Cortical and subcortical projected foci in cats: inhibitory action of taurine.

The effects of local application of taurine and isethionic acid on the propagation of the epileptic activity to the mirror area in cats have been studied. Cortical and amygdaloid acute foci were induced by local administration of conjugated estrogens. Taurine proved to be effective in reducing and sometimes in abolishing the appearance of the epileptic propagated elements in the mirror area. When this agent was applied 30 minutes before the induction of the primary focus, the single spike transmission was reduced or prevented; however, the transmission of a seizure was not blocked. No changes in the transmitted phenomena were observed when isethionic acid was administered with the same technique as that used for taurine. The present study stresses the clear antiepileptic activity of taurine in this experimental model, ruling out the possibility of an unspecific interaction with the epileptogenic agents. Moreover, it is suggested that the deamination of taurine is not important for its antiepileptic action.


Nuclear bodies in Creutzfeldt-Jakob and Alzheimer's diseases.

Nuclear bodies types I, II, III, and IV, as described by Bouteille, Kalifat, and Delarue, have been found in one case of Creutzfeldt-Jakob disease and one case of Alzheimer's disease. The relationship of nuclear bodies to viral disease in the central nervous system is not clear.


[Muscle cell structure in polymyositis].

Electron microscopic investigations of muscles from patients with chronic polymyositis demonstrated presence of cytoplasmatic, tubular structures and extracellular virus-like particles. Their size, morphology and location suggest multiplication of viruses from the group of parainfluenza viruses within the muscle cell in "infected" cultures of human fetal muscles. Beside viruses belonging to the parainfluenza group presence of viruses from the herpes group was demonstrated. The latter belong to latent viruses.


[Case of pseudopseudohypoparathyroidism].

A case of pseudo-pseudo-parathyroidism, in which normal blood chemistry data were accompanied by marked shortening of the 4th metacarpus and metatarsus, and a suggestion of syndactyly, is reported.


Electron microscopy of Cowdria ruminantium infected reticulo-endothelial cells of the mammalian host.

An electron microscopic study of structures suspected to represent a possible developmental cycle of Cowdria ruminantium in reticulo-endothelial cells of mice and ruminants is reported. After infection dense bodies increase in size and undergo division to form fragmented dense bodies. These in turn apparently sub-divide and become organized to give rise to mature "organisms". In none of these structures do limiting membranes separate the parasitic inclusions from the host cell cytoplasm. Present observations suggest that growth of the organism in reticulo-endothelial cells differs from that of chlamydial and rickettsial agents and somewhat resembles the replication of some viruses. Developmental stages observed after infection of ruminants with the Ball 3 strain of the heartwater agent are indistinguishable from those seen with the mouse adapted strain. These observations support the hypothesis that C. ruminantium released from reticulo-endothelial cells subsequently penetrates endothelial cells where further multiplication by binary fission occurs.


Effect of Clostridium perfringens epsilon toxin on the blood brain barrier of mice.

It was shown that Clostridium perfringens epsilon toxin has the effect of allowing the passage of 125I polyvinyl-pyrrolidone and 125I human serum albumin into mouse brain. These substances did not enter the brains of normal control mice. The passage of albumin into the brains of mice poisoned with epsilon toxin was extremely rapid. When large doses of toxin (+/-4 000 MLD) were given death ensued within 2-3 min at which stage 1,5% of the injected albumin had already entered the brain. In cases where smaller doses were given and the time interval between injection and death was longer the figure was increased to 2-2 1/2% of the injected plasma albumin.


Occurrence of juvenile angiofibroma in middle age.

A case of juvenile angiofibroma occurring in a male patient of 49 years is described. Histopathology and oncogenesis are discussed.


Asymmetric adsorption by quartz: a model for the prebiotic origin of optical activity.

One mechanism previously proposed for the abiotic accumulation of molecules of one chirality in nature is asymmetric adsorption on the chiral surfaces of optically active quartz crystals. Earlier literature in this field is reviewed, with the conclusion that previous investigations of this phenomenon, using optical rotation criteria, have afforded ambiguous results. We now have studied the adsorption of radioactive D- and L-alanine on powdered d- and l-quartz, using change in radioactivity level as a criterion for both gross and differential adsorption. d-Quartz preferentially adsorbed D-alanine from anhydrous dimethyl-formamide solution, and l-quartz L-alanine. The differential adsorption varied between 1.0 and 1.8%. The implications of these observations are discussed from the viewpoint of early chemical evolution and the origin of optically active organic compounds in nature.


Synthesis of biological molecules on molecular sieves.

Catalytic properties of aluminosilicates may play a role in the synthesis of biological molecules from simple gaseous molecules commonly found in planetary atmospheres. Urea, amino acids and UV absorbing substances have been obtained by heating CO and NH3 with Linde molecular sieves saturated with Ca+2, NH4+ or Fe+3. The yields of amino acids produced have been determined by an amino acid analyzer. The quantity of urea produced largely depends on the nature of the saturating cation. Experiments using 14CO confirm that the amino acids are not due to contaminants adsorbed on the surface of the molecular sieves.


Ultrastructural aspects of the thyroid in a case of human congenital goitre with cretinism.

Transmission electron microscope and scanning electron microscope studies in a case of congenital goitre associated with cretinism revealed the existence of two cell types: the first with rounded endoplasmic cisternae and normal lysosomes; the second characterized by an abundance of lysosomes and/or lipofuscin granules. Intermediary aspects suggested that the second cell type was the progeny of the first type. As colloid droplets and apical pseudopods engulfing colloid were nearly absent, and as NBE127I was elevated in the plasma, it is suggested that iodoprotein secretion occurred by a process different from colloid phagocytosis as observed after acute TSH stimulation.


Colchicine-induced lesions in the rat duodenum.

In rats subcutaneous injection of colchicine resulted in sub-cellular necrosis within the rapidly proliferating cells in the duodenal mucosa. Similar lesions resulted from the administration of cytosine arabinoside hydrochloride and the effects of both drugs were ameliorated by treatment with cycloheximide. It is suggested that these lesions are a form of shrinkage necrosis involved in the deletion of growing stem-cells which, in the absence of mitotic division, results in mucosal atrophy.


Failure to induce autoimmune phenomena in mice by administration of highly purified C-type particles from NZB mice.

Highly purified C-type particles from milk of NZB mice were ineffective in the transfer of Coombs positive hemolytic anemia and cryoglobulinemia to neonatal BALB/cHeA and NZC/B1 mice. Injection of neonatal NZB mice with these particles did not change the frequency or time of onset of these phenomena. Antinuclear antibodies were found in all injected animals in the same frequencies as in the control groups.


Pulmonary pressure- volume relationships after corticotropin (ACTH) and salin injections in fetal rabbits.

ACTH mixed with barium sulfate injected into fetal rabbits on day 24 of a 31-day gestation was associated with increased lung maturity compared with lungs of noninjected controls when assessed by deflation pressure-volume curves and lung weight to body weight ratios measured on day 27. Fetuses injected with saline and barium sulfate also had accelerated (although somewhat less) lung maturation, perhaps from an ACTH-mediated response to stress. The ACTH group LA) maintained the largest lung volumes on deflation, followed by the saline group (B), the ACTH controls (C), and the saline controls (D). Volumes (percentage of total lung capacity) between the two injected groups and their controls (A versus C; B versus D) were significantly different at transthoracic pressures of 15, 10, 7, 4, 0, and -2 cm water (p less than 0.05). Volumes (percentage of total lung capacity) between the ACTH group and the saline group (A versus B) and between the two controls groups (C versus D) were not significantly different. Wet lung weight to body weight ratios were significantly different between the injected groups and their respective controls, between the two injected groups, and between the two control groups.


Elevated urinary testosterone and androstanediol in precocious adrenarche.

Using a newly devised radioligand method for the simultaneous determination of urinary testosterone (T) and androstanediol (Adiol) nine girls with precocious adrenarche were evaluated. In the base-line state average urinary T excretion (1.29 mug/24 hr) and Adiol excretion (1.33 mug/24 hr) were significantly elevated when compared with 15 age-matched controls (0.3 and 0.33 mug/ 24 hr, respectively, P less than 0.001 for both). Adrenocorticotropic hormone (ACTH) infusion performed in five patients with precocious adrenarche produced at least a 50 greater than increase in urinary T excretion in all and a similar increase in Adiol excretion in four of five patients. Dexamethasone administration in the same five patients produced a 25 greater than fall in urinary T excretion in all and a comparable fall in Adiol in four.


Glucose production in the newborn dog. II. Evaluation of autonomic and enzymatic control in the isolated perfused canine liver.

The effects of birth and morepinephrine on hepatic glucose production, glycogenolysis, and gluconeogenesis were examined in livers isolated from fetal dogs at term, littermates 3 hr after delivery, and newborn dogs 1-5 days old. Livers were perfused in pairs with medium containing (6-3H)glucose (6 mM) and (3-14C)lactate (10 mM +/- a pharmacologic amount of norepinephrine (10(-6)M). Changes in glucose production rates were correlated with changes in the enzymatic activities controlling gluconeogenesis. Net glucose production was less than 0.48 mumol/min-g liver both fetal and 3 hr liver but stablized above 1 mumol/min-g later during the first day. Initially, mobilization of the fetal hepatic glycogen accounted for glucose output. Subsequently, incorporation of lactate into glucose rose from negligible fetal rates to 0.19 mumol/min-g and accounted for 21% of net glucose production on day 3. Mazimal pyruvate carboxylase activity and mitochondrial CO2 fixation increased postnatally and correlated directly with net glucose production, glucose production from glycogen, and glucose production from lactate. Fetal liver did not respond to norepinephrine. Thereafter, norepinephrine increase hepatic glucose production by stimulating glycogen breakdown. Postnatal acceleration of glucose production and the response to norepinephrine occurred only after indiction of mitochondrial CO3 fixation. During day 1 the decline of hepatic glycogen in response to norepinephrine correlated with both CO2 fixation and lactate incorporation into glucose. Thus, initiation of gluconegenesis after birth may have been required for the postnatal acceration of hepatic glucose production and for the regulation of glycogenolysis by norpinephrine.


Abnormalities of carbamyl phosphate synthetase and ornithine transcarbamylase in liver of patients with Reye's syndrome.

