[Electron microscopic data on the mesosomelike and myelinlike structures of blue-green algae]. The mesosome-like structures of blue-green algae were studied by freeze-etching without preliminary fixation and, in parallel, by a technique of ultrathin sections. These structures were found mainly in Synechococcus elongatus and Synechocystis aquatilis, less often, in Anacystis nidulans and Anabaena variabilis, and were not detected in Plectonema boryanum. The following types of membrane structures were encountered: (1) formation consisting of three- and five-layered membranes and resembling the lamellar mesosomes of bacteria; (2) complex structures consisting of five-layered membranes and separated from the cytoplasm by an electron-dense substance not found in bacteria; (3) myelin-like structures; (4) formations intermediate between the mesosome-like and myelin-like structures. These structures were not found to be strictly confined to the growth phase of a culture. Possible functions of the mesosome-like and myelin-like structures are discussed ct. those of bacteria.


Epidemiology of human rotavirus Types 1 and 2 as studied by enzyme-linked immunosorbent assay. To determine the relative importance of two known serotypes of human rotavirus, we developed an enzyme-linked immunosorbent assay to differentiate serotype-specific rotavirus antigen and antibody. Using this technic, we studied the epidemiology of the two serotypes in acute gastroenteritis. Seventy-seven per cent of 414 rotavirus isolates were Type 2, and the remainder were Type 1. The serotype distribution was similar in specimens from children in Washington, D.C., and other parts of the world. Sero-epidemiologic studies revealed that most children living in the Washington, D.C., area acquired antibody to both types by the age of two years. An analysis of children who were reinfected indicated that sequential infections usually involved different serotypes and that illness caused by one serotype did not provide resistance to illness caused by the other serotype. These results suggest that, to be completely effective, a vaccine must provide resistance to both serotypes.


Subsensitivity of the rat striatal dopaminergic system after treatment with bromocriptine: effects on [3H]spiperone binding and dopamine-stimulated cyclic AMP formation. Repeated daily administration of the dopamine (DA) agonist bromocriptine (15 mg/kg; s.cut.) to rats led to a time dependent decrease in the in vitro binding of [3H]spiperone to striatal membranes. Kinetic analysis of [3H]spiperone binding after 2 and 7 days of bromocriptine treatment showed a 25-50% reduction in the total number of binding sites with no changein their affinity for spiperone. There was also a decreased accumulation of cyclic AMP (cAMP) in striatal slices in response to DA after bromocriptine treatment. The DA-sensitive adenylate cyclase in striatal homogenates, however, remained unchanged in bromocriptine treated rats. There was also no change in cyclic nucleotide phosphodiesterase activity in striatal tissue after bromocriptine treatment. Furthermore, incubation of striatal slices in the presence of the phosphodiesterase inhibitor isobutylmethylxanthine did not alter the decreased cAMP response to DA after 2 days of bromocriptine treatment. These results suggest that a decreased number of DA receptor sites may be responsible for the reduced cAMP response to DA in striatal slices after bromocriptine treatment.


Reduction of the equilibrium binding of cardiac glycosides and related compounds to Na+,K+-ATPase as a possible mechanism for the potassium-induced reversal of their toxicity. The influence of potassium ions on the equilibrium state of the binding of cardiac glycosides and their derivatives to partially purified dog heart and rat brain enzyme preparations was studied in vitro. The addition of potassium to the incubation mixture containing enzyme preparation, 3H-ouabain, Na+, Mg2+ and ATP, at the time when the binding reaction is close to equilibrium, caused an immediate reduction of the bound drug concentration; the concentration apparently shifting toward a lower equilibrium state. The degree of the potassium-induced reduction in bound drug concentration was dependent on the potassium concentration and on the chemical structure of the compound. The binding of aglycones, pentacetyl-gitoxin and cassaine was affected to a greater extent than that of the glycosides. These data suggest that one of the mechanisms by which potassium antagonizes the toxic actions of digitalis on the heart is to reduce the drug binding to cardiac Na+,K+-ATPase.


Effects of lithium and thallous ions on sodium pump activity in the guinea-pig heart and their relationship to the positive inotropic action. It has recently been demonstrated that both Tl+ and Li+ produce concentration- and time-dependent positive inotropic effects in guinea-pig atrial preparations although Tl+ inhibits and Li+ stimulates isolated Na+,K+-ATPase in vitro. In order to elucidate the mechanism of the positive inotropic actions of these cations, the effects of Tl+ and Li+ on sodium pump activity were studied. Active 86Rb uptake in guinea-pig ventricular slices, an estimate of sodium pump activity, was highly sensitive to the inhibitory effect of the cardiac glycosides. Preincubation of slices with Tl+ caused a dose- and time-dependent inhibition of active 86Rb uptake. Similar concentration- and time-dependent inhibition of active 86Rb uptake was observed when Na+ in a Krebs-Henseleit solution was partially replaced with Li+. Lithium, however, stimulated a partially purified Na+,K+-ATPase in vitro. During heart slice incubation, Tl+ and Li+ accumulated in a time-dependent manner. This accumulation was not readily reversible when slices were transferred into Tl+- or Li+-free solutions. It appears that the inhibition of sodium pump activity is related to the positive inotropic action of these cations.


