes. Such manipulation also decreases fiber crystallinity, alters Si-O and Mg-O interlayer bonding, induces coordination changes in the brucite layer, diminishes the ability of fiber to reduce specific free radicals and physisorb organic molecules, and decreases hemolytic potency and antagonist sorption capabilities. The degree of alteration is related to the time of milling. Results of biological experimentation with these materials must be interpreted with caution. Interaction mechanisms in the biological setting are suggested for chrysotile fiber.


Latex agglutination test for amoebiasis in Pakistani patients with chronic 'obscure' liver enlargements. The latex agglutination test for amoebiasis was done in 50 Pakistani patients in whom a clinical diagnosis of chronic non-supparative amoebic hepatitis was made on the basis of liver enlargement associated with chronic recurrent abdominal disturbance. The serological test was negative in 32 (64%) and positive in 18 (36%). Examination of stool specimens showed a higher frequency of E. histolytica in serology positive patients. The liver biopsy findings were variable and included normal histology in 40 percent and non-specific changes in another 34 per cent of the patients. There was evidence of early abscess formation in one patient with a strongly positive serological reaction. Chronic liver enlargements in this region form a heterogenous group and the existence of chronic non-suppurative amoebic hepatitis as a clinico-pathological entity remains doubtful.


Toxoplasmosis, distemper, and herpesvirus infection in a skunk (Mephitis mephitis). A striped skunk (Mephitis mephitis) showing abnormal behavior had histopathologic lesions of toxoplasmosis and canine distemper in addition to intranuclear, eosinophilic inclusions in the reticuloendothelial cells of the spleen, liver and lung. The inclusions, on electron microscopic examination, were compatible with herpesvirus infection.


Oral contraceptives and nonfatal myocardial infarction. We obtained information on 107 women younger than 46 years who were discharged from a hospital with a diagnosis of acute myocardial infarction. In the series 26 women were otherwise apparently healthy and potentially childbearing. Among these 26 women, 20 (77%) were taking oral contraceptives just prior to admission, and one was taking conjugated estrogens. Among 59 control women, 14 (24%) were taking oral contraceptives and one was taking conjugated estrogens. The relative risk estimate, comparing oral contraceptive users with nonusers, is 14 with 90% confidence limits of 5.5 and 37. All but two of the 26 women were cigarette smokers. While this illness is rare in most healthy young women, the risk in women older than about 37 years who both smoke and take oral contraceptive appears to be high.


Noncontraceptive estrogens and nonfatal myocardial infarction. We obtained information on 107 women younger than 46 years discharged from a hospital with a diagnosis of acute myocardial infarction. In the series there were 17 women aged 39 to 45 years who were otherwise apparently healthy and had had a natural menopause, hysterectomy, or tubal ligation or whose spouse had had a vasectomy. Among them, nine (53%) were taking noncontraceptive estrogens just prior to admission. Among 34 control women, four (12%) were taking estrogens. The relative risk estimate, comparing estrogen users with nonusers, is 7.5, with 90% confidence limits of 2.4 and 24. All but one of the 17 ml subjects were cigarette smokers. While this illness is rare in most healthy young women, the risk in women older than about 38 years who both smoke and take estrogens appears to be substantial.


Renal tranplantation in type 1 glycogenosis. Failure to improve glucose metabolism. In glycogenosis type 1 (GT1), glucose synthesis is deficient due to absence of glucose-6-phosphatase. Development of renal failure in such a patient provided the opportunity to test whether or not this metabolic defect could be reversed by a renal allograft, which contains the missing enzyme and has potential for glucose synthesis. Despite normalization of renal function and both glucocorticoid therapy and the infusion of amino-acid precursors of glucose, fasting hypoglycemia persisted unabated. We conclude that a funtioning renal allograft is incapable of meeting the metabolic demands of a patient with glucose-6-phosphatase deficiency.


Possible involvement of cyclic AMP in inflammation induced by a surfactant. Alkyldimethylbenzylammonium chloride (alkyl-DBAC), a cationic surfactant, produces acute exudative inflammation accompanied by an enhancement of energy metabolism. The mechanism of metabolic changes, consequently the cyclic AMP level and the effects of certain drugs were determined in the gastrocnemius muscles of rats in which acute exudative inflammation had been induced by intramuscular injections of alkyl-DBAC. A transient increase in the cyclic AMP level was noted at 15 to 30 minutes after the injection of alkyl-DBAC, and this elevation was antagonized by chlorpromazine, diphenhydramine, promethazine, aspirin and indomethacin. The time course of increasing tendency in the cyclic AMP level after the injection of histamine closely paralleled that of alkyl-DBAC. These results suggest that cyclic AMP may be involved in the metabolic changes with inflammation induced by alkyl-DBAC.


Studies on beta-pyridylcarbinol as a precursor of nicotinamide adenine dinucleotide. The major part of orally administered beta-pyridylcarbinol was converted to nicotinic acid in the gastrointestinal tract, whereas that administered intraperitoneally was partially converted to nicotinic acid in the liver. The conversion of this compound to nicotinic acid in both the gastrointestinal tract and liver involved enzymatic processes. The converted nicotinic acid from beta-pyridylcarbinol may play a dominant role in the biological actions of beta-pyridylcarbinol and possibly involves in the synthesis of NAD via Preiss-Handler's pathway.


[Effect of neural regulation on the fluctuations in the intervals between cardiac contractions and the force of the contractions]. Statistical analysis of the cardiac rhythm according to the R--R intervals and the force of contractions according to their amplitude was undertaken. The variation coefficient (N%) reflecting the degree of irregularity of intervals between the contractions and the force of contractions during maximum cardiac hyperfunction induced by clamping the aorta for 30 sec was determined. Vagotomy led to a decrease in the irregularity for cardiac rhythm and force of cardiac contractions.


[Histology, fine structure and differentiation of experimental Wilms tumors (author's transl)]. Histologic studies indicate that the cycasin-induced Wilms' tumor in the rat is equal to human nephroblastoma in appearance. Therefore, it may represent an interesting system for experimental oncology. In electron microscopy, the spindle cells of the sarcoma region mostly represent tubular cells. Besides, the author's results showed that preexisting mesenchyma cells and blood vessels also form part of the tumor. Electron microscopy studies have shown that cycasin feeding will lead to early cellular changes indicating that cycasin exercises a nucleotoxic effect. Present results do not favor the dysontogenic concept of the formation of Wilms' tumor, since this tumor can be induced diaplacentally.


[Inguinal Wilms' tumor (author's transl)]. Case report on a small inguinal Wilms' tumor in a 3-year-old boy. Their is strong evidence, that this heterotopic Wilms' tumor developed in dysplastic renal tissue.


Control of plasma aldosterone in diabetic patients with hyporeninemic hypoaldosteronism. In three patients with diabetes and hyporeninemic hypoaldosteronism changes in renin activity, plasma aldosterone and cortisol were examined under various conditions: orthostasis and intravenous furosemide, infusion of synthetic beta1-24 ACTH on two consecutive days and diurnal variations in basal hormone fluctuations. Each patient showed unmeasurably low renin activity unresponsive to orthostasis and intravenous furosemide while plasma aldosterone was below normal range. Under ACTH-infusion only marked increases in aldosterone were observed in one patient whereas cortisol responded normally in all diabetics tested. Analysis of diurnal night day fluctuations (20.00-8.00) in plasma aldosterone and cortisol revealed a close and statistically significant relationship between both hormones in each of the three patients (p less then 0.05-less than 0.001). Variations in plasma aldosterone thus were mediated through changes in endogenous pituitary ACTH. Compared with normal controls however, diurnal aldosterone curves were set at a lower level. Our results demonstrate that a reduced sensitivity of the adrenal gland to ACTH is not responsible for the observed subnormal plasma aldosterone levels in these patients. Therefore, the lack of circulating angiotensin II seems to be the causative reason of hypoaldosteronism. The exact mechanism of undetectable renin activity in these patients remains unknown.