Urea cycle function was evaluated in liver obtained from six patients with Reye's syndrome and from five control subjects. Reye's syndrome patients demonstrated normal activities for the extramitochondrial portion of the urea cycle, but showed marked abnormalities of the mitochondrial enzymes, i.e., carbamyl phosphate synthetase (CPS) and ornithine transcarbamylase (OTC) (Tables 2,3). CPS activity was reduced to less than 15% of control values in all four patients from whom tissues was obtained during the first 72 hr after the onset of encephalopathy. Two patents from whom tissue was not obtained until after 9 days of symptoms showed no reduction in CPS activity. The OTC activity was also reduced (3-67% of control values) in the four patients from whom tissue was obtained early in the illness. In addition, greater than 60% reduction in Vmax and Km for carbamyl phosphate was noted in all four patients in whom sample size permitted kinetic analysis, including both patients in whom CPS and OTC activity were not markedly reduced. The same kinetic abnormality as well as decreased CPS activity were experimentally produced in normal rate liver incubated in the presence of 1.0 mM 4-pentenoic acid, a short chain fatty acid and known hepatic mitochondrial toxin (Table 4).


Longitudinal sleep patterns during pubertal growth: four-year follow up.

There were little or no indications of differences in sleep outcomes between the sexes. Results indicate a disturbance of sleep on initial laboratory nights relative to later nights. The results reported here clearly document the persistence of these effects from year to year. For the most part, sleep characteristics during the 4 years immediately after onset of puberty appear to represent a typical phase in the gradual patterns of changes across all ages. Total sleep time decreased markedly from 560 min in age range 10-12 to 424 min in age range 20-29, with our puberty subjects as intermediate levels. Puberty subjects has an average of 2.5 awakenings/night in the first 2 years as compared with 1.2/night in the last 2 years. The number of sleep stage shifts during the night varied around a constant mean value of approximately 37/night throughout all ages. The number of rapid eye movement (REM) period during the night decreased sharply for individuals from childhood (6.9/night) through adolescence (4.0/night), remaining constant thereafter. Percentages of the various sleep stages were fairly constant for individuals from age range 10-12 through age 30-39. Our puberty subjects had percentage profiles in near perfect agreement with the normal ontogenetic process. Normative data suggest that slow wave sleep reaches a peak at some point during the teen years.


Solubility of various inert gases in rat skeletal muscle.

For the determination of solubility coefficients, isolated rat abdominal muscles were equilibrated at 37 degrees C with various inert gases saturated with water vapor. After rapid transfer into a closed chamber containing room air the amount of gas extracted from the sample by diffusion was measured by gas chromatography. Corrections for unextracted gas and for gas lost during the transfer of the sample were applied. The following mean values for the solubility coefficients, in mumol-1(-1)-torr(-1), were found: chloro-difluoro-methane (Freon 22), 56.0; acetylene, 55.5; nitrous oxide, 27.7; methane, 2.42; hydrogen, 1.13; helium, 0.608; sulfur hexafluoride, 0.559. The relationships between solubility in tissue, in water and in olive oil are discussed.


Diffusivity of various inert gases in rat skeletal muscle.

Krogh's diffusion constant (K) was determined for various inert gases in isolated rat abdominal muscle at 37 degrees C by measuring the amount of gas diffusing per unit time and partial pressure difference through a portion of the muscle of known surface area and thickness. The following mean values for K, in 10(-9) mmol-min-1-cm-1-torr-1, were obtained: C2H2, 42.2; N2O, 20.0; CHClF2, 18.8; H2, 1.67; He, 1.42; CH4, 1.27; SF6, 0.081. From Krogh's diffusion constant, the diffusion coefficient (D) was calculated using the solubility coefficient determined previously in the same preparation. The D values thus obtained were found to be about half the D values in water at 37 degrees C. Model calculations show that for gases with high lipid/water partition coefficient, D in tissues containing lipid is underestimated by this method. Graham's law (inverse proportionality between D and square root of molecular mass) was found to represent a useful approximation for these gases. A better correlation, however, was obtained between D and the molecular diameter.


The role of extracellular Ca2+ and cyclic nucleotides in the mechanism of enzyme secretion from the cat pancreas.

The role of Ca2+ in protein secretion from the isolated perfused cat's pancreas, the effect of the dibutyryl analogues of cAMP and cGMP, and the interrelation of Ca2+ and the nucleotides were studied. The following results were obtained: 1. Pancreatic enzyme secretion can be elicited by CaCl2 injections into the pancreatic arteries and is linearily related to the peak Ca2+ concentration in the effluent perfusate. Different background Ca2+ concentrations in the perfusate (3 mM or 0.125 mM) do not disturb this relation, indicating that no adaptation occurs. The effect of Ca2+ injections is of the same magnitude as that evoked by the hormones pancreozymin or acetylcholine. 2. Injections of Ca2+ potentiate the effects of dibutyryl adenosine 3',5'-cyclic monophosphate (dbcAMP), dibutyryl guanosine 3',5'-cyclic monophosphate (dbcGMP) or theophylline. 3. Infusion of low doses of pancreozymin increases the Ca2+ effect. The findings indicate that extracellular Ca2+ is involved in the mechanism of enzyme secretion and that Ca2+ and cyclic nucleotides have a synergistic action on the target.


S-adenosylmethionine: DNA-cytosine 5-methyltransferase from a Novikoff rat hepatoma cell line.

Partial purification of DNA methylase from Novikoff rat hepatoma cells is described. Contamination with other proteins persists although the enzyme preparation has a high specific activity and is purified 980-fold over homogenate activity. Evidence suggests, but does not prove, that there may be more than one species of DNA methylase in these cells. The enzyme has two broad pH optima at pH 7.0 and 7.5 and most readily methylates heterologous denatured DNAs although complex reaction kinetics indicate that native DNAs may eventually be methylated to an equal or greater level. The preparation of undermethylated DNA from Novikoff cells is also described. Undermethylated homologous DNA is an 85-fold greater acceptor of methyl groups than fully methylated Novikoff cell DNA. In contrast to other DNA substrates, the enzyme preparation methylates native undermethylated homologous DNA at a 3.5-fold greater than denatured undermethylated homologous DNA.


The synthesis of the internucleotide (phosphodiester) bond by a base-catalysed reaction.

Potassium tert-butoxide in hexamethyl phosphoramide and dimethyl formamide provides an excellent catalyst for the reaction of a 3'-hydroxyl or a 5'-hydroxyl group of a nucleoside with an appropriate nucleoside phosphorofluoridate to yield the dinucleoside phosphate. This paper describes the experiments leading to the development of this reaction together with the synthesis of thymidylyl-(5' leads to 3')-thymidine (dT-dT).


Sequences spanning the EcoRI substrate site.

Substrate recognition by the EcoRI restriction endonuclease was investigated by analysis of the nucleotide sequences at the sites of enzymatic cleavage in various DNA molecules. 5'-end labeling and homochromatographic fingerprinting led to the determination of a 17-base-pair sequence spanning the EcoRI site of simian virus 40 DNA and a 15-base-pair sequence overlapping the EcoRI site of Col El plasmid DNA. Three other DNAs were similarly tested, although extended sequences were not determined in these cases. The EcoRI site was shown to be symmetric double-stranded equivalent of -N-G-A-A-T-T-C-N-.


Mapping the transcription site of the SV40-specific late 16 S mRNA.

This paper describes the purification of polysomal RNA from SV40-lytically infected CV1 (monkey) cells and separation of the two distinct classes of SV40-specific mRNA sedimenting at 16 S and 19 S. These classes have been hybridized with the whole SV40 DNA genome as well as with the SV40 Hind fragments. The results have permitted the mapping of SV40-specific late 16 S mRNA from approximately 0.945 to 0.175 map units.


Effect of various concentrations of protein in chick diet upon the metabolic enzymes of glycine and serine.

The effect of feeding low protein diets with and without 2% glycine or 2% L-serine to chicks upon the enzymes concerned in the matabolism of glycine and serine has been determined. D-3-phosphoglycerate dehydrogenase (EC 1.1.1.s), hosphoserine phosphatase (EC 3.1.3.3), and serine hydroxymethyltransferase (EC 2.1.2.1) activities were significantly increased in livers from chicks fed 75% protein diets as compared to liver enzyme activities from chicks fed either 24% chick starter, 2% or 25% protein diets. Phosphoserine phosphatase activity was significantly higher in kidney tissue of chicks fed 75% protein diets. Livers from chicks fed 25% protein diets had a higher activity for D-3-phosphoglycerate dehydrogenase and phosphoserine phosphatase than did those fed chick starter or 2% protein diets. D-3-phosphoglycerate dehydrogenase and phosphoserine phosphatase activities were higher in livers from chicks fed 2% protein, 2% protein + 2% glycine, and 2% protein + 2% L-serine diets when compared to those from chicks fed low protein diets with supplemental methionine or cysteine. Serine dehydratase (EC 4.2.1.13), glycerate dehydrogenase (EC 1.1.1.29) and hydroxypyruvate-P: L-glutamate transaminase activities remained constant in livers from chicks fed all experimental diets. The uric acid concentration was significantly increased in plasma from chicks fed the high protein diets which suggests that D-3-phosphoglycerate dehydrogenase, phosphoserine phosphatase and serine hydroxymethyltransferase activities were increased because of the high requirement for glycine in uric acid formation. The 75% protein diet provided three times as much glycine as the 25% protein diet which may have met the increased need for glycine for uric acid formation.


Mercuric chloride effects on the activities of some hepatic enzymes in chicks.

Three groups of 6 male chicks each were fed a commercial diet and were given drinking water which contained either 0, 150 or 300 mug. of mercury/ml. as mercuric chloride from hatching to 3 weeks of age. The chicks were killed, the livers were removed and weighed, and the activities of selected enzymes were measured in the 800 X gav supernatant fractions of the liver homogenates. Liver weights were depressed from control values in chicks receiving 300 p.p.m. mercury but not in chicks receiving 150 p.p.m. Fatty acid synthetase specific activity was depressed by both levels of added mercury, but microsomal fatty acid elongation was depressed only by 300 p.p.m. of mercury. Both levels of added mercury stimulated acid phosphatase specific activity. The speecific activities of cytochrome c oxidase, glucose-6-phosphatase and 6-phosphogluconate dehydrogenase were unaffected by added mercury. The data support the hypothesis that mercury administration does not result in generalized hepatotoxicity.


Search for potential amoebicides. Part IV. Some hydrazones derivatives of 2-nitro-4-5-dimethoxybenzhydrazide and of 1-4-bis-[alpha(o-acetylhydrazino) phenethyl] piperazine.

By the condensation of 2-nitro-4-5-dimethoxybenzhydrazide and of 1,4-bis[alpha(O-acetylhydrazino) phenethyl] piperazine with various carbonyl compounds 42 hydrozones were synthesissed. Out of them 26 compounds were tested against the axenic-culture of E. histolytica at conc. of 125 mug/ml but no significant amoebicidal activity was observed.