The opiate-like action of tilidine is mediated by metabolites. The analgesic effect of tilidine in rats is completely antagonized by the narcotic antagonist naloxone. Radioreceptor assays revealed, however, that the main metabolites of tilidine, nortilidine and bisnortilidine, rather than tilidine exhibit affinity to opiate receptors. These findings were confirmed in studies using the electrically stimulated guinea pig ileum and the mouse vas deferens. Chronic tilidine administration to rats caused a considerable degree of physical dependence, which was expected from the ability of the intact animal to metabolize tilidine. In the isolated ileum from chronically morphinized guinea pigs, both nortilidine and bisnortilidine fully substituted for morphine in preventing induction of withdrawal, indicating dependence liability of these metabolites.


[Interaction of heterosensory afferent impulses in the neurons of the thalamic associative nuclei]. The interaction of visual, auditory and cutaneous afferent signals on neurons of pulvinar (Pulv), posterior lateral (LP) and mediodorsal (MD) thalamic nuclei was studied in acute experiments on cats anaesthetized with a mixture of nembutal and chloralorse. The first (more frequent) type of the interaction was characterized by inhibition or inhibition followed by facilitation of the response; the second--by facilitation or facilitation followed by inhibition of neuronal activity. The capacity for interaction in MD neurons was less expressed than in cells of the Pulv-LP complex (P less than 0.05). The interaction could be better seen in three-sensory elements. Certain peculiarities were found in the interaction of afferent signals coming from different peripheral receptive areas in neurons of both structures.


[Reactions of cat hindbrain "locomotor strip" neurons to microstimulation]. Synaptic responses of single neurons in the locomotor strip" were recorded extracellularly. Neurons of the rostral part of the strip produced short-latency responses to stimulation of the mesencephalic "locomotor region". Neurons of the caudal part of the strip responded to microstimulation of other sites of the strip, rostral ones included. When the distance between the site of stimulation and a neuron along the strip was less than 2--3 mm, short-latency (1.2--1.6 ms) responses were recorded. Thresholds and latencies grew with the distance. Polysynaptic responses with a 3--4 ms latency could be potentiated when repetitive (30--40 pulses per s) stimulation was used instead of a single stimulus. The results suggest that axons in the "locomotor strip" are oriented in a rostrocaudal direction and give off collaterals to neighbour neurons. The "locomotor strip" can be an integrative centre "intercalated" between the rostral centers of the brain and the spinal cord.


[Hypogenic cholinergic neuronal route in the central nervous system]. Because of the extensive overlapping within the brain of arousing and hypnogenic neurons the method of electrical stimulation has proved unsuitable for mapping out specific hypnogenic structures. On the other hand, the method of localized chemical stimulation achieved by introducing minute crystals of chemical substances permits the dissection of adjacent neurons with different functions. The aim of this study was to map out the central hypnogenic pathways introducing minute crystals of acetylcholine, eserine or atropine in different brain regions through cannulae permanently implanted in the brains of cats. The behavioral effects produced by the chemical stimulus were monitored simultaneously with electrical recordings from the neocortex, olfactory bulb, enthorinal cortex, and eyes movements. It is concluded that there is a sleep system made up of a multisynaptic chain of short axon neurons with cholinergic synaptic transmission and which extends throughout all the levels of the neuroaxis.


Fatty acid composition of myelin lipids (cerebrosides, sulphatides and sphingomyelin) from normal human sural nerve, and changes in peripheral neuropathy. Fatty acids of cerebrosides, sulphatides and sphingomyelin from normal human sural nerve, by comparison with brain, show a diminution of long-chained fatty acids as well as unsaturated fatty acids. The findings in brachial plexus are intermediate. In cases of peripheral neuropathy from various causes sphingomyelin fatty acid composition reveals pronounced loss of long-chained fatty acids, a phenomenon that is probably unspecific and may be associated with all forms of demyelination. Problems associated with the limited amount of nervous tissue available from biopsy specimens are discussed.


A quantitative histological study of changes in neurons and glia in the Gunn rat brain. The rostral part of the anterior limb of the anterior commissure (RAL) and the indusium griseum of the brains of Gunn rats and Wistar rats were examined quantitatively to assess the effects of hyperbilirubinaemia on the developing nervous system. The number of glia in the RAL was significantly less in Gunn rats than in the Wistar rats. The number of glia in the indusium griseum was similar in both groups but the Gunn rats showed a decrease of about 20% in the number of neutrons in this region. Myelination appeared to be unaffected and apart from an apparent increase in the number of dense bodies in astrocytes and oligodendrocytes there were no obvious morphological changes in either glia or neurons.


Anterior pituitary function before and after trans-sphenoidal microsurgical resection of pituitary tumors. Anterior pituitary lobe function was measured pre- and postoperatively in 40 patients with pituitary tumors who were managed surgically by the trans-sphenoidal approach. Of 23 patients with normal anterior lobe function preoperatively, 21 (91%) were normal postoperatively. Of 17 preoperatively impaired patients, 7 (40%) regained normal function, 3 (18%) improved, 4 (24%) remained the same, and 3 (18%) were further impaired by the surgery. In this small series, the likelihood of recovery of pituitary function varied inversely with the degree of preoperative impairment and the size of the tumor, suggesting that more aggressive surgical management of small, minimally symptomatic pituitary tumors is justified.