In vitro demonstration of Mallory body formation in liver cells from rats fed diethylnitrosamine. Epithelioid liver cells were established in culture from rats sacrificed 10 to 18 weeks after the administration of the hepatocarcinogen diethylnitrosamine in their drinking water. After approximately 3 months in vitro, more than half the cells propagated from rats that developed hepatocellular carcinomas had bean-shaped, acentrically displaced nuclei with large juxtanuclear homogeneous appearing areas resembling the hyalin or Mallory bodies in the livers of chronic alcoholics. These abnormalities were not seen in the livers of origin, but were retained in the carcinomas that formed after the cultured cells containing such juxtanuclear hyalin inclusions were inoculated into young rats or nude (i.e., thymusless) mice; these features persisted upon reestablishment and continuous passage of the tumor cells in culture. The cells were further characterized by their karyotypes and their growth properties in liquid media and soft agar. By transmission electron microscopy the hyaline bodies in the culture tumor cells were shown to consist of a disorganized meshwork of filaments. Examination by incubating cells with cytochalasin B and by using antiactin and anti-DNase antisera as indirect immunofluorescence probes also revealed a disturbance in the cytoskeleton.


Effects of chronic and acute ethanol administration on sleep in laboratory rats. Long- and short-term administration of nonintoxicating doses of ethanol affected sleep parameters in rats.


Evaluation of a national training program for consultants in alcoholism treatment. A method for evaluating the training of persons hired to develop alcoholism treatment programs in government and private industry is described.


Cyclic nucleotide levels in resting and mitogen-stimulated spleen cell suspensions from young and old mice. The levels of cyclic adenosine 3', 5'-monophosphate (cAMP) in suspensions of unstimulated spleen cells from tumor-free 30-month old (C57BL/10Sn X C3H/HeDiSn)F1 hybrid mice averaged only 14% of that of 6-month old mice. By contrast, the level of cyclic guanosine 3', 5'-monophosphate (cGMP) in spleen cell suspensions from old mice was about 270% that of young mice. The cAMP/cGMP ratio for the unstimulated (resting) state showed a decline by 30 months to about 5% of its 6-month value. Cyclic nucleotide levels were also measured in cell suspensions from old and young mice at intervals over a two hour period following in vitro stimulation with the plant mitogens phytohemagglutinin, concanavalin-A and pokeweed mitogen. Quantitative and in some instances qualitative differences in responses were noted. These results might conceivably reflect either age-related changes in the splenic lymphoid cell subpopulations or intrinsic cellular alterations or both. It is unlikely that changes of this degree could be wholly explained by population shifts. An imbalance in cyclic nucleotide levels in both resting and stimulated lymphoid cells in older animals might contribute to the immune dysfunction known to occur with normal aging.


[Hepatic fibrosis--mechanism, dynamics and clinical consequences (author's transl)]. Hepatic fibrosis may result from collapse after hepatocellular necrosis or from new formation of connective tissue. Fibroplasia, particularly within the lobular parenchyma, is a dynamic process. Newer cellular and biochemical investigations clarified its various steps. The process begins with stimulation of cells to connective tissue formation and can be divided into (1) intracellular synthesis, (2) extracellular maturation, and (3) collagen breakdown. The turnover of the connective tissue in the liver is conspicuously increased in chronic hepatitis of any type, as indicated by an elevation of several cellular and metabolic parameters. They are particularly raised in chronic hepatitis and in alcoholic liver injury. Further development of these parameters in the future should facilitate the analysis of the dynamics of fibroplasia. The strongest stimuli for hepatic fibroplasia are hepatocellular necrosis and inflammation, but ethyl alcohol and steatosis are also stimulating, though to a lesser degree. This explains the particular elevation of the fibroplastic parameters in alcoholic hepatitis. It points, however, also to the possibility that cirrhosis might develop without significant hepatocellular necrosis and inflammation. Perihepatocellular, periductular, and septal fibrosis are the functionally most important localizations leading to additional hepatic injury. The initiation of these types of fibrosis by liver injury points to a vicious circle. Specific anti-fibroplastic therapy is still in infancy.


Inhibition of diacylglycerol:CDPcholine cholinephosphotransferase activity by dimethylaminoethyl p-chlorophenoxyacetate. Cholinephosphotransferase [EC 2.7.8.2] activity of rat liver microsomes, with 1,2-di-0-[3H]acyl glycerol or 1-0-hexadecanoyl [U-14C]ethanediol as substrate, was inhibited by N,N-dimethylaminoethyl p-chlorophenoxyacetate (centrophenoxine). Inhibition progressed in a linear fashion with increasing drug levels and was complete at 30 mM concentration. It appears that the microsomal enzyme was largely affected by the drug itself because the hydrolysis products of centrophenoxine, viz., N,N-dimethylaminoethanol and p-chlorophenoxyacetic acid, were less inhibitory.


The circadian cycles of plasma corticosterone and adrenal cholesteryl esters in the normal and EFA-deficient female rat. In female rats subjected to a 12 hr light-12 hr darkness schedule and fed a semipurified diet containing 10% corn oil, plasma corticosterone concentration showed a monophasic circadian cycle with minimum and maximum concentrations at the start of the light and dark periods, respectively. Adrenal total cholesteryl ester concentration was inversely related to plasma corticosterone, as were those of several of the individual esters; changes in cholesteryl ester concentration appeared to follow rather than precede changes in plasma corticosterone. There was preferential depletion of the cholesteryl esters of 18:1, 18:2omega6, and 20:4omega6 during glucocorticoid secretion. [ EFA, essential fatty acid (s);X:YomegaZ, fatty acid with X carbon atoms and Y olefinic bonds with the terminal double bond Z carbon atoms from the methyl group.] In female rats fed hydrogenated coconut oil (EFA-deficient), a monophasic cycle for plasma corticosterone was also observed, but the peak was much broader than that recorded for rats fed corn oil, although minima and maxima occurred at similar times for the two groups. No significant cycle of adrenal total cholesteryl esters was evident in the deficient rats, but the 20:3omega9 and 22:3omega9 esters did decrease significantly during the period of high plasma corticosterone concentration. Preferential net decreases in adrenal cholesteryl esters during corticosteroidogenesis were more apparent in normal than in EFA-deficient rats.


Metabolism of apolipoprotein A-I in healthy young adults. The metabolism of 125I-labeled apolipoprotein A-I bound to high-density lipoproteins by an in vitro transfer procedure was studied in 10 healthy young adults (5 males and 5 females). Both sexes handled the labeled apolipoprotein similarly, and no statistically significant differences were found in the derived kinetic data. The mean (+/- 1 SD) plasma apolipoprotein A-I concentrations (males, 105 +/- 19 mg/dl; females, 111 +/- 13.8 mg/dl) and half-lives (males, 4.46 +/- 0.45 days; females, 4.64 +/- 0.70 days) were similar, as were the fractional rates of catabolism (FCR) of the apoprotein derived from the above data (FCR in males, 27% of intravascular pool/day; FCR in females, 25% of intravascular pool/day). The absolute catabolic rate of the apoprotein, equivalent under steady-state conditions to the synthetic rate, was 12.1 +/- 1.6 mg/kg/day in males and 11.9 +/- 2.4 mg/kg/day in females.