Preparation of carcinoembryonic antigen (CEA) containing significantly increased amounts of galactose and galactosamine.

CEA was prepared by combined isoelectric precipitation, ultrafiltration and column chromatography under controlled conditions of pH. The resulting immunologically active materials were higher in carbohydrate (85-87%), N-acetyl galactosamine (10-11.5%) and galactose (28-32%) content than that previously reported. Differences in amino acid yield were also noted; the concentrations of aspartate, serine, glycine and alanine being higher and that of lysine, histidine, arginine, proline, valine, isoleucine, leucine and tyrosine were lower than that reported for CEA prepared by previous methods. The tumor tissues for CEA extraction were obtained from two Group O Rh positive deceased. Neither preparation showed Group A or B activity as measured by hemagglutination inhibition. It is suggested that the method of purification influences the carbohydrate and amino acid yields.


Interaction of ubisemiquinone with a paramagnetic component in heart tissue.

The origin of most of the electron paramagnetic resonances obtained at low temperature and low microwave power from heart tissue and subcellular fractions derived therefrom is now understood. A signal that emerges on partial reduction with characteristic lines at 3227 G (0.3227 tesla) and 3309 G (0.3309 tesla) (at 9.2 GHz) and disappears again on full reduction has remained unidentified. According to its behavior on oxidation-reduction, the substance giving rise to this signal has the properties of a two-electron acceptor. The signal is strongly dependent on temperature and can only be well resolved at less than 20 degrees K. It is readily elicited in submitochondrial particles by partial reduction, but has not been observed in submitochondrial particles from which ubiquinone has been removed by pentane extraction. When ubiquinone is reincorporated into extracted submitochondrial particles, the signal is again easily produced by partial reduction. Electron paramagnetic resonance spectra of partially reduced submitochondrial particles recorded at 34 GHz show lines centered about g approximately 2 with the same separation (approximately 82 G; approximately 0.0082 tesla) as do 9.2 GHz spectra, whereas no lines are detected with a separation of approximately 82 X 34/9.2 G (0.0082 X 34/9.2 tesla). We suggest, on the basis of these observations, that the unidentified signal arises from an interaction of ubisemiquinone and a second paramagnetic species. Three obvious choices exist concerning this second species: ubisemiquinone, flavin semiquinone, or an iorn-sulfur center. It is not possible without much additional information to decide between these possibilities. Since we have never observed the signal in the absence of the membrane-bound, high-potential type iron-sulfur protein, we have considered involvement of this species in the interaction. However, according to computer simulations of the observed electron paramagnetic resonance spectra, which yield best fits for semiquinone-semiquinone interaction, the possibility that ubi- or flavin semiquinone is the interaction partner appears more likely at this time. The interaction appears to be of the magnetic dipole-dipole type, but it is not certain whether there is also a contribution from spin exchange coupling. If it is assumed that the signal is due to magnetic dipole-dipole interaction, the distance of the partners is less than or equal to 7.7 A.


[Determination of the relative volume of the incorporations of a lipid nature by Candida guilleirmondii yeast cultured on hydrocarbons].

The paper describes a method for measuring a relative volume of lipid incorporations into yeast Candida guilliermondii cultivated on hydrocarbons. A relative volume of the incorporations (mean ratio of the volume of lipid incorporations to the cell volume expressed as percentage) as compared with the hydrocarbon content in the cells expressed as percentage of the yeast dry weight is given for each yeast sample. A direct correlation between the hydrocarbon content and relative volume of lipid incorporations has been demonstrated. The distribution of lipid incorporations in yeast populations with a different content of hydrocarbons has been studied statistically.


Location of histones on simian virus 40 DNA.

The physical location of histone molecules in a simian virus 40 DNA-histone complex isolated from purified virions was examined using site-specific restriction endonucleases. The complex contains four host histone species but lacks histone F1. Histones prevent complete cleavage of SV40 DNA by two restriction enzymes, HindIII and EcoRI. From the pattern of DNA fragments resulting from cleavage of the histone-DNA complex by the HindIII endonuclease, which makes six breaks on purified SV 40 DNA, we have concluded that histones are randomly arranged on SV40 DNA relative to restriction enzyme cleavage sites. The EcoRI endonuclease, which makes one break in SV40 NDA, was used to determine the degree of physical coverage of the SV 40 DNA molecule by histones. We observed that 80% of the EcoRI sites in the complex are accessible to the enzyme while 20% are "closed." This degree of coverage is consistent with the mass ratio of DNA:histone in the complex as revealed by the buoyant density of the formaldehyde-fixed complex. We conclude that the histones in the complex are located randomly on the SV 40 genome and cover approximatley 20% of the DNA. These results suggest that the histone species F2b, F2al, F2a2, and F3 are bound without regard to nucleotide sequence of SV 40 DNA.


[Regulation of amylolytic complex enzyme biosynthesis in Endomycopsis fibuligera strain 20-9].

The effect of glucose and cyclic 3',5'-adenosine monophosphate (cAMP) on the synthesis of glucoamylase and alpha-amylase by the yeast Endomycopsis fibuligera str. 20-9 was investigated. The glucoamylase synthesis was inhibited by every concentration (0.05--3.0%) of glucose. The synthesis of alpha-amylase was significantly reduced by high concentrations (2.0-3.0%) of carbohydrate only. An addition of cAMP (1 X 10(-5) M) reversed compeltely the inhibitory effect of 0.5-2.0% glucose on the alpha-amylase synthesis and partially restored the glucoamylase synthesis. It is concluded that catabolite repression may be involved in the regulation of alpha-amylase and glucoamylase synthesis by Endomycopsis fibuligera str. 20-9 and that this type of regulation shows varying effectiveness in the enzyme synthesis.


[Effect of the method of cell disintegration on the aspartate kinase activity of preparations from Bacillus polymyxa var. Ross].

The influence of methods of cell disintegration of Bac. polymyxa on aspartate kinase activity (EC 2.7.2.4) was examined. The disruption by means of the Hews press yielded a more active preparation as compared with ultrasonic disintegration. The supernatant treatment with streptomycin sulphate increased the preparation activity 2-fold. Dialysis of the fraction with a high activity of aspartate kinase inactivated the enzyme by 80--85%. The relationship between the aspartate kinase activity and the portein and ATP concentration in the reaction mixture was established.


Hormonal effects on structure and catalytic properties of fructose 1,6-bisphosphatase.

Gluconeogenic conditions, such as administration of triamcinolone or alloxan diabetes, cause the following changes in the molecular structure and properties of rabbit liver fructose 1,6-bisphosphatase (D-fructose-1,6-bisphosphate 1-phosphohydrolase, EC 3.1.3.11): (1) the appearance of traces (about 10%) of a lighter subunit; (2) loss of tryptophan from all of the subunits, including those that show no apparent change in molecular weight; (3) increase in requirement for the positive allosteric effector, histidine; (4) increase in amount of enzyme, but not its specific activity. These changes are identical to those induced by cold or fasting, and are related to increased activities of lysosomal proteases. The results suggest that lysosomes may act as mediators of gluconeogenic stimuli.


Inactivation of rabbit muscle phosphorylase phosphatase by cyclic AMP-dependent kinas.

Partially purified rabbit skeletal muscle phosphorylase phosphatase (EC 3.1.3.17; phosphoprotein phosphohydrolase) was inactivated when it was incubated with exogenous cyclic AMP-dependent protein kinase (EC 2.7.1.37; ATP:protein phosphotransferase), cyclic AMP, and ATP-Mg. Subsequent separation of the phosphatase by acrylamide gel electrophoresis or sucrose density centrifugation resulted in reactivation of the enzyme. The phosphatase decreased in molecular weight from approximately 70,000 to 52,000, and a phosphorylated inhibitor with molecular weight of 26,000 was found. Reactivation of phosphatase also occurred when it was incubated with MnCl2 or trypsin. The inhibitor was effective at less than 10(-8) M and was relatively heat stable. Its activity was destroyed by tryptic digestion and by dephosphorylation by a Mn-stimulated phosphatase. These observations support the possibility that phosphorylase phosphatase activity is controlled by cyclic AMP-dependent protein kinase and a Mn-stimulated phosphatase by a reaction involving phosphorylation and dephosphorylation of a protein phosphatase inhibitor.


Interchain disulfide bonds in procollagen are located in a large nontriple-helical COOH-terminal domain.

Tadpole collagenase (EC 3.4.24.3) cleaved chick cranial bone procollagen into two triple-stranded fragments, PCA and PCB. Only PCB, with an estimated molecular weight of about 60,000 for each component chain after reduction, was found to contain interchain disulfide bonds. The analogous cleavage of collagen is known to produce a large NH2-terminal fragment with a molecular weight of 70,000 for each chain and a small COOH-terminal fragment containing chains of about 25,000 molecular weight. Since PCB was too small to represent the product NH2-terminal to the site of collagenase cleavage, localization of interchain disulfide bonds to a COOH-terminal domain in procollagen was indicated. This assignment was conformed by Dintzis-type short-term labeling experiments. Procollagen obtained by acid extraction of bone lacked the COOH-terminal disulfide-bonded domain. The findings support a model for procollagen consisting of three proalpha chains each containing nonhelical NH2-terminal extensions of 20,000 molecular weight and COOH-terminal extensions of about 35,000 molecular weight, the latter linked by interchani disulfide bonds.


Effect of chemical inactivating agents on glucocorticoid receptor proteins in mouse and hamster cells.

The ffect of N-ethylmaleimide and iodoacetamide on the glucocorticoid receptor activity extracted from the cytosol of either mouse of hamster cells has been investigated. Treatment of mouse or hamster cytosol with N-ethylmaleimide or iodoacetamide rapidly inactivates the [3H]glucocorticoid hormone binding activity of either cytosol. Prebinding the glucocorticoid hormone, dexamethasone, to the cytosol receptor blocks the rapid inactivation of the receptor by N-ethylmaleimide. Treatment of the prebound hormone-receptor complex with iodoacetamide prevents the subsequent binding of the hormone-receptor complex to DNA without causing a dissociation of the complex. Although the conclusions may be limited by the lack of purity of the receptor, the results suggest that a sulfhydryl group is involved in the binding of glucocorticoid hormones to the receptor protein. In addition, the results suggest that iodoacetamide is inactivating a separate chemical site which is necessary for the binding of the hormone-receptor complex to DNA.


Cyclic AMP-mediated induction of ornithine decarboxylase of glioma and neuroblastoma cells.