Adenoid cystic carcinoma of the Cervix. Report of 9 cases and a reappraisal. The clinical and pathologic findings of 9 patients with adenoid cystic carcinoma of the cervix are presented. A review of the literature identified an additional 29 cases. The tumor was usually associated with adenocarcinoma or squamous cell carcinoma of the cervix. Adenoid cystic carcinoma of the cervix appears to be an aggressive type of tumor; of the 38 cases reported in the literature lung metastases have occurred in 18 (47.4%), and of these, 14 patients are known to have died of the cancer.


Dual effect of external calcium on the frequency of miniature synaptic potentials in frog sympathetic ganglion cells. The effect of calcium on spontaneous transmitter release and on the release induced by tetanic stimulation and by raising the external potassium concentration ([K]0) was studied in sympathetic ganglion cells of Rana esculenta. 1. In standard Ringer's solution the frequency of miniature excitatory postsynaptic potentials (mepsp) ranged from 0.05--2.0 s-1 (0.05 +/- 0.09 s-1, n = 37) at room temperature. 2. At a [K]0 of 2.5 mM mepsp frequency was approximately linearly related to the logarithm of the external calcium concentration (log [Ca]0) (0.1 mM less than or equal to [Ca]0 less than or equal to 20 mM). 3. Duration and amplitude of the potentiation of transmitter release after tetanic preganglionic stimulation increased depending on [Ca]0. 4. Mepsp frequency was strongly dependent on [K]0 between 10 and 20 mM; the frequency being increased to about 40 times control level at a [K]0 of 20 mM. 5. Raising [Ca]0 up to 1.8 mM in high K solutions resulted in an increase in mepsp frequency followed by a decrease at higher [Ca]0. 6. These results are consistent with the idea that the effect of calcium on mepsp frequency depends on: (a) the driving force for calcium entry, (b) the effect of Ca ions on the potential gradient within the nerve membrane.


NAD in muscle of man at rest and during exercise. NAD can be used to assess the adequacy of oxygen availability to the respiratory chain. An enzymatic assay was established for NAD in human muscle biopsy samples. It gave reliable, reproducible results. The variation within and between subjects was less than 12%. Muscle NAD and lactate were determined at rest, and after bicycle ergometry work requiring approximately 75 and approximately 100% VO2 max (six subjects, four tests each). A positive (P less than 0.01) linear relationship between resting muscle NAD and percent slow twitch fibers was found, suggesting that fiber types may have different NAD content. Muscle NAD decreased during submaximal and maximal work (P less than 0.05). A large portion (73%) of the NAD reduction could be accounted for by increased muscle water. No relationship could be established between NAD and lactate. The negative linear relationship (P less than 0.01) between the muscle/blood ratio and percent slow twitch fibers is another indication of the fiber having different metabolic responses to the activity.


[Sterologic evaluation on in vitro thrombin-induced platelet degranulation and contraction (author's transl)]. Stereologic study of the in vitro release reaction from unstirred washed platelets provided quantitative information on the contractile wave induced by thrombin. Contraction was estimated from the reduction in the volume of both the granular compartment and the vacuoles of the surface connected canalicular system. Dibutyryl cyclic-AMP (10(-4) M) and dipyridamole (10(-3) M) inhibited degranulation, but they had no effect on the contraction produced by thrombin at high concentrations (0.1 UI/ml). Aspirin (10(-4) M) inhibited contraction induced by thrombin at concentration of 0.1 and 0.02 UI/ml (p less than 0.01 at each concentration). In two patients with hereditary cyclo-oxygenase deficiency, contraction produced by thrombin at low concentration (0.02 UI/ml) failed to induce contraction.


Characterization of two SV40 early mRNAs and evidence for a nuclear "prespliced" RNA species. Using in vitro translation of sucrose-gradient fractionated cytoplasmic mRNA from SV40-infected cells, we have shown that a deletion in the region mapping between 0.54--0.59 reduced the size of mRNA for small-t but not the size of mRNA for large-T. Mutants with a deletion in this region were shown to produce in vivo either shortened small-t or no small-t, and normal large-T. Similarly, in vitro translation of poly(A)+cytoplasmic RNA from cells infected with these mutants gave the same results. On the other hand in vitro translation of poly(A)+nuclear RNA from the mutants which made no small-t produced a small-t derivative possibly synthesized from a prespliced RNA species. We have also shown that poly(A)+nuclear RNA from mutant dl 2122 produced two small-t related proteins: one of these (MW: 11K) probably represents the product of a "prespliced" RNA, the other (MW: 17K) which is also found in the cytoplasm represents the product of the mutant specific small-t mRNA.


DNAse footprinting: a simple method for the detection of protein-DNA binding specificity. A method for studying the sequence-specific binding of proteins to DBA is described. The technique is a simple conjoining of the Maxam-Gilbert DNA-sequencing method and the technique of DNAase-protected fragment isolation. Fragments of a 5' end-labelled, double-stranded DNA segment, partially degraded by DNAase in the presence and absence of the binding protein, are visualized by electrophoresis and autoradiography alongside the base-specific reaction products of the Maxam-Gilbert sequencing method. It is then possible to see the protective "footprint" of the binding protein on the DNA sequence. The binding of lac repressor to lac operator is visualized by "footprinting" as an example. Equillibrium estimates indicate that 10-fold sequence-specificity (differential binding constant) could be studied easily using this technique.