Regulation of aflatoxin biosynthesis. 1 Comparative study of mycelial composition and glycolysis in aflatoxigen and nonaflatoxigenic strains. A comparative biochemical study of an aflatoxigenic strain Aspergillus parasiticus NRRL 3240 and a nonaflatoxigenic strain A. flavus NRRL 3237 was carried out in order to have a better idea of regulation of aflatoxin biosynthesis. The results obtained revealed continuous primary metabolic activity (protein synthesis) in the nonaflatoxigenic strain while the aflatoxigenic stain showed inhibition of protein and nucleic acid synthesis. The aflatoxigenic strain showed higher levels of oxygen uptake, RNA, NAD, FMN and activities of glycolytic enzymes. Furthermore, it had lower of lipids and reduced activity of glucose-6-phosphate dehydrogenase, which is a source for NADPH. The differences observed have been discussed in relation to aflatoxin biosynthesis and its regulation.


Oxygen toxicity: comparative sensitivities of membrane transport, bioenergetics and synthesis in Escherichia coli. The oxygen sensitivities of basic cell functions were compared to evaluate their significance as potential causes of the reversible growth inhibition produced in Escherichia coli by exposure to hyperbaric oxygen. Growth and net incorporation of radioactive glucose into cell structure, and specifically in to protein, were completely inhibited in approximately 1/20 of a generation by a gas phase containing 4.2 atmospheres of oxygen. The inhibition occured before there was significant decrement in cellular glucose transport, respiration, or intracellular concentration of adenosine triphosphate. The data indicate that fundamental steps leading to protein biosynthesis from glucose should be examined in the search for specific primary sites of oxygen toxicity.


[Pancreatitis after renal transplantation (author's transl)]. A renal transplant patient in whom acute pancreatitis developed 2 1/2 years after surgery is presented. Pancreatisis was accompanied by hyperlipaemia, diabetes mellitus and acute renal failure possibly due to acute tubular necrosis. Pancreatic abscesses necessitated subtotal pancreatectomy 2 months later. Because of generalized tuberculosis finally the patient succumbed 6 weeks thereafter. As aetiological factors cytomegalovirus disease, disorders in lipid metabolism and immunosuppressive therapy must be discussed. It is concluded that prophylactic measures as well as early diagnosis, intensive care and therapy are necessary for reducing the high risks of pancreatis in renal transplant recipients.


Is the polio virus responsible for late central nervous system tumors? A hypothesis is advanced concerning the relationship of poliomyelitis viruses and late developing neoplasia in the central nervous system. The literature is surveyed, and several experimental approaches based upon known evidence of viral and neoplastic behaviour are advanced. The present status of world polio incidence is discussed as it applies to this possible relationship.


Regulation of the deo operon in Escherichia coli: the double negative control of the deo operon by the cytR and deoR repressors in a DNA directed in vitro system. The synthesis of the four enzymes of the deo operon in Escherichia coli is known from in vivo experiments to be subject to a double negative control, exerted by the products of the cytR and deoR genes. A DNA-directed in vitro protein synthesizing system makes the deo enzymes (exemplified by thymidine phosphorylase) in agreement with in vivo results. Enzyme synthesis is stimulated by cyclic AMP and repressed by the cytR and deoR gene products. Repression by the cytR repressor is reversed by cytidine or adenosine in the presence of cyclic AMP, while repression by the deoR repressor is reversed by deoxyribose-5-phosphate. Assays for the presence of the cytR and deoR repressors were established by use of S-30 extracts prepared from the regulatory mutants. Dissociation constants for repressor-operator binding as well as for repressor-inducer interactions have been estimated from the results.


A yeast mutant with glucose-resistant formation of mitochondrial enzymes. Yeast mutants with glucose-insensitive formation of mitochondrial enzymes were isolated starting with a strain completely lacking alcohol dehydrogenase activity. The mutations could uniquely be attributed to a single nuclear gene, designated CCR80. They were largely dominant. Glucose-resistant enzyme formation was most prominent with regard to mitochondrial enzymes succinate dehydrogenase and NADH: cytochrome c oxidoreductase. The effect of CCR80r mutations was rather small but significant on the gluconeogenetic enzymes isocitrate lyase, malate synthase and fructose-1,6-bisphosphatase and on invertase synthesis. The repressive effect of maltose in CCR80r mutants was also reduced showing that glucose-resistance is not caused by a mere hexose uptake defect. This regulatory disorders were not accompanied by reduced levels of glycolytic enzymes or drastically altered levels of glycolytic intermediates. Aerobic fermentation of glucose was almost completely inhibited in the mutants; anaerobic glucose degradation was reduced but not completely abolished. Therefore, the mutants appear to be altered in the regulation of glycolysis. A largely glucose-resistant synthesis of respiratory enzymes is obviously a corollary of this alteration.


[Effect of oxidizing and reducing agents on the rate of water proton relaxation in the chloroplasts of beans and particles of photosystem 2]. The change of longitudinal proton relaxation rate, 1/T1, in the native and shocked chloroplasts, PS2 chloroplasts' particles and exogenous complexes Mn2+--PPCI (pigment--protein complex of PSI), Mn2+--PPCII in the presence of the PS2 donors and acceptors and at different pH change were studied. It has been shown that 1/T1 increases in chloroplasts and PS2 particles in the presence of the PS2 donors and decreases in the presence of the acceptors in contrast to untreated preparations. The 1/T1 change of exogenous complexes is not specific. The results suggest that the manganese pool of the chloroplasts PS2 may be the main factor responsible for the chloroplasts' proton relaxation rate. The change of the manganese valency in the presence of donors and of acceptors may result in changing 1/T1.


[Effect of low-molecular nuclear RNA on RNA synthesis in isolated nuclei]. An attempt was made to elucidate possible participation of low molecular weight nuclear RNA's (LMWN RNA's) in the transcription process. For this purpose, we studied the effect of individual fractions of LMWN RNA's, isolated by polyacrylamide gel electrophoresis, on the endogenous RNA synthesis in isolated nuclei. We have found no influence of LMWN RNA's on the incorporation of labeled precursors in the acid-insoluble material under the conditions when RNA polymerase I is predominantly active. The results obtained thus indicate that LMWN RNA's do not participate in the regulation of 45S pre-rRNA synthesis and they do not belong to limiting factors in pre-rRNA synthesis.


Cellular reactivity to altered glomerular basement membrane in glomerulonephritis. Glomerular basement membrane may be altered during glomerulonephritis, exposing antigens that are recognized as foreign. Immunochemical studies suggest that removal of peripheral glycopeptides from the basement membrane with glycosidase mimics this pathogenetic event. To examine these hypotheses, we studied 24 patients with biopsy-proved glomerulonephritis by means of the lymphocyte-blast-transformation assay. Three preparations of normal glomerular basement membrane were used: two mimicked the native state for the peripheral glycopeptides, and one was altered by glycosidases. Results showed minimal differences in responses to native glomerular basement-membrane preparations among patients with glomerulonephritis and control groups. However, patients with glomerulonephritis had a significant blastogenic response to the glycosidase-treated glomerular basement membrane as compared to patients with nonglomerular renal disease and normal controls (P less than 0.0005). These studies suggest that cellular reactivity to altered glomerular basement-membrane antigens can be detected in certain forms of progressive glomerulonephritis.