The activity of ornithine decarboxylase (EC 4.1.1.17; L-ornithine carboxy-lyase) of C6-BU-1 glioma and N115 neuroblastoma cells increases significantly when confluent cultures are treated with compounds that increase cellular cAMP levels. These include norepinephrine or isoproterenol, and prostaglandin E1 or adenosine, which stimulate ornithine decarboxylase activity in C6-BU-1 glioma and N115 neuroblastoma cells, respectively. Ornithine decarboxylase activity is also elevated in confluent C6-BU-1 glioma cells treated with dibutyrylcAMP and theophylline, or after the glioma cells are fed with a serum-depleted medium in the presence of catecholamines and inhibitors of cyclic nucleotide phosphodiesterase. The activity of the enzyme increases 500- to 1000-fold, 2-6 hr after stationary-phase N115 neuroblastoma cells are fed with a serum-free medium, supplemented with phosphodiesterase inhibitors, adenosine, or prostaglandin E1. This stimulation is antagonized by carbamoyl choline and is blocked by actinomycin D or cycloheximide. These results suggest that the synthesis of ornithine decarboxylase of C6-BU-1 glioma and N115 neuroblastoma cells is controlled by cAMP.


Human synoviocytes: activation and desensitization by prostaglandins and 1-epinephrine.

The human synoviocyte increases its intracellular adenosine 3':5'-cyclic monophosphate (cAMP) concentration significantly after incubation with prostaglandin E1, prostaglandin E2, or l-epinephrine. The cells can be desensitized to these same hormones. Hormone-induced desensitization is receptor site specific and associated with a significant increase in intracellular 3':5'-cyclic AMP phosphodiesterase (EC 3.1.4. 17; 3':5'-cyclic AMP 5'-nucleotidohydrolase) activity, whereas cAMP-induced desensitization is not hormone specific.


Surface ruffles as markers for studies of cell transformation by Rous sarcoma virus.

Confluent chick embryo fibroblasts infected with the Ts68 mutant of Rous sarcoma virus were examined by scanning electron microscopy at the permissive (36 degrees) and nonpermissive (41 degrees) temperatures for transformation. Infected cells shifted from 41 degrees to 36 degrees undergo a change in shape from elongated to rounded. This process is preceded by the appearance of surface ruffles on the cell. These surface ruffles are not observed on cells maintained at 41 degrees, appear as early as 0.5 hr after a shift to 36 degrees, and are common on cells maintained at 36 degrees. By 3.5 hr after the shift from 41 degrees to 36 degrees, cultures appear fully transformed by the criteria of cell roundedness and the presence of surface ruffles. This surface alteration of cells is the earliest event of those so far reported during the transformation process and is not dependent upon protein synthesis and extracellular plasminogen during the period of temperature shift.


Chemotactic signals induce cell differentiation in Dictyostelium discoideum.

Experiments carried out with the aid of cellophane membranes demonstrate that the morphogenetic block of certain nonaggregating, "aggregateless," mutants may be overcome by diffusible factors excreted by aggregating wild-type cells. The same differentiation process into aggregation-competent cell is observed if mutant amoebae are subjected to external 3':5'-cAMP pulses imposed at 5 min intervals. Wild-type amoebae also respond to cAMP pulses, since the onset of differentiation is more precocious in pulsed than in unpulsed populations. These data suggest that chemotactic signals act as an inducer of cell differentiation.


Target cells in bone for parathormone and calcitonin are different: enrichment for each cell type by sequential digestion of mouse calvaria and selective adhesion to polymeric surfaces.

Six populations of bone cells (populations 1-6) were obtained by sequential digestion of mouse calvaria with collagenase and trypsin. After release from the tissue, each cell population was cultured for seven days. Parathormone, but not calcitonin, elicited an increase in intracellular cyclic AMP in the cells of populations 4, 5, and 6. In contrast, both hormones elicited increases in cyclic AMP in populations 2 and 3 but had no effect on population 1. When the cells of population 2 were exposed to a Falcontissue culture polystyrene surface for periods of time up to 5 min, many cells adhered. The nonadhering cell population contained a lesser proportion of cells responsive to calcitonin, whereas the adhering population contained a greater proportion responsive to this hormone. Conversely, when the cells of population 2 were exposed to an acid-treated nylon surface, the nonadhering cells contained a larger proportion of those responsive to calcitonin and a smaller proportion responsive to parathormone. When those cells that were enriched for calcitonin responsiveness were examined, we found an increased proportion that exhibited an asymmetric bipolar morphology. These differed from large amorphous, often binucleate, cells which predominated in those populations that responded exclusively to parathormone. These results establish that bone contains at least two types of target cells--one that responds to parathormone but not calcitonin, the other that responds predominantly to calcitonin.


Enhanced representation of HL-A antigens on human lymphocytes after mitogenesis induced by phytohemagglutinin or Epstein-Barr virus.

The amount of HL-A antigens present on the peripheral blood lymphocytes of a single human donor was increased about 11-fold after stimulation with phytohemagglutinin and 36-fold after transformation with Epstein-Barr virus. This increase applied to all four HL-A specificities of these cells. The response to phytohemagglutinin was dependent on dose and was first observed at 12 hr of incubation. Measurements of the amount of surface membranes by geometry, by radioiodinatable surface proteins, and by 5'-nucleotidase (5'-ribonucleotide phosphohydrolase, EC 3.1.3.5) assay all indicated that the enhanced representation of HL-A antigens after stimulation by phytohemagglutinin or transformation by Epstein-Barr virus must be due to an increase in the density of these antigens on the cell surface.


Thyroid hormone binding by a component of mitochondrial membrane.

The thyroid hormone, triiodothyronine, has been shown to be bound by the intranuclear chromatin protein associated with active DNA, where it is believed to stimulate transcription. Evidence exists that the thyroid hormones have direct action not only on nuclei, but also on mitochondria. Threfore, specific proteins that bind thyroid hormones in the mitochondria should be demonstrable. Mitochondria were isolated from homogenized rat livers by sedimentation through 0.25 M sucrose solution, followed by washing four times to free them  of microsomes. Strong binding of thyroid hormones was observed in mitochondrial fractions prepared from both the membranes and the matrix. After incubation in an ice bath with increasing amonts of triiodothyronine with added tracer [125I]triiodothyronine, the matrix infrequently contained specific saturable receptor sites, but usually exhibited strong "nonspecific" interaction...


C-type virus particles in placenta of normal healthy Sprague-Dawley rats.

C-type virus particles were found on electron-microscopic examination in placentas from two out of four young healthy Sprague-Dawley rats. One of these specimens contained virus particles budding from the plasma membranes of cells in the junctional zone of the placenta, i.e., the region where the fetal and maternal cell layers meet. In the other placenta, immature and mature C-type virus particles were found among cell debris also in the junctional region. This observation adds another species of animals to those recently reported, such as rhesus and baboon monkeys, as well as humans, in which C-type virus particles were found in the placenta. The presence of C-type viicant in view of the fact that a considerable number of these animals develop spontaneously a variety of malignant tumors, occasionally also leukemia and malignant lymphomas; however, none of these spontaneous tumors reveals the presence of virus particles on electron-microscopic examination. The nature of virus particles detected in rat placenta remains to be determined. As a working hypothesis, it is possible to assume that they may represent the passage of latent, presumably oncogenic, viruses transmitted "vertically" from parents to offspring. In the course of this passage some of them may be formed, emerging temporarily from their latency, before losing their identity and being again incorporated into the cell genetic components.


Identification of the 5' end of Rous sarcoma virus RNA.

The 5'-terminal triphosphate of the 35S RNA isolated from Rous sarcoma virus is blocked by 7-methylguanosine in 5' linkage with the penultimate nucleoside which is methylated in the 2'-O-ribose position, a type of endgroup found in all animal mRNAs investigated during the past year. The specific nuclease-resistant terminal fragment of RSV RNA has the structure 7mG5'ppp5'GmpCp-. This finding supports the belief that RNA of Rous sarcoma virus represents a (+) strand messenger which may be directly translated upon infection.


Reversible effects of chaotropic agents on the proton permeability of Escherichia coli membrane vesicles.

Extraction of E. coli ML 308-225 membrane vesicles with chaotropic agents causes the vesicles to become specifically permeable to protons. As a result, the vesicles no longer generate a membrane potential, interior negative, and they do not catalyze respiration-dependent lactose or proline transport. Treatment of the extracted vesicles with various carbodiimides decreases the permeability of the vesicle membrane to protons, causing them to regain their ability to generate a membrane potential. By this means, active transport is completely reactivated. Exposure of the vesicles to carbodiimides prior to extraction with chaotropic agents makes transport activity impervious to the effects of the chaotropes.


Receptor-mediated shifts in cGMP and cAMP levels in neuroblastoma cells.

3':5'-cGMP levels of neuroblastoma N1E-115 cells increase as much as 200-fold upon activation of muscarinic acetylcholine receptors, resulting in intracellular cGMP concentrations greater than 600 pmol/mg of protein. The cells also have receptors for adenosine which mediate an increase in 3':5'-cAMP levels. Unexpectedly, prostaglandin E1 was found to increase the concentrations of both cGMP and cAMP. Carbamylcholine, adenosine, and PGE1 were added to cells separately and in pairs to determine the effect of one compound on cell responses to another. Reciprocal inhibition, unilateral inhibition, additive, and nonadditive responses were observed with respect to cGMP and cAMP levels when different pairs of receptors were activated simultaneously.


Comparison between growth characteristics of an Epstein--Barr virus (EBV)-genome-negative lymphoma line and its EBV-converted subline in vitro.

The GC-BJAB cell line, which carries the Epstein--Barr virus (EBV), was derived from an EBV-genome-negative lymphoma line (BJAB) by EBV infection in vitro [G. B. Clements, G. Klein, and S. Povey (1975) Int. J. Cancer, in press]. Both lines grow at a similar rate at 37 degrees but they differ at other temperatures. BJAB grows well at 34 degrees, 37 degrees, and 39 degrees. GC-BJAB grows at 37 degrees and 39 degrees, but grows poorly at 34 degrees. At 37 degrees, GC-BJAB cultures can be maintained at the viable state for a long time after having reached saturation density (approximately 10(6) cells per ml). In contrast, BJAB cultures die very soon after having attained similar maximum density. Since the identity of the two cell lines has been critically established [Clements et al.; E. Svedmyr and M. Jondal (1975) Proc. Nat. Acad. Sci USA 72, 1622--1626; G. Klein, to be published; J. Zeuthen, personal communication] the remarkable differences in their growth properties must be attributed to the EBV genome.


Ultrastructural localization of insulin receptors on adipocytes.