Requirement for 7-methylguanosine in translation of globin mRNA in vivo. The 7-methylguanosine (m7G) residue present in the m7G5' ppp5'X-"CAP" structure of rabbit globin mRNA was removed quantitatively by periodate oxidation followed by beta-elimination in the presence of cyclohexylamine. The RNA thus treated was intact and exhibited no signs of degradation as examined by polyacrylamide gel electrophoresis in formamide. Assay for protein synthesis using a wheat germ cell-free system showed that the globin mRNA lacking m7G had lost most of its messenger activity. Identical treatment, of satellite tobacco necrosis virus (STNV) RNA, which does not contain the 5'-terminal "CAP" structure, resulted in no loss of its mRNA activity. Since the importance of the m7G residue in eukaryotic mRNA has not yet been shown essential for translation in vivo, both untreated and treated globin mRNAs were injected into frog oocytes and their translation into globin was measured at intervals over a ninety-six hour period. Globin mRNA either treated with periodate alone or lacking in m7g altogether were both found to have lost more than 90% of their activity in vivo.


Sexually transmitted diseases. Advances in management. We have briefly presented practical approaches to the management of the most common disorders caused by sexually transmitted pathogens, including pelvic inflammatory disease, urethritis in men, genital lesions, vaginal discharge, and syphilis. Guidelines for the management of gonorrhea may change if penicillin-resistant strains become more prevalent. Areas which require further research include the treatment of genital herpes and the prevention of recurrences, the treatment of nonspecific vaginitis, and definition of the importance of metronidazole toxicity.


Special investigations in COCM: biochemical analysis of cardiac biopsies. The introduction of the endomyocardial biopsy method has permitted unique possibilities to investigate the myocardium. Compared to several other methods described, it is superior for taking serial biopsies on one or more occasions. With already available methods, examination of heart muscle biopsies can be extended to a wide variety of biochemical analyses. The ultimate application of repeated or periodic tissue sampling is the potential of combining static and dynamic studies.


Coordination environment and fluoride binding of type 2 copper in the blue copper oxidase ceruloplasmin. The electron paramagnetic resonance (EPR) spectra of the blue copper oxidase ceruloplasmin [ferroxidase, iron (II):oxygen oxidoreductase, EC 1.16.3.1] and of a derivative having the type I (blue) copper centers reversibly bleached are reported. The EPR spectrum of bleached ceruloplasmin has a seven-line superhyperfine structure in the g : formula: (see text) region that is attributed to the presence of three nitrogen-donor type 2 copper ligands. The EPR data suggest further that the type 2 copper in ceruloplasmin possesses a tetragonal coordination gometry. In the presence of varying amounts of fluoride, superhyperfine splitting patterns in the g : formula: (see text) region of both ceruloplasmin derivatives indicate that a maximum of two fluorides may be bound to the type 2 copper.


[Obtaining iodinated luteinizing hormone while preserving its biological properties]. The author describes a method of obtaining biologically-active 125I-labeled luteinzing hormone in iodation with chloramine T. The hormone and chloramine T concentration ratio of 1:2, and the reaction time of 20 sec was used. LH-125I with the specific activity of 27--30 microCi/microgram was bound with the receptor strictly specifically.


DNA modifying enzymes of Agrobacterium tumefaciens: effect of DNA topoisomerase, restriction endonuclease, and unique DNA endonuclease on plasmid and plant DNA. Extracts from Agrobacterium tumefaciens strain ID135 contain three enzymes that have been characterized and partially purified. The first enzyme, a DNA topoisomerase, appeared to relax only negatively twisted DNA. The second enzyme, Atu I, a type II restriction endonuclease, generated the identical DNA digestion pattern as EcoRII when several DNAs were used. The third enzyme, endonuclease A, showed a preference for superhelical DNAs as substrates. When plasmid pCK135DNA, obtained from the virulent strain IDI135 of A. tumefaciens, or plant DNA was exposed to the three enzymes, changes in DNA patterns were observed due to either conformational changes or digestion of the DNAs. These enzymes may function in vivo in the processing and incorporation of bacterial DNA in plant cells.


Nucleotide sequences related to the transforming gene of avian sarcoma virus are present in DNA of uninfected vertebrates. We have detected nucleotide sequences related to the transforming gene of avian sarcoma vius (ASV) in the DNA of uninfected vertebrates. Purified radioactive DNA (cDNAsarc) complementary to most of all of the gene (src) required for transformation of fibroblasts by ASV was annealed with DNA from a variety of normal species. Under conditions that facilitate pairing of partially matched nucleotide sequences (1.5 M NaCl, 59 degrees), cDNAsarc formed duplexes with chicken, human, calf, mouse, and salmon DNA but not with DNA from sea urchin, Drosophila, or Escherichia coli. The kinetics of duplex formation indicated that cDNAsarc was reacting with nucleotide sequences present in a single copy or at most a few copies per cell. In contrast to the preceding findings, nucleotide sequences complementary to the remainder of the ASV genome were observed only in chicken DNA. Thermal denaturation studies of the duplexes formed with cDNAsarc indicated a high degree of conservation of the nucleotide sequences related to src in vertebrate DNAs; the reductions in melting temperature suggested about 3--4% mismatching of cDNAsarc with chicken DNA and 8--10% mismatching of cDNAsarc with the other vertebrate DNAs.