Lymphoid changes suggesting escalation of immune response into malignant lymphoma. The conditions in which the microscopic changes of the lymphoid organs, especially lymph nodes, render difficult the histologic decision of immune response or malignant lymphoma, are reviewed. The possibility that in certain circumstances, prolonged, pathologic immune response escalate to malignant lymphomas, as well as the new classifications of these diseases, in terms of this conception are discussed.


Behavior of mitochondria from normal and tumor cells in isopycnic sucrose gradients. Rat and mouse liver mitochondria, when centrifuged in a sucrose density gradient (25--50%, w/w), showed the presence of heavy (H) and light (L) subfractions with buoyant densities 1.185 and 1.170--1.165 g/ml, respectively. Mild treatment with digitonin or EDTA (30 mM) shifted H-subfraction of mitochondria into the lighter zone of the gradient and as a result of this the mitochondria were distributed as a homogenous band with buoyant density 1.170--1.165 g/ml. Mitochondria isolated from both MD hepatoma and Zajdela rat hepatoma were characterized by a homogenous banding with buoyant density 1.160--1.165 g/ml. Regarding to this, the content and patterns of polyribosomes bound to outer membranes of mouse tumor mitochondria were studied. Analysis of polyribosomes as well as the results of RNA polyacrylamide gel electrophoresis indicate that the content of these polyribosomes in tumor mitochondria is less than that in normal liver ones. However, the decrease of cancer cell membrane-bound polyribosomes cannot account for the differences in buoyant densities of mitochondria from normal and tumor tissues.


The effect of psychotropic drugs on the TSH-response to thyroliberin (TRH). The effect of chlorpromazine (CPZ), thioridazine (TR), L-dopa, fusaric acid (FA), pimozide (PZ) and propranolol (P) in the i.v. thyrotropin-releasing hormone (TRH) stimulation test was studied in 75 subjects. It was found that chronic treatment with CPZ, TR and L-dopa inhibits the response to TRH. FA may to some extent counteract the effect of L-dopa. The results indicate that the response of the pituitary to TRH as regards the secretion of TSH is mainly regulated by norepinephrinergic alpha-receptors.


Growth hormone and ACTH in the pituitary of normal and anencephalic human fetuses: immunocytochemical evidence for hypothalamic influences during development. The influence of the hypothalamus on the development of the anterior pituitary cells was studied in normal and anencaphalic human fetuses. An immunocytological method was applied to study the quantitative evolution of the size of the cells during gestation. The somatotropic and corticotropic cells appeared at the end of the 2nd month of gestation. After the 3rd month of gestation their normal development required the presence of brain. The results suggest that hypothalamic factors are needed for the normal development of cells containing these pituitary hormones.


Thyrotropin-releasing hormone does not alter the release of melanocyte-stimulating hormone or adrenocorticotropic hormone from the rat pars intermedia. Thyrotropin-releasing hormone (TRH) has been reported to stimulate the release of melanocyte-stimulating hormone (MSH) from the pars intermedia (PI) of Rana esculenta. To test its effect on the rat PI, acutely dispersed rat PI cells, as well as whole nervosa-intermedias (NI), were incubated with synthetic TRH, from 10(--10)--10(--4)M. TRH did not alter the release of either MSH or adrenocorticotropic hormone (ACTH).


Computed tomographic evaluation of therapeutically induced changes in primary and secondary brain tumors. A prospective study was initiated to evaluate computed tomography as a method for monitoring therapeutically induced changes in brain tumors. Early postoperative scans may be misleading in the evaluation of residual tumor because of trauma to the blood-brain barrier during operation. The size of the dominant mass (17/25), enhancement (11/25), edema (11/25), and ventricular distortion (14/25) were decreased in many patients after radiation therapy. Occasional tumors increased in size during radiation therapy (3/25). Enlargement of the lateral ventricles developed in 9 of 25 patients. Computed tomography offers definite advantages over nuclide brain scans, angiography and other diagnostic studies for evaluating therapeutically induced changes in brain tumors.


Malignant fibrohistocytoma of the maxilla presenting with endodotically involved teeth. It is well established that a periapical lesion may be the first presenting form of more serious disease. A case of malignant fibrohistocytoma has been presented. It is of special interest because of the rarity of the disease, especially in the jaws, and because it presented as periapical disease associated with nonvital teeth. The case emphasizes the value of routine histopathologic examination of tissue and the need for maintaining a high "index of suspicion" when dealing with clinically atypical behavior. The value of consultation with an oral pathologist has also been stressed.


The role of antibiotics, immunizations, and adenoviruses in pertussis. Sixty-five patients with pertussis were identified by a clinical criterion, and Bordetella pertussis was isolated from 75% of these patients or their symptomatic household contacts. Negative nasopharyngeal cultures were usually associated with either a history of antibiotic therapy with erythromycin or tetracycline (two of three patients), two or more diphtheria and tetanus toxoids with pertussis (DTP) vaccines (six of eight patients), or both (two of three patients). Erythromycin therapy resulted in the elimination of B. pertussis from the nasopharynx in 2 to 7 days (mean, 3.6 days) compared with 7 to 17 or more days (mean, greater than 12 days) in patients treated with no antibiotics, but had no effect on the duration or severity of illness as judged by length of hospitalization. Adenoviruses were recovered from five of 44 patients cultured. Four of these isolates were from throat swabs obtained early in the illness and the remaining isolate was from one of 33 repeated viral cultures obtained two to three weeks later; B. pertussis was also isolated from these five patients. Paired serum samples were obtained from only two of these patients. Neither demonstrated a fourfold rise in adenoviral complement-fixing antibodies. Therefore, in these patients, adenoviral isolation may have been secondary to reactivation of a latent viral infection by infection with B. pertussis.


Adenovirus meningoencephalitis.

Adenovirus meningoencephalitis. Central nervous system manifestations in association with adenovirus infections have rarely been documented. In this report, four children, ages 4, 5, 6, and 10 years, with adenoviral meningoencephalitis are described. Adenoviruses were recovered from CSF specimens in all four patients.


Inability of myoglobin to increase in dystrophic skeletal muscle during daily exercise. An exercise program consisting of 80-min daily runs on a treadmill was performed by normal and dystrophic hamsters. Subgroups were sacrificed at various times during the 45-day program. Daily exercise resulted in a significant increase in the myoglobin concentration of gastrocnemius muscles in normal animals but not in dystrophic animals. In the exercise groups of hamsters, there were significant increases in the concentration of cytochrome c, a marker for respiratory capacity, in the gastrocnemius of both normal and dystrophic hamsters.


The sensitivity of pineal gland beta-receptors appears to be dependent upon calcium ions. Rat pineals were used in vitro to demonstrate that calcium antagonists induce a hyposensitive response to stimulation of serotonin N-acetyl transferase (NAT) by the beta-agonist isoproterenol or by dibutyryl cyclic AMP. Mn2+, Co2+ and La3+ at 10(-3)M all significantly reduce the effects of isoproterenol stimulation. In addition, the two enantiomers of D600 also inhibit NAT induction, the D-form slightly more effectively than the L-form. Interestingly, La3+ is also able to inhibit NAT after DBcAMP, indicating that Ca2+ can also control enzyme induction at an intracellular site beyond the beta-receptor.