The method for preparing a stable, biologically active, covalently linked ferritin--insulin complex has been modified to provide a 25-fold increase in yield compared to the original procedure while reducing the molar fatio of ferritin to insulin to 1:1 from 40:1. Ultrastructural studies of isolated adipocytes revealed specific binding of ferritin--insulin to the cell surface in irregular clusters associated with the glycocalyx coating. The number of ferritin--insulin molecules observed was consistent with the number of sulin molecules observed was consistent with the number of receptors calculated from 125I-labeled insulin binding studies. The ferritin--insulin was not observed in the cytoplasm of the cell but was found on the convave side of surface connected vesicles. These surface connected vesicles were part of an alveolar-like system of plasma membrane invaginations which project in various directions in the cytoplasm and by thin sectioning can appear as pinocytotic-like microvesicles. The morphological observations on ferritin--insulin binding were supported by the finding that 125I-labeled insulin binding was almost exclusively localized to highly purified plasma membranes isolated by fractionation of adipocytes after incubation with 125I-labeled insulin. These data supported the theory that insulin did not need to enter a cell to cause biological effects and was consistent with the negative cooperativity concept of insulin binding to cell receptors.


In vitro infection of lymphoid cells by thymotropic radiation leukemia virus grown in vitro.

Murine lymphoid cells were infected in vitro with purified leukemogenic radiation leukemia virus (RadLV) produced by virus-induced lymphoblast cell lines. Thymocytes were shown to be highly susceptible to infection by the virus, whereas murine or other fibroblasts were refractory to it. Murine bone marrow and spleen cells were shown to be much less sensitive to infection by this thymotropic RadLV. By comparison, a B-tropic RadLV isolate (RadLV), propagated on a mouse fibroblast cell line, was noninfectious for lymphoid cells but infected fibroblasts. A correlation was shown to exist between in vitro infection of thymocytes, as assayed by immunofluorescence, and in vivo leukemogenicity of the thymotropic RadLV. This constitutes a rapid in vitro test for in vivo leukemogenicity of a natural lymphatic leukemia virus.


Central role for magnesium in coordinate control of metabolism and growth in animal cells.

The rate of DNA synthesis in cultures of chicken embryo fibroblasts is reduced by deprivation of serum, high population density, and other "physiological" effectors, through a reduction in the number of cells in the S-period of the cell cycle. The same effect can be produced by drastically reducing the concentration of Mg++ added to the medium. This effect is erratic, however, and better control of [Mg++] can be achieved with phosphorylated compounds which preferentially bind Mg++. Both ATP and ADP, at concentrations in the medium less than or equal to [Mg++], stimulate DNA synthesis in cultures, and at greater concentrations inhibit DNA synthesis by affe-ting the proportion of cells in the S-period. Sodium pyrophosphate, which strongly complexes Mg++, causes little stimulation of DNA synthesis at low concentrations, but causes a striking decrease at concentrations exceeding [Mg++] of the medium. The inhibition can be fully reversed by adding an excess of Mg++, and the kinetics of increase in DNA synthesis resemble those which follow the restoration of serum to serum-deprived cultures. Limitation of [Mg++] by pyrophosphate also reduces the rates of RNA and protein synthesis, 2-deoxy-D-glucose uptake, and lactic acid production to an extent comparable to the reduction caused by the removal of serum from the medium. A model for the coordinate control of metabolism, differentiated function, and growth through the activity of divalent cations is described. The compartmentalization of Mg++ within the cell serves as the key element in this coordinate control by regulating those metabolic pathways in which the rate-limiting steps are transphosphorylation reactions.


Molecular weights of adrenocorticotropic hormone in extracts of anterior and intermediate-posterior lobes of mouse pituitary.

The molecular weight of the adrenocorticotropic hormone (ACTH) activity in extracts of the separated anterior and intermediate-posterior lobes of the mouse pituitary was determined by gel filtration in guanidine-HCl. Following dilution or removal of the guanidine-HCl, ACTH activity was quantitated by both radioimmunoassay and bioassay. Extracts of the intermediate-posterior lobe contain approximately a tenth as much ACTH activity as extracts of the anterior lobe. In extracts of both the anterior and the intermediate-posterior lobes, about half of the immunological ACTH activity is similar in size to porcine ACTH (molecular weight 4000--5500). Two higher molecular weight forms of ACTH account for the remainder of the ACTH activity. About 40% of the immunological ACTH activity in anterior lobe extracts has a molecular weight of 6500--9000. Extracts of both the anterior lobe and the intermediate-posterior lobe contain ACTH activity with a molecular weight of 20,000--30,000. While this 20,000--30,000 molecular weight ACTH accounts for only 5% of the immunological ACTH activity in the anterior lobe extracts, it accounts for half of the immunological ACTH activity in extracts of the intermediate-posterior lobe. Extracts of an ACTH-secreting mouse pituitary tumor cell line (AtT-20/D-16v) contain the same three molecular weight forms of ACTH. Each of the three molecular weight forms of ACTH has a characteristic ratio of biological ACTH activity to immunological ACTH activity, independent of the source of the material (anterior lobe, intermediate-posterior lobe, or mouse pituitary tumor cell line).


Periodate and concanavalin A induce blast transformation of rat lymphocytes by an indirect mechanism.

When rat lymph node cells, rendered incapable of division by treatment with mitomycin C, were then reacted with sodium metaperiodate (NaIO4), they stimulated the transformation of untreated, syngeneic lymph node cells in vitro. Unequivocal evidence that untreated lymph node cells responded in this situation was obtained by means of sex chromosome karyotype analysis. In experiments in which the ratio of responder to stimulator cells was varied between the limits of pure responder and pure stimulator cells at a constant cell density, [3H]thymidine incorporation increased linearly to a maximum at a ratio of 1:1, and then decreased linearly. These results suggest a predominantly bicellular reaction in which one periodate-treated cell stimulates only one responder cell. In experiments in which stimulator cells were reacted with periodate, but not treated with mitomycin C, and then mixed at a 1:1 ratio with untreated, syngeneic lymph node cells, karyotype analysis showed that both periodate-treated and untreated lymph node cells responded in significant numbers. Lymph node cells were cultured at various cell densities under three different conditions: one group was made up entirely of cells treated with periodate; a second group consisted of a 1:1 mixture of cells treated with periodate and untreated cells; the third group contained normal cells cultured in the presence of soluble concanavalin A. Expressed as [3H]thymidine incorporated per 10(6) cells, a progressive increase was found at the lower cell densities for all three groups, and identical slopes (about 2.5) were obtained when the results were plotted on a full logarithmic scale. Mitomycin C-treated lymph node cells incubated with concanavalin A and then thoroughly washed were found to be capable of transforming untreated, syngeneic lymph node cells. Maximal [3H]thymidine incorporation also occurred at a ratio of 1:1. These results show that, similarly to periodate stimulation, mitogenic stimulation by concanavalin A probably operates through an indirect mechanism, analogous to a mixed lymphocyte reaction. It is possible that other mitogenic agents also operate indirectly.


Plasma membrane alteration associated with malignant transformation in culture.

The intramembrane organization of the plasma membranes of nonmalignant cells in culture has been compared by freeze-fracturing with that of virally-transformed malignant cells. No dramatic differences are present in the distribution of intramembrane particles in the plasma membranes of these cells when the cells are examined without fixation or with mild fixation (glutaraldehyde treatment) prior to freezing. However, a redistribution of intramembrane particles into aggregates occurs in the membranes of nontransformed cells after treatment with glycerol. The aggregation of particles is extensive in normal chick embryo fibroblasts, and less extensive in mouse 3T3 cells. The glycerol-induced particle redistribution is not inhibited at 4 degrees, but it is inhibited by pretreatment with 2.5% glutaraldehyde. A significant number of the cells remain viable after the glycerol treatment, and the process is reversible. Particle aggregation does not appear to be related to either growth rate or cell density. Transformed Rous sarcoma virus/chick embryo fibroblasts and simian virus 40/3T3 cells have few particle aggregates after glycerol treatment. The plasma membranes of chick embryo fibroblasts transformed with a mutant of Rous sarcoma virus (TS-68) that is temperature sensitive for transformation, have few particle aggregates when grown at the permissive temperature (37 degrees). Extremely prominent particle aggregates are present in the plasma membranes of cells grown at the nonpermissive temperature (41 degrees). These observations indicate that there is an alteration in the plasma membrane associated with viral transformation which is related to a glycerol-sensitive mechanism that controls the distribution of intramembrane particles.


Enhancement of immunity against murine syngeneic tumors by a fraction extracted from non-pathogenic mycobacteria.

The data reported here demonstrate that a preparation extracted from nonpathogenic mycobacteria such as Mycobacterium smegmatis and hereafter referred to as interphase material protected mice against Ehrlich ascitic carcinoma, L-1210 leukemia, and another syngeneic lymphoid leukemia. Furthermore, mice treated by this preparation were much less susceptible to endotoxins than when stimulated by BCG (bacillus Calmette-Guerin) or M. smegmatis cells. Moreover, guinea pigs treated by interphase material administered in Freund's incomplete adjuvant showed an increased immune response, yet their sensitivity to tuberculin was much weaker than that of controls sensitized with Freund's complete adjuvant. Finally, resistance to Columbia SK virus infection could be demonstrated when interphase material was administered to mice prior to virus challenge.


Tumor regression at an untreated site during immunotherapy of an identical distant tumor.

The effects of two immunotherapy regimens on the development of an untreated, uniformly lethal transplantable line-10 hepatoma in strain 2 guinea pigs were monitored during treatment of an identical tumor 10 cm away. Line-10 cells were injected intradermally simultaneously at each of two sites. When one site was treated 6 and 16 days later with rabbit antibody against guinea pig fibrin fragment E, the complete regression of the treated tumor, a 25--30% depression in the development of the untreated tumor, and an increased survival time were observed. In another group of animals, when one site was treated 5 days after tumor challenge with syngeneic or xenogeneic "tumor-immune" RNA in a regimen including syngeneic nonsensitive lymphoid cells and tumor-specific antigen, all animals survived after complete and apparently specific regression of the tumors at both the treated and untreated sites. For the RNA regimen, we have shown that immunotherapy of an intradermally established line-10 tumor results in complete abrogation of both the treated and a distant untreated tumor; i.e., demonstrating a systemic effect.


Relationship between A-type and C-type particles in cells infected by Rous sarcoma virus.