Antagonists of DNA gyrase inhibit repair and recombination of UV-irradiated phage lambda. Intracellular lambda DNA (from EDTA-sensitive tandem duplication phages) was extracted from infected rec+ bacteria and scored for infectivity and recombination (loss of duplication) by transfection of recA recB spheroplasts and subsequent assay for EDTA resistance. When phage development was blocked by repressor or by antibiotics (chloramphenicol and/or rifampin), the apparent recombination frequency was about 0.1% above the background value for recA infections. Prior irradiation of the phage greatly stimulated recombination; the frequency was 20% when UV fluence was 140 J/m2. Repair (recovery of infectivity) and recombination of irradiated phage DNA proceeded readily in the presence of chloramphenicol and rifampin. Inhibitors of DNA gyrase (coumermycin and oxolinic acid) blocked repair and reduced recombination. UV-stimulated recombination was very low in recA but nearly normal in recB cells: repair was reduced in both mutant strains. The recombination remained high as phage/cell ratios less than unity.


Alterations of hepatic Na+,K+-atpase and bile flow by estrogen: effects on liver surface membrane lipid structure and function. Administration of the synthetic estrogen ethinyl estradiol (17alpha-ethinyl-1,3,5-estratriene-3,17beta-diol) decreases hepatic Na(+),K(+)-ATPase (ATP phosphohydrolase; EC 3.6.1.3) activity and bile flow to 50% and alters the composition and structure of surface membrane lipid in rats. Although the content of phospholipids was not changed by treatment, free cholesterol (130%) and cholesterol esters (400%) were increased in liver surface membrane fractions. These observations correlate with changes in membrane viscosity, as shown by electron spin resonance probes. Both rotational correlation time, using the isotropic probe methyl (12-nitroxyl)stearate, and the order parameter, determined by the anisotropic probe 5-nitroxylstearic acid, were significantly increased in liver surface membrane fractions from rats treated with ethinyl estradiol. Administration of Triton WR-1339, a nonionic detergent that corrects hepatic and serum lipid changes caused by ethinyl estradiol treatment, restored toward normal elevated membrane lipids and viscosity as well as Na(+),K(+)-ATPase activity and bile flow. Although restoration of normal liver surface membrane structure and function may be due to reversal of abnormal lipid composition, detergents also may directly alter membrane enzyme activity. Addition of Triton WR-1339 in vitro increased Na(+),K(+)-ATPase activity and reduced membrane viscosity of surface membranes from rats treated with ethinyl estradiol. Triton had no effect on either parameter in normal membrane preparations. Studies of membrane structure and function both in vivo and in vitro suggest that alterations in lipid composition may alter Na(+),K(+)-ATPase function and bile flow.


6-beta-bromopenicillanic acid, a potent beta-lactamase inhibitor. 6-beta-Bromopenicillanic acid, which arises from the epimerization of 6-alpha-bromopenicillanic acid in aqueous solution or from hydrogenation of 6,6-dibromopenicillanic acid, is a powerful, irreversible, active-site-directed inhibitor of several typical beta-lactamases (penicillinase; penicillin amido-beta-lactamhydrolase, EC 3.5.2.6); 6-alpha-bromopenicillanic acid, being completely inhibited at less than micromolar concentrations through what is probably a 1:1 interaction. The B. licheniformis exoenzyme is similarly susceptible, while the Staphylococcus aureus enzyme and the Escherichia coli (R factor) enzyme are less so; the B. cereus beta-lactamase II is not inhibited. Very high concentrations (greater than or equal to 0.1 M) of benzylpenicillin, a good substrate, are required to significantly reduce the rate of inhibition of B. cereus beta-lactamase I by 6-beta-bromopenicillanic acid.


Mechanism of adenylate cyclase activation through the beta-adrenergic receptor: catecholamine-induced displacement of bound GDP by GTP. The fate of the guanyl nucleotide bound to the regulatory site of adenylate cyclase was studied on a preparation of turkey erythrocyte membranes that was incubated with [3H]GTP plus isoproterenol and subsequently washed to remove hormone and free guanyl nucleotide. Further incubation of this preparation in the presence of beta-adrenergic agonists resulted in the release from the membrane of tritiated nucleotide, identified as [3H]GDP. The catecholamine-induced release of [3H]GDP was increased 2 to 3 times in the presence of the unlabeled guanyl nucleotides GTP, guanosine 5'-(beta,gamma-imino)triphosphate [gpp(NH)p], GDP, and GMP, whereas adenine nucleotides had little effect. In the presence of Gpp(NH)p, isoproterenol induced the release of [3H]GDP and the activation of adenylate cyclase, both effects following similar time courses. The findings indicate that the inactive adenylate cyclase possesses tightly bound (GDP, produced by the hydrolysis of GTP at the regulatory site. The hormone stimulates adenylate cyclase activity by inducing an "opening" of the guanyl nucleotide site, resulting in dissociation of the bound GDP and binding of the activating guanosine triphosphate.


Structural markers on core protein p30 of murine leukemia virus: functional correlation with Fv-1 tropism. Tryptic peptide maps from more than 50 isolates of murine leukemia virus (MuLV) have shown that, in general, the structure of core protein p30 is highly conserved. However, a structurally variable region of p30 has been identified that is functionally associated with Fv-1 tropism. On the basis of this structural variability, MuLV strains can be classified as B-tropic, N-tropic, xenotropic, and/or as being derived from wild mice. Certain xenotropic viruses have a p30 like that of B-tropic MuLV and presumably would be subject to restriction in cells containing an Fv-In allele. Other p30 structural markers serve to distinguish the exogenous Friend, Moloney, and Rauscher viruses from endogenous MuLV. Furthermore, some MuLV strains have structural differences in their p30s that are useful as strain-specific markers. Finally, a possible sarcoma-associated alteration in the structure of p30 has been noted in the ml clone of Moloney murine sarcoma virus.