A study of the interaction of catecholamines and antidiuretic hormone on water permeability and the cyclic AMP system in isolated papillae of the rat. The diffusional water permeability of collecting ducts in vitro and the cyclic A.M.P. content of isolated papillae were measured after exposure to different concentrations of antidiuretic hormone, isoproterenol and noradrenalin. Antidiuretic hormone 25 mu units/ml. caused a 25% increase in diffusional water permeability. This response was not affected by isoproterenol (10(-6) M) or noradrenalin (2 x 10(-6) M). Antidiuretic hormone 100 mu unit ml-1 caused a 50% increase in diffusional water permeability which likewise was not altered by isoproterenol or noradrenalin. Isoproterenol (10(-6) M) and noradrenalin (2 x 10(-6) M) had no significant effect on basal levels of diffusional water permeability. Isoproterenol had no significant effect on the tissue concentration of cyclic A.M.P. concentration induced by antidiuretic hormone. Noradrenalin (2 x 10(-6) and 10(-4)) had no significant effect on basal cyclic A.M.P. concentration. However, noradrenalin inhibited the stimulation of cyclic A.M.P. induced by antidiuretic hormone. This effect was inhibited by phentolamine. This study suggests that catecholamines do not alter water handling by a direct action on the water permeability of the kidney but probably exert their action through an effect of A.D.H. release.


The prophylactic use of vitamin D3 in parturient paresis in the cow in a practice district in eastern Norway. A retrospective investigation. The prophylactic effect in parturient paresis of an intramuscular injection of 10 million i.u. vitamin D3 one week prior to the expected calving date, was investigated in 84 cows which had previously suffered from parturient paresis. Clinical observations in connection with the development of parturient paresis after injection of vitamin D3 were compared with corresponding observations made on cows within the same population at the previous calving when no vitamin D3 was administered. The incidence of clinical parturient paresis in cows given vitamin D3 was 44.3 per cent. There were no significant differences in the incidence of parturient paresis in cows injected with vitamin D3 during the periods less than 2, 2--4, 5--8 and greater than 8 days before parturition. Average plasma calcium levels (mg/100 ml) before first calcium treatment, results of treatment, and the incidence of retained placenta in vitamin D3 injected cows did not differ significantly from corresponding data for cows with clinical parturient paresis which had not been given vitamin D3 before calving. It is concluded that vitamin D3 does not have any prophylactic effect in parturient paresis in cows in Eastern Norway.


Gene mapping and gene enrichment by the avidin-biotin interaction: use of cytochrome-c as a polyamine bridge. A modification of previously described methods of electron microscopic gene mapping and of gene enrichment based on the avidin-biotin interaction is presented. The modification consists of coupling cytochrome-c instead of pentane diamine to the oxidized 2', 3' terminus of an RNA by Schiff base formation and BH-4 reduction. The RNA-cytochrome-c conjugate is purified by a simple chromatographic procedure; several biotins are attached to the cytochrome moiety by acylation. The extended arm between biotin and RNA gives efficient electron microscopic gene mapping of DNA:RNA-biotin hybrids with avidin-ferritin and avidin-polymethacylate sphere labels and efficient gene enrichment by buoyant banding of DNA:RNA-biotin:avidin-spheres in CsCl. A 1400 fold enrichment (thus, 25% pure) and a 90% yield of long Drosophila DNA strands with 5S RNA genes is achieved.


Base-specific reactions useful for DNA sequencing: methylene blue--sensitized photooxidation of guanine and osmium tetraoxide modification of thymine. Exposure of DNA to methylene blue and visible or ultraviolet light causes guanine-specific modification, and subsequent treatment with piperidine leads to chain cleavage at each guanine residue. Treatment of DNA with osmium tetraoxide in dilute pyridine leads to thymidine-specific modification, and subsequent treatment with piperidine leads to chain cleavage at the modified thymidine residues. Both reactions can be used in conjunction with other base specific modifications described by Maxam and Gilbert (1) for the determination of the nucleotide sequence in DNA.


Interaction of the DNA untwisting enzyme with the SV40 nucleoprotein complex. The SV40 nucleoprotein complex which was isolated from infected CV-1 cells did not possess an active DNA untwisting enzyme. The superhelix density of the DNA in the chromatin complex was unchanged after treatment with purified rat liver DNA untwisting enzyme. However, in the presence of ethidium bromide (1 microgram/ml) the superhelix density was changed. Moreover, the nicked intermediate in the DNA untwisting reaction could be detected using the chromatin DNA as a substrate. These results show that the DNA in the SV40 chromatin which is accessible to the DNA untwisting enzyme is under no topological strain.


Cytomegalovirus infection and the Guillain-Barré syndrome. A fatal case is described of the Guillain-Barré syndrome due to cytomegalovirus infection, which was associated with hepatitis, myocarditis and viral pneumonia. Previous cases, which have usually run a benign course, are reviewed. Attention is drawn to the possible adverse effect of steroid therapy.


[Cytochrome oxidase activity in diabetes mellitus]. The results of study of the activity of cytochromeoxidase in the blood formed elements in 84 patients suffering from diabetes mellitus before and after the treatment are presented. In patients with the primarily detected disease the activity of the enzyme was reduced, and in its prolonged course--it was elevated. A tendency to normalization of the enzyme activity was noted as a result of treatment in all the forms of the course of the disease; however, the interstitial metabolism failed to become restored completely.


[Various mechanisms of disorders of the hypothalamo-hypophyseal-adrenal system in Cushing's syndrome]. The authors present the results of study of corticotropine and cortizol secretion under the effect of stress caused by insulin hypoglycemia, of sensitivity of the adrenal glands to the administration of exogenous synthetic corticotropine, and of the 24-hour rhythm of the corticotropine and cortizol secretion in 26 healthy persons and in 57 patients suffering from Itsenko-Cushing's disease, including 25 after a course of treatment with chloditan, an inhibitor of the adrenal cortex function. Blood corticotropine content was determined by the radioimmunological method, and of cortizol--by competitive binding. There was revealed no significant increase of corticotropine and cortizol secretion in response to insulin hypoglycemia in Itsenko-Cushing's disease, this pointing to disturbance of the hypothalamo-hypophysio-adrenal system at the hypothalamic level. No qualitative differences were found in response of the adrenal cortex to corticotropine, this indicating the absence of changes of its sensitivity to this hormone. The 24-hour rhythm of corticotropine and cortizol secretion in this disease is inverted. At the immediate periods after chloditan treatment no normalization of the response of the adrenal cortex to insulin hypoglycemia, and of the 24-hour rhythm of corticotropine and cortizol secretion, and also of the changes in the adrenal cortex sensitivity to corticotropine was seen.


[Stromal thecomatosis of the ovaries associated with the syndrome of mixed ovarian dysgenesis]. On the basis of analysis of a case of a combination of mixed dysgenesia of the ovaries with stromal thecomatosis and the clinical picture of the virile syndrome a conclusion was drawn that a) hypertestosteronemia was in such cases connected with the presence in the dysgenetic ovary and the connective tissue band of epitheliod cells forming the foci of stromal thecomatosis, and b) that the appearance of stromal thecomatosis failed to depend on the presence of the follicular apparatus; apparently cells of the interstitial tissue of the cortical layer of the ovaries serve as its source.


[Disorders of lipid content of blood in diabetes mellitus]. The authors present the results of prospective study of the blood serum lipids (total cholesterol, triglycerides, lipoproteid fractions) in 132 patients with diabetes mellitus aged from 35 to 60 years and in 47 healthy persons of the same age (control group). The majority of the patients examined (60.6%) showed hyperlipidemia; types II and IV were most frequent. An increase in the content of the probeta-lipoproteid fraction and hypertriglyceridemia were mostly encountered in patients with macroangiopathies, particularly in the presence of adiposity. There was revealed no significant relationship between the frequency of hyperlipidemia and the severity of diabetes mellitus. However, the most distinct lipid disturbances were found in the patients with the disease of moderate severity. This was apparently connected with the prevalence in this group of persons with an excessive weight. As revealed, the incidence of hyperlipidemia rose on account of increased level of both total cholesterol and of triglycerides (with a tendency to their greater content in women).