Chicken cells infected with avian RNA tumor virus often contain small cytoplasmic A-type particles which commonly exist as clusters of 50--100 particles when viewed in thin sections. These particles were found more consistently in Rous sarcoma virus-infected than Rous-associated virus-infected cultures, but were generally present in only a small fraction of the total infected cells. The results of the survey of cells infected with various strains of leukosis-sarcoma viruses led to the hypothesis that the A particles develop in cells undergoing cytopathological degeneration. The hypothesis explains also the evanescent nature of the appearance of these particles in infected cells. The application of immunoelectron microscopic methods using monospecific antisera against viral internal proteins revealed that the A particles contain components immunologically related to the proteins of C-type virus.


Analysis of oncornavirus RNA subunits by electron microscopy.

Subunits of oncornavirus (avian myeloblastosis virus) RNA were isolated from purified 60--70S viral RNA by heat dissociation. Molecules sedimenting at 35 S, assumed to be the major component of the viral genome, were visualized in the electron microscope and their lengths were statistically analyzed. The results indicate a rather heterogeneous population of molecules with five distinct, reproducible size groups, an observation that excludes the assumption of random degradation of the genome. In addition, molecules of 28 and 18S RNA, always present in oncornavirus RNA preparations, were examined with the same methods. Some of these molecules possess secondary-structure regions similar to those characteristic for ribosomal RNA.


Cyproheptadine and beta cell function in the rat: insulin secretion from pancreas segments in vitro.

Pancreatic islet cell vacuolization, hyperglycemia, and glucose intolerance develop in rats after oral administration of cyproheptadine (CPH). In order to determine whether these effects were associated with abnormal insulin secretion, pancreas segments from CPH-treated and control rats were compared for their ability to secrete insulin in response to several stimuli. Oral administration of CPH (45 mg/kg/day) to rats for 1 or 8 days inhibited glucose-mediated insulin secretion from pancreas segments obtained 3 and 24 hr after the last dose of the drug. Insulin secretion had returned to normal by 48 hr after drug administration. Intraperitoneal administration of the drug was less effective than oral administration in inhibiting in vitro insulin secretion. Other stimuli for insulin secretion (tolbutamide, glucagon, L-leucine, and dibutyryl 3',5'cyclic AMP), like glucose, were incapable of releasing normal amounts of insulin from pancreas segments of CPH-treated rats. CPH and a metabolite, desmethyl-CPH, inhibited glucose-stimulated insulin secretion when added in vitro to pancreas segments from control rats. This suggests that the inhibition of insulin secretion in pancreas segments taken from animals treated with CPH could be due, at least in part, to the presence of drug and its metabolite in the tissue. A previously observed reduction in the pancreatic content of insulin in CPH-treated rats may also contribute to the abnormal insulin release in animals given the drug.


Scanning electron microscopy of glucocrticoid-treated hepatocytes and hepatoma cells in culture.

The morphological effects of exposure to hydrocortisone have been examined in two cell lines of liver origin by scanning electon microscopy. In one of these, an aneuploid line derived from a Morris hepatoma, the presence of hormone results not only in a suppression of cell proliferation, but in a marked flattening of the cells and loss of surface microvilli; in the other cell line, a diploid line derived from adult rat liver, the suppression of cell division is less marked, and the morphological effects of the hormone are far less striking. While the suppression of cell division in both of these cell lines is known to be rapidly reversible upon the removal of hormone, the presence of hormone causes the hepatoma cells to assume both monolayer growth characteristics and a morphology resembling those of cells derived from normal liver.


Analysis of the renin-angiotensin system during fasting in adult male rabbits.

Caloric deprivation for 3 days in adult male rabbits induced significant increases in daily urinary Na+ excretion, urinary volume and fluid intake as previously reported. These changes were accompanied by: (a) a significant reduction in plasma renin concentration; (b) an unchanged plasma renin activity; (c) a marked increase in the plasma angiotensinogen concentration; (d) a significant reduction in plasma angiotensin I; and (e) a significant increase in plasma angiotensin II. In a separate group of adult male rabbits, 3 days of caloric deprivation significantly increased the amount of converting enzyme in pulmonary parenchymal tissue. These findings correlate with the previously reported enhancement of mineralocortical hormone secretion and limiting effect of the latter on the natriuresis of caloric withdrawal. Since the increased mineralocortical hormone secretion does not prevent the natriuresis, the possibility that these striking changes in the components of the renin-angiotensin system during caloric deprivation may exert intrarenal effects is discussed.


Fluorescent virus precipitin test.

A fluorescent virus precipitin test (FVPT) for the serologic identification of small particulate antigens such as viruses has been described. The test has several advantageous characteristics: (a) It is probably as sensitive as any serologic test (i.e., aggregates with dimensions of 0.2 mum are detectable; therefore, complexes containing as few as three large viruses would give a positive test). (b) Cultivation of the virus is not required. (c) Since an indirect test can be used, only a single fluorescent conjugate is needed to permit the detection of a number of viruses. (d) The indirect test can be used to detect antiviral antibody. (e) The FVPT is rapid and reliable. (f) Its simplicity should enhance its general acceptance and application.


Adenosine, AMP, cyclic AMP, theophylline and the action and production of erythropoietin.

Erythropoiesis, as measured by the uptake of 59Fe into plethoric mice, is stimulated by adenosine, AMP, cyclic AMP, and dibutyryl cyclic AMP, but not by cytidine, its nucleotides or cyclic GMP. This stimulation is erythropoietin dependent, because it is prevented by anti-erythropoietin. Theophylline neither stimulates erythropoiesis nor potentiates the action of erythropoietin on bone marrow cells in plethoric mice. Theophylline does potentiate the production of erythropoietin in rats following a frief hypoxic exposure but does not cause a similar increase in mice.


Absence of idoxuridine and persistence of herpes simplex virus in brains of patients being treated for encephalitis.

Thirty-two specimens of brain from five patients with encephalitis suspected to be caused by herpes simplex virus, type 1 (HSV-1) were assayed for antiviral activity. Each patient received 60-80 mg/kg/day of idoxuridine (IDU) by intravenous infusion. The antiviral assay does not measure anti-HSV-1 antibodies. Biopsies of brain in every patient taken before IDU was used, and portions of several regions of the brain at autopsy were available during courses of treatment in four of the five patients. The last patient died 7 days after completing treatment. A significant concentration of IDU (833 mug/ml) was measured transiently in the cerebrospinal fluid of one patient. Meninges and brains showed inflammatory changes. Within the sensitivity of the test (larger than or equal to 6 mug/g) all specimens contained no IDU. As given, IDU does not achieve therapeutic concentrations in human brain. Further clinical use of IDU in therapy of herpes simplex virus encephalitis is not indicated.


Evidence against phosphorylation of a cyclic AMP-dependent protein kinase from bovine tracheobronchial smooth muscle.

The data presented in this report are evidence against the autophosphorylation of the cyclic AMP-dependent protein kinase isolated from bovine tracheobronchial smooth muscle. This suggests that there may be a fundamental difference in the regulation in vivo of the protein kinases from bovine heart and tracheobronchial smooth muscle.


In vivo electron spin resonance in rats.

In vivo ESR studies have been made on Wistar and Sprague-Dawley rats using travelling wave helices implanted in the left lobe of the liver. Tissue regenerates completely around the Teflon-enclosed turns of the coil and histological studies have shown that it is composed of liver cells and connective tissue. Electron spin resonance studies of this tissue in vitro reported by Commoner and Ternberg in 1961 have demonstrated that it exhibits the 'tissue radical' signal. The implanted coils have detected, in vivo, a three-line spectrum due to a strong concentration of 4-hydroxy- 2,2,6,6-tetramethylpiperidine-1-oxyl free radical ('tempol'), a nitroxide spin label, injected either intramuscularly, intraperitoneally or intravenously into the experimental animal immediately prior to the measurement. They have also detected, in vivo, a similar three-line spectrum due to 'spin-labelled chlorpromazine', a tranquillizing drug to which this nitroxide spin label has been attached, injected intramuscularly or intraperitoneally into the animal immediately prior o the measurement. Work is currently in progress to increase the in vivo sensitivity of the implanted helices by menas of improved coupling techniques and the use of a time-averaging computer.


Correlation between in vivo and in vitro metabolic measurements. Maximum capacity for urea synthesis.

Estimations of enzyme activity in vivo have been or can often only be done at unphysiological conditions. A main biochemical goal is to correlate in vivo and in vitro measurements. A possible approach to this problem is presented based on forcing metabolic activity in vivo to the maximum for a certain metabolic sequence. Since the urea synthesis system, including maximal rates of enzyme activities, is well known, we have compared in vitro maximum rates for the individual enzymes of urea synthesis with in vivo rates as judged by urea levels in blood of rats given large amounts of protein. The excellent agreement found between the calculated maximum activities from in vitro measurements to the time needed to metabolize a protein overload is presented and comments made on its significance and on the importance of maintaining protein intake at moderate levels, for the capacity of the urea system is limited. Since the intake of large quantities of protein increases the urea level in blood and in other tissues and since high urea levels are somewhat deleterious "per se" and particularly due to equilibrium with cyanate, ingestion of excessive amounts of protein is at best expensive and possibly hazardous.


NMR in cancer. VII. Sodium-23 magnetic resonance of normal and cancerous tissues.

Sodium-23 magnetic resonance was performed on four types of cancers and six types of normal tissues of rats and mice. The spin-lattice relaxation time of the tumors was generally longer than that of the normal tissues, with the most marked difference occurring between rat liver (T1 = 6.5 msec) and Novikoff hepatoma (T1 =23.7 msec). Estimation of tissue sodium from the signal intensity of the resonance indicated that all four types of tumors contained more sodium than any of the normal tissues.


Effects of ovariectomy and estradiol injections on food intake and body weight in rats with ventromedial hypothalamic lesions.

Female rats with lesions of the ventromedial hypothalamus (VMH) were ovariectomized during the static obese stage after body weight levels had stabilized. Following ovariectomy, rats with VMH lesions showed smaller increases in food intake and less body weight gain than non-lesioned controls ovariectomized at the same time. Subsequently, the effects of peripheral injections of estradiol benzoate (EB) on feeding and body weight were examined. Ovariectomized rats with VHM lesions were also less responsive to exogenous EB treatment; they lost significantly less weight in response to estrogen than controls. EB caused a somewhat smaller reduction in food intake by the VMH group but this difference was not significant. Considered together the available data on changes in responsiveness to endogenous and exogenous estrogen following VMH lesions suggest a role for VMH estrogen receptors in the regulation of body weight, but these estrogen receptors may not modulate weight by directly altering food intake as previously suggested.


Characterization of nuclear and chromatin poly A-containing RNA.