Mechanism of the in vitro breakdown of guanosine 5'-diphosphate 3'-diphosphate in Escherichia coli. Degradation of guanosine tetraphosphate (ppGpp) involves an enzyme associated with the ribosomal fraction from spoT+ strains of Escherichia coli. Double-label experiments with pp[3h]gpp, pp[3H]Gpp, or pp[3H]Gpp as substrate strongly suggest that ppG is the degradation product and that the enzyme releases two phosphates coordinately from the 3' position of ppGpp. In the absence of pppA this reaction proceeds in an uncoupled fashion, yielding ppG and PPi, but in the presence of pppA the decay is considerably enhanced and a pppA-ppi exchange reaction occurs in which the 3'-pyrophosphoryl group of ppGpp displaces the gamma and beta phosphates of pppA. Sodium PPi at 4 m7 inhibits decay of ppGpp regardless of whether or not pppA is present.


Changes in hepatic messenger RNA for phosphoenolpyruvate carboxykinase (GTP) during development. Phosphoenolpyruvate carboxykinase (GTP) [GTP;oxaloacetate carboxy-lyase(transphosphorylating); EC 4.1.1.32] is absent in rat liver cytosol during fetal life and is synthesized initially at birth. De novo synthesis of the enzyme can be induced prematurely by injection of dibutyryl cyclic AMP or glucagon into fetal animals in utero. In this study a wheat germ translation assay was used to quantitate the level of total functional mRNA for phosphoenolpyruvate carboxykinase in the liver of fetal rats at 21 days of pregnancy under different induction situations. The translatable mRNA for the enzyme was marginally detectable in fetal rat liver. Administration of either glucagon or dibutyryl cyclic AMP to fetal rats in utero caused a marked induction of functional mRNA for this enzyme. Three hours after administration of dibutyryl cyclic AMP, the level of translatable mRNA increased almost 23-fold, but by 6 hr the level dropped approximately 60%. Administration of actinomycin D prior to dibutyryl cyclic AMP in 21-day fetal rats prevented the appearance of newly synthesized poly(A)-containing RNA in the cytoplasm as well as the induction of translatable mRNA for phosphoenolpyruvate carboxykinase. In animals delivered prematurely and maintained for varying periods, the translatable mRNA for the enzyme accumulated in the liver at a rate comparable to that observed for enzyme synthesis.


Transfer of duck cell DNA sequences to the nucleus of 3T3 cells by Rous sarcoma virus. Peking duck cell nuclear DNA has no complementarity to RNA of Prague C Rous sarcoma virus (RSV). Upon infection of Peking duck cells by Prague C RSV, polynucleotide sequences complementary to Peking duck cell nuclear DNA can be detected in the high molecular weight RNA from progeny Peking duck cell passaged RSV (RSVPD) by RNA.DNA molecular hybridazation. When 3T3 cells are subsequently infected by RSVPD, polynucleotide sequences complementary to Peking duck cell nuclear DNA can be detected in the 3T3 cell nuclear DNA by RNA.DNA molecular hybridization. The potential consequences of the transfer of the Peking duck cell nuclear DNA from the avian to the murine cells are discussed.


Temperature-sensitive replication of H5ts125 adenovirus DNA in vitro. A soluble adenovirus DNA replication system has been prepared in extracts of HeLa cells infected with the temperature-sensitive mutant H5ts125. These nuclear extracts synthesize full-sized viral DNA at 30 degrees but are temperature sensitive in this function at 38 degrees. A 72,000-dalton DNA-binding protein purified from cells infected with wild-type virus complements the temperature-sensitive defect and restores viral DNA replication to 74% of normal values.


Inhibition of mammalian protein kinase and phosphodiesterase activities by a cyclic AMP-like compound isolated from higher plants. A cyclic AMP-like substance has been isolated from higher plant tissues which can be quantitated with the use of a radioimmunoassay similar to that described by A. L. Steiner, D. M. Kipnis, R. Utiger, and C. Parker [(1969) Proc. Natl. Acad. Sci. USA 64, 367-373]. This compound has been extensively purified and is chromatographically distinct from authentic cyclic AMP. This cyclic AMP-like compound inhibited beef heart 3':5'-cyclic-nucleotide phosphodietsterase (3':5'-cyclic-nucleotide 5'-nucleotidohydrolase, EC 3.1.4.17), with half-maximal inhibition occurring at a concentration of 7.6 X 10(-10) M cyclic AMP equivalents. The compound also inhibited cyclic AMP-dependent protein kinase (ATP:protein phosphotransferase; EC 2.7.1.37) from bovine heart, with half-maximal inhibition of mixed histone phosphorylation occurring at 8.0 X 10(-11) M cyclic AMP equivalents. Equipotent inhibition of phosphorylation and associated trace ATPase activity were observed with the purified catalytic subunit of cyclic AMP-dependent protein kinase from calf thymus with a synthetic heptapeptide as substrate. Moreover, steady-state kinetic analysis of this inhibition in the latter system showed it to be nonlinear and noncompetitive versus MgATP.