[Activity of glutathione reductase in the adrenal glands and the liver of dogs after administration O,P-DDD, perthane and ACTH]. A study was made of the inhibitors of the adrenal gland function and ACTH on the activity of NADP-specific glutathionreductase in the adrenal glands and the liver of dogs. After a 10-day feeding to dogs of o,p'-dichlordiphenyldichlorethane (o,p'-DDD) and p,p'-diethyldiphenyl dichlorethane (p,p'-pertane) in a dose of 50 mg/kg there was observed a marked activation of glutathion reductase in the adrenal glands. Elevation of the activity of the enzyme was revealed as soon as 24 hours after the administration of o,p'-DDD. The inhibitors under study failed to influence the activity of the enzyme in the liver of dogs, this indicating a specific action of o,p'-DDD and pertane on the adrenal gland cortex. Stimulation of glucocorticoid biosynthesis under the effect of ACTH administered once in a dose of 25 Units/kg was not accompanied by the changes in the activity of glutathione reductase in the adrenal glands and the liver of dogs. Prolonged ACTH administration (4 Units/kg) caused a significant reduction of the enzyme activity calculated as per 1 mg of protein in the adrenal gland homogenate.


Blockade of neuromuscular transmission by enzymatically active and inactive beta-bungarotoxin. beta-Bungarotoxins have been shown to be presynaptic blockers of neuromuscular transmission. This paper reports experiments using the most positively charged beta-bungarotoxin that elutes from a CM-Sephadex C-25 column. The toxin is shown to be a single polypeptide with a molecular weight of approximately 11,000 and has phospholipase A2 activity. The application of the enzymatically active toxin to the frog sciatic nerve-sartorius muscle preparation results in an initial decrease in the average endplate potential amplitude followed by a temporary rebound in endplate potential amplitude, and finally a complete inhibition of endplate potentials. Similarly, minature endplate potential frequency is initially reduced upon toxin application but then increases dramatically. After the phospholipase A2  of the toxin is inactivated, treatment with the toxin results in only the initial decrease in transmitter release. There results suggest that this beta-bungarotoxin acts in two functionally separate steps: (i) by binding to a specific presynaptic site possibly associated with calcium entry, and (ii) by perturbing the presynaptic membrane by its enzyme action, which results in an increase and then a failure in transmitter release.


An RNA transcribed from DNA at the origin of phage fd single strand to replicative form conversion. Phage fd DNA complexed with DNA binding protein I was used by Escherichia coli RNA polymerase (nucleoside triphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) to synthesize an RNA at the origin of single strand to double strand replication. The isolated ori-RNA gave a simple fingerprint after nucleolytic digestion and has a length of about 30 nucleotides. The characterization of the oligonucleotides from the nuclease digest and the extension of the ori-RNA with DNA polymerase I and subsequent restriction of the DNA gave its exact localization in the fd genome, and its total sequence was deduced from the known DNA sequence in this region.


Characterization of pro-opiocortin, a precursor to opioid peptides and corticotropin. The high molecular weight (approximately 30,000) precursor to opioid activity (pro-opiocortin) previously detected in extracts of rat pituitary was digested with trypsin and the resulting peptide mixture was resolved by high-performance reverse-phase chromatography. A peak of opioid activity was eluted at the position of the nonapeptide beta-LPH (61-69), which was also the same fragment obtained by trypsin digestion of betas-lipotropin or beta-endorphin. This identified the protein as a precursor to the endorphins and Met-enkephalin. No activity was detected in the position corresponding to the Leu5 analog of betas-LPH (61-69), thus ruling out the possibility of a beta-lipotropin-like precursor to Leu-enkephalin in pituitary extracts. Pro-opiocortin and beta-lipotropin are present in rat pituitary extracts in comparable amounts, approximately 10 pmol/mg of tissue.


In vitro synthesis of vesicular stomatitis virus membrane glycoprotein and insertion into membranes. Translation in vitro of the mRNA coding for the vesicular stomatitis virus membrane glycoprotein G in a membrane-free ribosomal extract from HeLa cells allowed the synthesis of only the unglycosylated protein G1 (molecular weight, 63,000). Addition of stripped crude microsomal membranes from HeLa cells resulted in the conversion of G1 to the glycosylated protein G2 (molecular weight, 67,000). The G2 protein synthesized by the reconstructed microsomal membrane/ribosome system was found to be segregated inside the microsomal membrane vesicles and was thus protected from the proteolytic action of trypsin and chymotrypsin. Stripped membranes were required at an early stage of protein synthesis for the synthesized protein to be inserted into the membrane vesicles and to be glycosilated. The segregated protein G2, however, was not completely protected from proteolytic digestion, showing that a portion of the polypeptide chain of about 3000 daltons was present on the cytoplasmic side of the membrane vesicle. Our data thus suggest that, unlike the secretory proteins, the membrane glycoproteins are not completely discharged across the microsomal membranes.


Triphosphate residues at the 5' ends of rRNA precursor and 5S RNA from Dictyostelium discoideum. We present here direct evidence for the preservation of a transcriptional initiation sequence in a eukaryotic rRNA precursor: the 5'-end group for precursor to 17S rRNA (p17S RNA) from Dictyostelium discoideum is identified as the triphosphate residue pppA-. We also show that mature 5S RNA form Dictyostelium bears a different triphosphate residue, pppG-. In contrast, we find no evidence for more than one phosphate at the 5' end of the 25S rRNA precursor (p25S RNA). These observations indicate that synthesis of the large ribosomal RNAs of Dictyostelium begins with the 5'-terminal sequence of the p17S RNA, and that 5S RNA transcription must be initiated independently, despite the close association of the 5S and rRNA coding segments.


Direct fluorescent assay of urokinase and plasminogen activators of normal and malignant cells: kinetics and inhibitor profiles. A direct rate assay for plasminogen activator has been developed using a synthetic fluorogenic peptide substrate, 7-(N-Cbz-glycylglycylargininamido)-4-methylcoumarin trifluoroacetate. The assay correlates well with the standard 125I-labeled fibrin plate assay using highly purified urokinase, culture fluids from WI-38, Chinese hamster vary or HeLa cells, or Rous sarcoma virus-transformed chick fibroblasts as the source of plasminogen activator. The assay is sensitive, rapid, and linear throughout a wide range of enzyme concentrations. With this substrate it is possible to determine inhibitor profiles for the various plasminogen activators, independently of the interfering potential of plasmin. All of the enzymes tested are inhibited by leupeptin and antipain but not by the related aldehydes, elastatinal and chymostatin. The macromolecular inhibitors soybean trypsin inhibitor and trasylol have little or no effect on the plasminogen activators tested. This substrate should be useful for the study of the effect of various agents on functional changes in cells secreting this enzyme and also should allow kinetic measurements of potential inhibitors.