Poly A-containing RNA was isolated from Novikoff hepatoma cell chromatin on nitrocellulose membrane filters after the cells were labeled with 32P-orthophosphate for 6 hours. Under denaturing conditions, the poly A-containing chromatin RNA had a sedimentation coefficient of approximately 8-18S. The presence of poly A tracts in the chromatin RNA fractions indicate that poly A addition occurs almost immediately after transcription. Poly A-containing chromatin RNA and nuclear RNA were approximately 1/5 as active as cytoplasmic poly A-containing RNA in the wheat germ translational system. Since chromatin poly A-containing RNA is the precursor of cytoplasmic poly A-containing RNA, further processing of this RNA prior to its attachment to polysomes may account for its increase in translational capacity.


Stimulus-response coupling in neurohypophysial peptide target cells.

Recent data on the effects of neurohypophysial peptides at the cellular level are discussed with respect to the two basic processes involved in peptide hormone action--i.e., specific recognition of the information contained in the hormonal molecule and the transformation of this information into a stimulus leading to the final biological response. Four main aspects of this general problem are considered. A. Hormone-Receptor Interaction: Recent contributions in this field concern partial analysis of the three-dimensional conformation of oxytocin and vasopressin moleculal cells of the mammalian kidney. Conformational analysis of oxytocin and vasopressin molecules leads to the conclusion that, in solution, these peptides probably have a compact and highly stabilized three-dimensional configuration. Models have been proposed that provide a valuable clue to the interpretation of structure-activity relationships among natural hormones and many structural analogues. Binding studies with tritiated oxytocin and vasopressin have permitted determination of the kinetic parameters of hormone-receptor interaction in amphibian epithelial cells and mammalian kidney. B. Stimulus Generation: The nature of the primary stimulus generated by hormone-receptor interaction is still unknown. In the epithelial target cells of the amphibian skin and bladder and of the mammalian kidney, one of the first consequences of hormone-receptor interaction is the activation of membrane-bound adenylate cyclase. Analysis of the correlations between hormonal binding and adenylate cyclase activation suggests that activation is a function of receptor occupation rather than of the number of hormonal molecules interacting with the receptor per unit of time. On medullary adenylate cyclase of pig kidney, the relation between receptor occupancy and enzyme activation was found to be complex and nonlinear. The effects of several agents (calcium, nucleotides) on receptor occupancy and adenylate cyclase activation have been described. In mammalian uterus and other smooth muscle target cells, there is no evidence for direct involvement of cyclic AMP in the contractile response to oxytocin and other neurohypophysial peptides.


Electroencephalographic studies on the development of tolerance and cross tolerance to mescaline in the rat.

Recordings of the electroencephalogram (EEG) and the electromyogram (EMG) were collected continuously from rats equipped with permanent cortical and temporalis muscle electrodes. Automatic injections of mescaline were administered through indwelling i.p. cannulas at an initial dose of 30 mg/kg every 6 hrs for the first 2 days. This dose was then increased to 60 mg/kg 6 hr which was given for the duration of the study. The initial injections of the mescaline induced an immediate desynchronization of the EEG and behavioral arousal of the rat, which endured for 2-3 hrs. After this time, slow wave (SW) sleep and rapid eye movement (REM) sleep episodes reappeared, with the return of regular alternations of the sleep-wakefulness cycle. Upon continued administration of the drug, partial tolerance to the arousal effects of mescaline developed, which was reflected by a gradual reduction in the latencies to onset of SW sleep and REM sleep. Rats rendered tolerant to mescaline in this manner were found to be cross tolerant to lysergic acid diethylamide (LSD) and N,N-diethyl-tryptamine (DET). In contrast, cross tolerance did not occur to amphetamine, which exerts similar arousal and EEG desynchronizing effects. These results agree with physiological and behavioral studies of tolerance and cross tolerance among hallucinogens and support the usefulness of the EEG as a quantitative indicator of central nervous system function.


Effects of promethazine on nocturnal sleep in normal man.

The effects of a phenothiazine, on sleep in ten healthy volunteers were investigated in two double blind polygraphic studies. The first part consisted of a single dose study with promethazine 50, 100, and 200 mg, using pentobarbital 100 mg as a reference substance. In the second part, four subjects spent 20 consecutive nights with nine drug nights (promethazine 100 mg), followed by a placebo withdrawal period of six nights, in the sleep laboratory. Promethazine showed a dose related REM-depressing effect with a greater decrease, the higher the dose. The placebo value was 20.7% REM of total sleep time and gave 16.3, 13.5 and 11.4 respectively, for promethazine 50, 100 and 200 mg. Pentobarbital gave 16.2% REM. There was also an increase of stage II and with the highest dose an increase of stage III + IV. An increase of REM-latency together with a decrease of REM-periods was also seen, and while pentobarbital gave a decrease in REM-density, promethazine did not cause any changes in the phasic REM-component. A REM-rebound was seen in the first night of withdrawal with an increase of per cent REM from 19.9%-25.1%. The mean for the whole withdrawal period was 23.1%. Promethazine in the highest dose, 200 mg, gave drowsiness and hangover effects in 14 nights out of 20. The REM-depressing effect of promethazine together with its relatively weak REM-rebound effect may explain its value in the treatment of withdrawal symptoms following abuse of alcohol and barbiturates.


Fluorometric and myothermic studies of cardiac energetics.

Energy production during isometric and isotonic contractions of cardiac muscle has been studied using the isolated papillary muscle of the rabbit at room temperature. Muscles were mounted on thermopiles to record heat production associated with small numbers of contractions and then were mounted in an organ bath for fluorometric recording of the level of reduction of mitochondrial NAD. After each twitch there was a decrease in the level of fluorescence followed by an exponential return to the resting level, or further decreases after subsequent twitches. The fluorescence-time integral (integral delta Fl-dt), or area enclosed by a fluorometric transient during and after a series of twitches, was found to vary in magnitude in a manner similar to the magnitude of the total heat production when the mechanical conditions of contraction were altered by varying the load. The influence of the metabolic state of the muscle was also studied by using either glucose or sodium pyruvate as the exogenous substrates in the bathing medium. When pyruvate was substituted for glucose, maximum external work, maximum isometric force, and maximum mechanical efficiency were all increased. The kinetics of oxidative recovery metabolism, either measured fluorometrically or derived from the heat production, was accelerated when pyruvate replaced glucose.


Adenosine 3',5'-monophosphate, the myocardial cell membrane, and calcium.

Catecholamine-sensitive adenylate cyclase, cyclic nucleotide phosphodiesterase, adenosine 3', 5'-monophosphate (cyclic AMP)-dependent protein kinase, kinase substrate, and phosphoprotein phosphatase have variously been reported to be present in preparations of myocardial cellular membranes that function in the movement of Ca2+ in and out of the cell and in intracellular Ca2+ translocations, indicating that these membranees possess the equipment for the formation and destruction of cyclic Amp as well as for the initiation, effectuation, and termination of a possible membrane action of the nucleotide. It has also been observed that phosphorylation of seryl residues of protein in sarcolemma- and sarcotubule-rich myocardial subcellular fractions by cyclic AMP activated intrinsic and extrinsic protein kinases confers upon these membran structures an enhanced ability to bind or take up Ca2+ and that dibutyryl cyclic AMP, like adrenaline, produces in intact cardiac muscle simultaneous increases in contractile force and in the uptake of extracellular Ca2+. These findings are suggestive of a second messenger role of cyclic AMP in the beta-adrenoreceptor-mediated actions of catecholamines on myocardial contractile force and relaxation, in which Ca2+ would serve as a third messenger and be subject, respectively, to more effective removal from its binding sites on troponin. An alternative interpretation regards Ca2+ and cyclic AMP as interdependent twin second messengers in the catecholamine-induced inotropism. Since the physiological meaning of the reported effects of cyclic AMP on isolated myocardial membrane preparations is far from established an instances of a dissociation between the effects of catecholamines on myocardial contractile force and cyclic AMP levels have been observed, there is still room for hypotheses that relegate cyclic AMP to a nonobligatory, at most, supportive role in the action of the catecholamines on cardiac contraction.


The relationship between adenosine 3',5'-monophosphate levels and systolic transmembrane calcium flux.

The effects of inotropic agents on systolic transmembrane Ca++ flux were studied in isolated perfused guinea pig hearts depolarized with 22 mM K+. After depolarization, inotropic agents were added to the perfusion medium in an attempt to resotre excitability and contractions to the heart. Electrophysiological studies have shown that if excitability can be restored to the heart under these conditions, Ca++ carries the inward current of the action potential. Agents which putatively act by increasing intracellular levels of adenosine 3',5'-monophosphate (cyclic AMP) (isoproterenol, histamine, theophylline, papaverine, dibutyryl cyclic AMP) and Ca++ were found to restore excitability and contractions to K+-depolarized hearts. Threshold concentration for restoration by isoproterenol was 2 nM, and the strength of contractions developed by the restored heart were directly related to the dose of catecholamine used. Ca++ restoration was abolished by Ca++ antagonists (D 600, verapamil) but unaffected by beta blockade, whereas isoproterenol restoration was abolished by both. Ouabain, glucagon, and the divalent cation ionophore, A32187, failed to restore excitability to depolarized hearts. Correlative studies of tissue cyclic AMP levels were done. These data suggest that cyclic AMP can activate action potential-dependent Ca++ influx channels in myocardial cells.


Changes in the electroencephalogram and REM sleep time during morphine abstinence in pellet-implanted rats.

Rats were prepared with permanent electrodes for recording the electroencephalogram (EEG) and the electromyogram (EMG) and made morphine dependent by the subcutaneous implantation of morphine pellets. Abstinence was then precipitated by removal of the pellets 72 hours later. The evaluation of continuous EEG and EMG recordings revealed a maximal reduction in the amount of REM sleep and a decline in its EEG voltage output during the first day after pellet removal. Both the duration of REM sleep and its mean integrated EEG voltage returned to the baseline levels by the second day of abstinence. A significant but short-lived REM rebound subsequently followed and was accompanied by a trend toward elevation of the mean EEG voltage. These changes are reminiscent of similar findings in rats withdrawn from morphine administered intravenously (Khazan and Colasanti, 1971, 1972). The smaller magnitude of changes in rats treated with morphine pellets, however, may suggest a lower degree of drug dependence.


[Future of endocrinology].