Isolation, characterization, and synthesis of a corticotropin-inhibiting peptide from human pituitary glands. A corticotropin-inhibiting peptide (CIP) has been isolated from human pituitary extracts. It consists of 32 amino acids with a proposed sequence identical to residues 7--38 of corticotropin (ACTH). The peptide has been synthesized by the solid-phase method. The melanotropic activity of the peptide is estimated to be 30% of the potency of ACTH. It is devoid of corticosteroidogenic activity but is able to inhibit ACTH-stimulated corticosterone production in isolated rat adrenal cells.


Immobilized lipid in acetylcholine receptor-rich membranes from Torpedo marmorata. The lipid environment of acetylcholine receptor-rich membranes from Torpedo marmorata has been studied with spin labels. The electron spin resonance spectra of both stearic acid and steroid probes in the membranes revealed an immobilized lipid component, in addition to the fluid component which is found in aqueous bilayer dispersions of the extracted lipids. The spin labels also cause a differential paramagnetic quenching of the intrinsic protein fluorescence of the membranes, which is sensitive to the action of cholinergic ligands and follows a modified Stern-Volmer law. Electron spin resonance difference spectroscopy shows that the protein-associated lipid is immobilized with respect to rotation both around and perpendicular to the long molecular axis, with correlation times : formula: (see text) approximately 50-70 ns. The proportion of lipid in the immobilized component is greater than calculated for a single boundary layer around the protein and corresponds more closely to the total interstitial lipid occupying the area between densely packed protein units in acetylcholine receptor-rich membranes.


Rat oocyte maturation in vitro: relief of cyclic AMP inhibition by gonadotropins. The hormone-independent, spontaneous maturation that rat oocytes undergo in vitro can be inhibited by derivatives of cyclic AMP and inhibitors of cyclic nucleotide phosphodiesterase. In this study, we have shown that this inhibition of maturation can be partially relieved by preparations of ovine and rat luteinizing hormone or follicle-stimulating hormone. The ability of gonadotropins to foster the resumption of maturation in cultures of cyclic AMP-inhibited oocytes suggests that this system is suitable for studies of the hormonal control of oocyte development. The dose and time dependency of the response to gonadotropins has been examined in order to study the role of these hormones in oocyte maturation and to compare this effect to other known responses of the cumulus-oocyte complex. These studies show that highly purified preparations of rat gonadotropins are less effective inducers of maturation than the more commonly used, but considerably less purified, preparations of ovine gonadotropins. Almost complete relief of inhibition is observed, however, when the oocytes are exposed to a combination of rat luteinizing hormone and follicle-stimulating hormone. Oocyte maturation was not influenced by the sex steroids progesterone or 17beta-estradiol. Our results suggest that: (i) cyclic AMP is involved in the intrafollicular inhibition of oocyte maturation; (ii) both gonadotropins are required for maximal stimulation of the resumption of oocyte meiosis; and (iii) steroids are not involved in this response to gonadotropins.


Simian virus 40 A gene function: DNA content analysis of Chinese hamster cells transformed by an early temperature-sensitive virus mutant. Replication of two Chinese hamster embryo cell lines transformed by an early temperature-sensitive mutant of simian virus 40, tsA58, was examined by flow microfluorometry and autoradiography of [3H]thymidine-labeled cells in order to determine whether transformed cell DNA synthesis is initiated by the virus A gene. At the permissive temperature (37 degrees), cells transformed by the mutant were like the wild-type virus transformants in appearance, colony-forming ability, high saturation density, and rapid replication. At the nonpermissive temperature (40.5 degrees), the tsA58 transformed cells resembled normal embryo fibroblasts and seem to return to normal growth patterns. Although both mutant transformed cell lines at 40.5 degrees appeared to cease growth at low saturation density, the cells did not enter a resting state, but continued to replicate. The cultures were maintained at low densities by a balance among cell replication, cell death, and sloughing of dead cells into the supernatant. These results suggest that the simian virus 40 A gene function effected by the tsA58 mutation does not prevent Chinese hamster embryo transformed cells from entering a resting state, although the gene may control other phenotypic characteristics of transformation.


Processing of 60,000-dalton sarc gene protein synthesized by cell-free translation. In this report we show that antiserum prepared against the Mr60,000 transformation-specific antigen of Rous sarcoma virus immunoprecipitates both the Mr60,000 and Mr 25,000 transformation-specific proteins that are synthesized by cell-free translation of virion RNA; however, in the cell-free system the Mr 60,000 protein appears to be synthesized as a precursor that is approximately Mr 2000 larger than the [35S]-methionine-labeled protein immunoprecipitated from Rous sarcoma virus-infected cells. Peptide mapping of the cell-free translation product and of this cellular protein has confirmed that they are structurally related to one another. The addition of membrane vesicles to the reticulocyte lysate system during translation specifically cleaves a Mr 2000 segment from the Mr 60,000 protein so that it comigrates with the cellular species. Secretory proteins and probably at least some integral membrane proteins are synthesized with short hydrophobic signal sequences at their NH2 terminus. Two facts suggest that the segment lost from the Mr 60,000 transformation-specific protein is a signal sequence: (i) the membrane vesicles process the Mr 60,000 protein only during translation, and (ii) the processed protein is sequestered by the vesicles.