Oligosaccharide chains are trimmed during synthesis of the envelope glycoprotein of vesicular stomatitis virus. The biosynthesis and maturation of the oligosaccharide moieties of the envelope glycoprotein of vesicular stomatitis virus were investigated in virus-infected HeLa and BHK21 cells after pulse labeling with [2-3H]mannose. Two major forms of the virus glycoprotein were detected by polyacrylamide gel electrophoresis, which appear to correspond to the viral glycoprotein with either "precursor" or "mature" oligosaccharide chains. The precursor chains in both HeLa and BHK21 cells infected with vesicular stomatitis virus obtained after a 30-min pulse were large oligomannose structures containing approximately 7--9 mannose residues as estimated by gel filtration analysis. The size of the oligomannose structures initially transferred to the protein may have been even larger. Mature, virus-size oligosaccharide chains, which could be detected after a 20- to 30-min delay, contained only three mannose residues and, in addition, contained branch structures terminating in sialic acid. A precursor--product relationship of these two forms of oligosaccharide chains was demonstrated by pulse--chase labeling of virus-infected HeLa cells. These studies indicated that the large oligomannosyl core structures initially added to the glycoprotein were being "trimmed" by the removal of mannose residues prior to (and/or during) the addition of the branch chains terminating in sialic acid.


Alteration of the physicochemical properties of triphosphoinositide by nicotinic ligands. The concentrations of nicotinic drugs required to effect a 50% transfer of triphosphol[3H]isositide from an aqueous phase to a nonaqueous phase closely approximated their concentrations for both in vivo neuromuscular blocking activity and binding to purified nicotinic receptors, and correlated well (correlation coefficient = 0.95)) with their biological activities measured by other workers in an Electrophorus electroplax preparation. The triphospho[3H]inositide transfer induced by nicotinic ligands was dependent on the lipid concentration and was potentiated by Ca2+. The affinity constants of 45Ca2+ for triphosphoinositide were similar to those for the purified nicotinic receptor. These and other findings suggest the possibility that triphosphoinositide (phosphatidylinositol bisphosphate) is a binding site of the nicotinic receptor.


Structure of a poly (adenosine diphosphoribose) monomer: 2'-(5"-hosphoribosyl)-5'-adenosine monophosphate. A preparation of poly(adenosine diphosphoribose) synthase obtained from pigeon liver nuclei has been used to make poly(adenosine diphosphoribose) with an average chain length of 20. Digestion of the purified poly(adenosine diphosphoribose) with snake venom phosphodiesterase (oligonucleate 5'-nucleotidohydrolase; EC 3.1.4.1) gave the monomer, 2'-(5"-phosphoribosyl)-5-AMP. After purification of the monomer on a Dowex-1 column, further digestion with alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum); EC 3.1.3.1] yielded the dephosphorylated product, 2'-ribosyl adenosine. Nuclear magnetic resonance spectra at 360 MHz of the 2'-ribosyl adenosine were obtained in [2H6]dimethylsulfoxide, which allows direct observation of the hydroxyl protons. These spectra show the absence of the adenosine 2'-hydroxyl proton, thus confirming the 2' position as the site of attachment of the ribose to the adenosine moiety. Comparison of the coupling constants and the chemical shifts of the ribose hydroxyl protons of 2'-ribosyl adenosine with the model compounds alpha- and beta-methylribofuranoside establishes an alpha (1" leads to 2') glycosidic linkage in the monomer. No evidence was found for heterogeneity in either the site of attachment or configuration of the linkage in the 2'-(5"-phosphoribosyl)-5'-AMP.


Ionophore A-23187- and thrombin-induced platelet aggregation: independence from cycloxygenase products. Stimulation of platelets labeled with [14C]-arachidonate by ionophore A23187 or thrombin produces rapid degradation of specific membrane phospholipids. This is also reflected by the release of [14C]archidonate, which is immediately transformed into products of the cycloxygenase and lipoxygenase enzyme systems, and by increased labeling of phosphatidic acid. Arachidonate metabolism can be effectively prevented by preincubation with indomethacin and eicosatetraynoic acid, but platelet aggregation induced by ionophore A23187 or trombin is not blocked under these conditions. Nevertheless, in the virtually total absence of metabolism of arachidonate, platelet aggregation still occurs concomitantly with phospholipid breakdown and with increased labeling of phosphatidic acid. Increased levels of cyclic AMP block both phospholipase activation and aggregation induced by ionophore A23187 and trombin. These data suggest that some early consequence of phospholipase activation, independent of a metabolic product of arachidonate but possibly related to the production of phosphatidic acid, may play a central, causative role in mediating platelet aggregation.


Erythrocyte membrane abnormalities in Duchenne muscular dystrophy monitored by saturation transfer electron paramagnetic resonance spectroscopy. Saturation transfer electron paramagnetic resonance and the spin label 2-(3-carboxypropyl)-4,4-dimethyl-2-tridecyl-3-oxazolidinyloxyl were used to study erythrocytes from patients with Duchenne muscular dystrophy or Becker syndrome and from age-matched normal boys. There were significant differences in the spectral intensities of erythrocytes from Duchenne patients when compared to controls. Spectral intensities increased with time in the former; no such change was observed in the latter. Saturation transfer electron paramagnetic resonance spectra of erythrocytes from patients with Becker syndrome were significantly different from those from Duchenne patients but were not significantly different from normals. These observations suggest the possible usefulness of these techniques in the differential diagnosis of Duchenne muscular dystrophy. Spin label concentration spectral studies suggest that the observed spectral differences between Duchenne patients and controls were due to differential spin exchange phenomena.


Cyclic AMP-dependent protein kinase mediates a cyclic AMP-stimulated decrease in ornithine and S-adenosylmethionine decarboxylase activities. Incubation of S49 lymphoma cells with N6,O2'-dibutyryl cyclic AMP (Bt2cAMP) decreases the activities of ornithine decarboxylase (L-ornithine carboxy-lyase; EC 4.1.1.17) and S-adenosylmethionine decarboxylase (S-adenosyl-L-methionine carboxy-lyase; EC 4.1.1.50), the two principal enzymes in the pathway of polyamine synthesis. This decrease is dose-dependent, commences after a 3-hr delay, virtually abolishes the assayable activities of the two enzymes, and is not associated with a soluble inhibitor of the enzyme activities. Studies in mutant S49 clones that have altered protein kinase indicate that cAMP-dependent protein kinase mediates the decreases in enzyme activities. The dose-response pattern for the cAMP-stimulated decrease in enzyme activities parallels the pattern for the cAMP-stimulated, cell cycle-specific (G1) growth arrest of S49 cells. The activity of ornithine decarboxylase decreases faster than Bt2cAMP arrests wild-type S49 cells and, similarly, release of cells from the cAMP-stimulated arrest in G1 increases the activity of ornithine decarboxylase faster than cells exit from G1. These findings contrast with reports that cAMP induces ornithine decarboxylase in other cell types and further suggest that passage of cells through cell cycle is required for maintaining the activities of ornithine and S-adenosylmethionine decarboxylases.


Chemotactic attraction of human fibroblasts to type I, II, and III collagens and collagen-derived peptides. The chemotactice response of human dermal fibroblasts of type I, II, and III human collagens and collagen-derived peptides was quantitated by an in vitro assay. All three native human collagens and constituent alpha chains can serve as chemoattractants for fibroblasts in vitro. When type I, II, and III collagens were digested by bacterial collagenase, the resulting peptides were also chemotactic. In addition, synthetic di- and tripeptides containing hydroxyproline were also chemotactic for fibroblasts. Since collagen is degraded and remodeled at sites of tissue injury and inflammation, these findings suggest that collagen and collagen-degradation peptides might function as chemotactic stimuli for fibroblasts in vivo and attract these cells to effect repair of damaged tissue.