With regards to the future of endocrinology, the author displays a spectrum formed by the following "hot" subjects: A) From a research point of view: 1) Membrane and cytosomal molecular receptors. 2) Prohormones, with regards to: a) paraneoplastic and tumoral syndromes; and b) "messenger" or transitory hormones. 3) Metabolic clearings: a) Normal; b) in endocrine pathology and c) in pathology of transformation and elimination: liver and kidney. B) From a practical-empirical point of view: 1) Tests of fetoplacental function. 2) Clinical perspectives of the use of hormonal factors and provoked endocrinological clinics (iatrogenic endocrinology). 3) Growth factors and somatostatine. 4) Thymus hormones. 5) Gastrointestinal hormones. 6) The renine-angiotensine-aldrosterone system, the problems of hypertension and preservation of the hydric compartments. Natriuretic hormones.


[Pneumological consultation practice in an anti-tuberculosis dispensary].

The authors present six years experience in connection with the diagnosis and the medical assistance of non-tuberculous broncho-pneumopathies in an anti-tuberculous dispensary. The data presented include 3600 cases with various diseases of which acute pneumopathies represent 11%, chronic bronchitis 14%, and broncho-pulmonary suppurations 9%. It is estimated that such an office of pneumological consultations within an anti-tuberculous dispensary is efficient and can, in the majority of the cases, make a correct diagnosis and provide corresponding dispensarization. This is conditioned by the existence of a nucleus for radiological, bacteriological and functional investigation.


[Circumstances concerning the diagnosis of tuberculosis in a pneumology department].

The analysis of a group of 67 patients hospitalized for various bronchopneumopathies, in whom evidencing of the tuberculosis bacilli led to the diagnosis of pulmonary tuberculosis allows to emphasize the increased efficiency of bacteriological detection by repeated bacilloscopy and successive cultures. They also stress the necessity of increased attention for pneumopathies that develop over the age of 50 years. The frequency of errors, predominantly stemming from the base network (territorial polyclinics), most of the confusion being made with suppurative broncho-pneumopathies, makes necessary the dispensarization of this category of patients in antituberculous units. The 56 new cases and the 11 cases in whom re-positivation was detected illustrate the possibilities of the bacteriological diagnosis and the contribution of a section of pneumology in the detection of pulmonary tuberculosis.


[Association of active pulmonary tuberculosis and pulmonary neoplasms].

Six cases are presented (out of a total of 11) with sputum positive pulmonary tuberculosis associated with bronchopulmonary carcinoma. The carcinoma developed during the evolution of 9 cases of active tuberculosis that had received a long discontinous treatment with tuberculostatic drugs. Only two of the 11 patients could be operated because the other 9 were hospitalized in the terminal stage of the disease, with invasive tumors. These had neither been suspected nor investigated for neoplasia. The use of all available means of investigation in the early stages of cancer can be of considerable help in the early detection of this disease and may allow for the application of surgical methods. Exploratory thoracotomy, together with extemporaneous histological investigation, are much more useful for the diagnosis than the expectative attitudes, which lead to theloss of the patient. Positive sputum should not eliminate the possibility of the association of tuberculosis with cancer.


[Renal complications after rifampicin].

Six observations are presented of patients receiving intermitent rifampicin treatment (900 mg twice weekly) that developed acute renal failure during treatment (4 cases) or when treatment was taken up again, after an interruption of 6 months (2 cases). Following anuria of 4--14 days and nitrogen retention between 248 and 521 mg, the evolution was favourable. Clinical, biological and immunological data (anti-rifampicin antibodies were detected in 1/8 and 1/16 in 3 cases) suggest the immune origin of this type of renal accidents.


[2 cases of hemolytic complications during intermittent treatment with rifampicin].

Two cases are presented, of hemolytic jaundice, determined by rifampicin. The first occured in a woman in whom treatment was started again after an interruption of 6 months and the second in a man undergoing intermittent treatment that had been tolerated very well for 3 months. Following the extension of the application of rifampicin in intermittent treatment might occur more frequently. Posology apparently has a major role and some precautions are to be taken.


[Adenitis following BCG vaccination].

The authors present a study on 167 cases of post-vaccinal adenitis after B.C.G. administration. These occured after the experimental application in 1,187 newborns of two vaccines produced by different laboratories. Manifest adenitis developed only after the application of the vaccine with a higher virulence and a significant concentration of live germs (A), while the vaccine with a low concentration of live germs only determined inflammatory adenitis (vaccine B). In the cases when vaccine A was applied, both after intra-dermal introduction of a 0,10 mg dose or of a 0,05 mg dose, it determined adenitis with a long evolution and in 7,79 percent, respectively in 6,95 percent of the vaccinated children fistulization occured. In most of the cases adenitis developed in the first 6 months following vaccination (86,77 percent) but in some cases it appeared even after 1-2 years. In 78,3 percent of the cases adenitis had a long course and in only 21,7 percent they regressed rapidly. Adenitis did not involve the general condition of the children but determined a concern of the parents. The duration of evolution of adenitis was, as a general rule, of several months, or 1-2 years and even longer and the treatment with tuberculostatics (INH) did not significantly alter their evolution. When there was a tendency to the formation of abscesses, especially in the case of gigantic adenitis, a reduction of the duration of the evolution was obtained by puncture, and especially by lymph-node curettage. The intensity of the post-vaccinal allergy is noted in cases complicated by adenitis (100 percent), as well as its long persistance, while in non-complicated cases the allergy diminished each year both in intensity and frequency. The authors recommend that before the mass use of a vaccine this should be tested, especially in young children, in view of making the correct choice of the efficient dose, with a maximal efficiency and the lowest number of complications.


[Comparative study of spirographic values and scintigraphic aspects in patients with bronchopulmonary diseases].

A correlation was made of routine spyrographic data with the scintigraphic aspects of the lungs in 146, respectively 100 patients with bronchopulmonary diseases. The following conclusions can be inferred: 1) normal spirographic values do not necessarily reflect an integrity of the respiratory function. Besides these values pulmonary perfusion can be severily altered; 2) the more deteriorated the spirographic values the more probable will be the alteration of the pulmonary perfusion; 3) in the evaluation of the respiratory function one cannot base on the routine spirographic data if these are within normal limits, especially before pulmonary surgery is performed, or before the patients are resuming active life; according to possibilities, the scintigraphic investigation is to be performed.


[Incidence of chemical diabetes in non-specific bronchopneumopathies].

In the present conditions of industrial development and of urbanization encountered in our country clinico-statistical observations show that the risk of disease can be doubled by the increased incidence of diabetes and of bronchopneumopathies with common germs. The much higher incidence of the metabolic disturbance in the asymptomatic, as well as in the clinically manifest stages of bronchopulmonary suppurative diseases, as compared with tuberculosis demonstrate the more severe evolution, as well as the medico-social implications that tend to overtake, from the viewpoint of the severity, those known in pulmonary tuberculosis, which shows a continuous decline. The early detection of both diseases, which may prevent their association, complications or reciprocal aggravation, represent an imperative task.


[Hyperchronic bronchopulmonary suppurations (clinical study of 100 cases)].

Following an analysis of the clinical observations files the authors present a series of aspects and clinical data, as well as functional results in connection with the evolution in out-bursts of 100 cases with bronchopulmonary suppurations dating for more than 10 years. These anatomo-clinical entities, developed either in healthy or on pre-existing lesions, of polymorphous aspect and frequently invalidating, with evident tendencies to relapses, determining considerable difficulties in the application of therapeutical measures, make necessary the dispensarization of patients and this must be organized in the frame of the network for pneumophthysiology, in view of providing the high quality assistance that is necessary in such cases.


[Model of eradication of tuberculosis in children in an area with high epidemiological potential].

In the territory of the Drajna TB dispensary, in the Prahova County, a series of complex measures have been experimented in the course of the second trimester of 1972, intending to prevent and to eliminate tuberculosis in children. Of the 31,731 children aged between 1 and 14 years, 29,496 have been tested with 2 units of PPD-IC65 (93 percent). A proportion of 12,6 percent of those tested presented reactions of more than 10 mm in diameter and 22,779 were vaccinated with BCG (Pasteur-Paris strain). The radiological pulmonary control of the reactors allowed for the detection of 24 cases of tuberculosis all of them new and asymptomatic. The efficiency of the detection was of 0,8 new cases per 1,000 tuberculin tested children or of 0,4 new cases per 100 of reactors that had been x-ray examined. This prophylactic action was repeated in 1973 in 2,219 children aged between 1 and 2-1/2 years that had been ommited in the preceding year and 4 new cases of tuberculosis were detected. In the medical districts in which there was a high prevalence of the children infection total radiophotographic case finding of the adult population was carried out. The result was a doubling of the TB incidence in children in 1972 followed by a 5-fold reduction of the indicators in 1973. The severe forms and death through tuberculosis in children have disappeared. The risk to develop the disease in BCG vaccinated children was reduced to 4,4 0/0000 per year. It is estimated that a generalization over the whole of the county of these prophylactic measures, as applied in the Drajna territory, under the form of a mass campaign, will allow for an improvement of the epidemiometric indicators in children in a relatively short time.


[Some epidemiological indicators in bronchopulmonary cancer in Sector 5 of Bucharest (study of 525 cases)].

The incidence was studied of broncho-pulmonary carcinoma, as well as of other epidemiological indicators in the 5th district of Bucharest over a period of 14 years (1960-1973) during which a total of 525 cases have occured. The incidence in this territory tends to become real from 1967 when the curve overtook the death-rate in the country. Following a continuous increase the number of cases reached 29 0/0000 in 1973 and it is presumed that it will attain 40 0/0000 in 1980. The increase in the morbidity is due to a higher number of cases noted in the extreme age groups (the increase was 2,5-fold in those under 40 years of age); the age group which was the most seriously involved was that between 60 and 69 years, in which the morbidity was of 111 0/0000. The proportion of women has also increased, from 1/6-th to 1/5-th of the total number of cases. In women the frequency of adenocarcinoma and of non-differentiated neoplasies is somewhat higher than in men. The proportion of histopathological confirmation of the disease diminished in the last years probably due to the higher number of aged subjects that cannot be correctly investigated or surgically treated.


[Radiophotographic and simultaneous bacteriological detection in an industrial group].

After carrying out a complex investigation of the workers of a confections factory the authors detected 2 new cases of tuberculosis (0,5 0/00), with the aid of radiophotography and another 2 cases (3,4 per 1000 subjects examined) by bacteriological investigation of the subjects that coughed. These last 2 cases were detected between those that had a normal thoracic aspect on the radiophotographic film alone. The observation of the authors allows to conclude that a bacteriological investigation of the suspects following radiophotography is not sufficient and that this type of investigation should be extended to certain groups of the population that show broncho-pulmonary symptoms. The association of the two methods, in well-defined technical and organisational conditions will allow for a significant increase of the efficiency of systematic examinations for the detection of pulmonary tuberculosis.