Cotransfer of thymidine kinase and galactokinase genes by chromosome-mediated gene transfer. The Chinese hamster genes for thymidine kinase (ATP:thymidine 5'-phosphotransferase, EC 2.7.1.75) and galactokinase (ATP:D-galactose 1-phosphotransferase, EC 2.7.1.6) have been cotransferred to mouse cells by chromosome-mediated gene transfer. Hamster metaphase chromosomes were incubated with mouse B82 cells and 22 independent colonies were isolated in a selective medium. All of the 12 colonies analyzed expressed the donor form of thymidine kinase; the hamster form of galactokinase was also expressed in 2 of these colonies, indicating cotransfer with a frequency of about 20%. There was coordinate loss of both transferred genes from each colony when selection was applied for the loss of thymidine kinase alone. Comparison of the regional localization of these two linked genes with the frequency of cotransfer suggests that the transgenome is probably not larger than about 0.25% of the donor genome.


Antibody response of mice to chemically induced tumors. BALB/c mice immunized with syngeneic methylcholanthrene-induced fibrosarcomas did not have antibodies against the unique tumor-specific transplantation antigens, even though they were capable of rejecting a lethal dose of tumor cells. Endogenous murine leukemia virus antigens expressed by certain of the tumors did, however, elicit high titers of antibodies, accounting for serological crossreactions that occurred between those tumors.


Vesicular stomatitis virus glycoprotein is necessary for H-2-restricted lysis of infected cells by cytotoxic T lymphocytes. Vesicular stomatitis virus (VSV) elicited cytotoxic thymus-derived lymphocytes (CTLs) in mice of the BALB/c and three congenic strains (BALB.b, BALB.k, BALB.HTG). CTL lysis of VSV-infected fibroblasts from the four strains was restricted by the target cells' major histocompatibility complex (H-2). Target cells were also infected with two temperature-sensitive mutants of VSV, tsM and tsG in which, respectively, the viral matrix protein and glycoprotein are not expressed at 39 degrees (restrictive temperature) on the infected cell's surface membrane. At the restrictive temperature, cells infected with wild-type VSV or tsM were lysed by CTLs, but cells infected with tsG were not. The requirement for the glycoprotein on the target cell was also evident from the ability of antisera to the glycoprotein to block completely CTL lysis of VSV-infected cells.


Control of aldosterone secretion during sodium restriction: adrenal receptor regulation and increased adrenal sensitivity to angiotensin II. The mechanism of increased adrenal sensitivity to angiotensin II during the aldosterone response to sodium restriction was investigated in the rat. Sodium restriction for 36 hr markedly increased the aldosterone-stimulating effect of low-dose (1 ng/min) infusion of angiotensin II and caused enhanced binding of (125)I-labeled angiotensin II to the zona glomerulosa in vivo. Conversely, in vivo binding of (125)I-labeled angiotensin II was significantly decreased after 36 hr of high-sodium intake. In isolated glomerulosa cells, the increased binding of angiotensin II after sodium restriction was shown to result from a significant increase in receptor affinity (+80%) and a smaller increase in receptor concentration (+25%). The corresponding aldosterone responses in dispersed cells showed an increase in sensitivity to angiotensin II, commensurate with the increased receptor affinity. More prolonged sodium restriction (4 days) caused a further increase in angiotensin receptor concentration (+70%) and maximal aldosterone response (+50%), whereas the binding affinity of adrenal receptors and the sensitivity of the in vitro aldosterone response had returned to normal. During sodium loading for 36 hr and 4 days, the converse effects on adrenal angiotensin II receptors and aldosterone production were observed. Also, in contrast to the consistent increase in angiotensin II receptors in the adrenal glands of sodium-restricted animals, the angiotensin II binding capacity of uterine smooth muscle was decreased by 40% after 7 days of sodium restriction.The rapid regulation of receptor affinity and concentration during changes in sodium intake provides a basis for the dynamic modulation of aldosterone responses by dietary sodium content. During sodium restriction, the sequential changes in receptor affinity and concentration account for the enhanced binding and steroidogenic actions of angiotensin II in vivo and in vitro. These receptor changes, and the converse effects of sodium loading, serve as a local regulatory mechanism in the physiological control of adrenal sensitivity and aldosterone secretion. The opposite finding in smooth muscle-that sodium restriction decreases the concentration of angiotensin II receptors-is consistent with the divergent effects of changing sodium balance upon vascular and adrenal responses to angiotensin II.


Extrahypophysial distribution of corticotropin as a function of brain size. Determination by radioimmunoassay of corticotropin in the brains of rats, rabbits, dogs, monkeys, and human beings reveals that the dimensions within which the hormone is found is about the same for each of these species but that the anatomical regions in which the hormone is found depends on brain size. Corticotropin is widely distributed in the brain of rats but is found only in the hypothalamic region of the primate brain. The patterns of immunoreactivity observed after Sephadex gel filtration confirm that the molecular forms of corticotropin found in extrahypophysial regions are similar to those in the pituitary of each species. These findings suggest that the mammalian pituitary is the sole site of synthesis of the hormone. The observation of persistence of corticotropin in the brains of commerically hypophysectomized rats has been interpreted by others as suggesting diencephalic as well as pituitary origin for this peptide. However, our studies demonstrate that 8 weeks after hypophysectomy the rats we have received from commerical sources manifest stress-stimulated plasma corticotropin concentrations about 80% of that found in intact rats in spite of the fact that residual pituitary tissue was not found by visual inspection of the sella. Scrapings from the sella revealed a corticotropin content up to 5% that of the average rat pituitary.


Infection of mouse preimplantation embryos with simian virus 40 and polyoma virus. Mouse two-cell embryos, morulae, and blastocysts were killed when infected in vitro with simian virus 40 (SV40) at high multiplicities of infection. Polyoma virus was not deleterious for preimplantation embryos, even at a very high multiplicity of infection; however, the outgrowths of polyoma-infected blastocysts disintegrated after several days of culture. Indirect immunofluorescence tests revealed the presence of SV40 T and V antigens and polyoma virus V antigen in the nuclei of trophoblastic cells. Virus-specific antigens were not found in the nuclei of cells forming inner cell masses of blastocysts or in inner cell mass-derived cells in blastocyst out-growths. The appearance of SV40 T and V antigens in the nuclei was inhibited by alphaamanitin, a RNA polymerase II inhibitor. The amount of infectious virus recovered from cultures of morulae or blastocysts on subsequent days after infection with SV40 initially declined but later increased. These points of evidence indicate that some cells of early mouse embryos are permissive for the expression of early and late functions of SV40 genome and that susceptibility to infection with polyoma virus and/or permissiveness for the expression of polyoma virus late functions develop gradually between the two-cell and blastocyst stages. Electron microscope observations showed the presence of specific complexes of membranes and virions in the cytoplasm of trophoblastic cells. Single viral particles could be found in the nuclei and also in mitochondria.


Oral absorption and selective tissue localization of 4'-(9-acridinylamino)-methanesulfon-m-anisidide. The disposition of 4'-(9-acridinylamino)-methanesulfon-m-anisidide (AMSA), a new antitumor agent presently undergoing clinical evaluation, was studied in mice and rats following oral administration and compared to that observed following intravenous administration. The metabolic fate of  AMSA was the same with either intravenous or oral administration; however, the tissue distribution of AMSA differed significantly between the two routes of administration. Following absorption from the GI tract, AMSA was rapidly cleared from plasma by the liver and excreted in the bile as metabolites. Concentrations of AMSA in the liver were relatively high after oral administration and were sufficient to exert a cytotoxic effect on L1210 cells implanted at the site. The results indicate the use of AMSA orally to attain selective localization in the liver with decreased systemic exposure, which may prove useful against tumor metastases to the liver or primary hepatocellular carcinoma.