Regulation of corneal collagenase production: epithelial-stromal cell interactions. Mixtures of epithelial and stromal cells isolated from normal adult rabbit cornea, when cocultured in the presence of cytochalasin B, produced latent collagenase, whereas neither cell type alone, nor the mixture in the absence of this agent, did so. The enzyme, a characteristic animal collagenase, required proteolytic activation. The relative concentrations of epithelial and stromal cells had a profound effect on on collagenase production, the enzyme activity bieng directly proportional to the number of stromal cells but inversely proportional to the number of epithelial cells. The amount of enzyme released into the medium was also directly proportional to cytochalasin B concentration. Media conditioned by cytochalasin B-treated epithelial or stromal cells did not stimulate collagenase secretion by the other cell type. The data suggest direct cell contact or close proximity as the mode of productive interaction and tentatively identify the stromal cell as the source of enzyme and the epithelial cell as a stimulator.


Depression of cytidine 3':5'-cyclic monophosphate phosphodiesterase activity in developing tissues of guinea pigs. Changes in levels of the newly discovered cytidine 3':5'-cyclic monophosphate (cyclic CMP) phosphodiesterase in some representative tissues (cerebral cortex, kidney, intestine, liver, heart, and lung) of guinea pigs from various developmental stages (fetus, neonate, pup, and adult) were studied and compared with those of cyclic AMP and cyclic GMP phosphodiesterases in the same tissues. It was observed that the tissue levels of cyclic CMP phosphodiesterase were invariably the lowest in every one of the fetal tissues examined, the highest in the corresponding tissues from the pups and adult, with intermediate levels seen in some neonatal tissues. The patterns of the ontogenetic changes in levels of cyclic AMP and cyclic GMP phosphodiesterase activities, however, were variable and tissue specific. These findings suggest that the depressed cyclic CMP phosphodiesterase activity (hence, the elevated cyclic CMP concentration) is perhaps a common factor in developing tissues undergoing rapid cell proliferation. The data also suggest that metabolism of cyclic CMP is perhaps more closely related to cell proliferation than the metabolism of cyclic AMP or cyclic GMP.


Age-dependent relaxation of gene repression: increase of endogenous murine leukemia virus-related and globin-related RNA in brain and liver of mice. A progressive loss in the ability of cells to maintain their normal specialized states of differentiation could be the common denominator underlying much of the physiology and pathology associated with the mammalian aging process. We investigated this possibility by searching for an age-dependent derepression of endogenous genes in tissues in which they are not normally expected to be expressed. Complementary DNA (cDNA) probes to globin genes and to the murine leukemia type C RNA virus (MuLV) genome were used to detect the presence of RNA complementary to these genes in RNA extracted from brain and liver in young and old mice. Significant amounts of globin RNA were found in brain nuclei and cytoplasm. No age difference was found in the globin RNA sequences present, but the number of globin RNA molecules increased from about 15 in 6-month to 34 in 27-month-old animals for nuclei and 280 in 6-month to 500 in 27-month-old animals for cytoplasm. Similar results were found for liver. RNA complementary to the MuLV cDNA probe was also found but, in contrast to globin, the different RNA sequences increased in both brain and liver nuclei from about 45% of the MuLV genome in 6-month to 72% in 27-month-old animals. The number of MuLV-related RNA molecules remained constant at about 22 and 38 per nuclei for brain and liver, respectively. Derepression of genes of this magnitude could result in a time-dependent increase in virus-related diseases and a general deterioration of the organism.


Viable deletion mutants in the simian virus 40 early region. For the purpose of isolating hr-t-like mutants of simian virus 40, we have constructed variants that have lost the unique site for the restriction enzyme Taq I at 0.565. Five mutants have been isolated and characterized by restriction enzyme analysis. All of them produce a normal size T antigen. Four produce a t antigen reduced in size as well as in amount; the fifth one does not seem to make any t antigen at all. The ability of these mutants to transform mouse cells in vitro, as tested by anchorage dependence, is clearly altered; however, the defect is only partial. In the same test, the mutants can complement a tsA mutant for transformation and therefore define a second complementation group in the simian virus 40 early region.


Another chromosomal assignment for a simian virus 40 integration site in human cells. Somatic cell hybrids derived from fusion of GM637, a human cell line transformed by simian virus 40, and mouse B82 cells were examined for simian virus 40 T antigen, V antigen, and viral DNA. All hybrid cell lines that contained viral DNA were T-antigen positive. Cells that did not have viral DNA were T-antigen negative. We determined that there is a single viral insertion in these hybrid cells. Correlation of T-antigen expression and viral DNA with the partial complements of the human genome retained in the hybrids shwed that the inserted viral genome is in human chromosome 8. The integrated viral DNA is stable; free viral DNA found in GM637 does not insert at other potential sites in the human genome.


Selective association of murine T lymphoblastoid cell surface alloantigens with Mycoplasma hyorhinis. Mycoplasma hyorhinis, isolated by isopycnic centrifugation from supernatants of a persistently infected murine T lymphoblastoid cell line, demonstrated the presence of the Thy-1.1 differentiation alloantigen and H-2Kk histocompatibility antigens. The murine leukemia virus-related gp70 antigen also present on the surface of these lymphoblastoid cells was absent from mycoplasma preparations. Quantitative assessment of Thy-1.1 present in preparations of M. hyorhinis revealed a specific activity greater or equal to that of membrane preparations from lymphoblastoid cells, suggesting a marked accumulation of this T lymphoblastoid cells, suggesting a marked accumulation of this T lymphocyte antigen by membrane-associated mycoplasmas. The accumulation of the Thy-1.1 antigen in association with purified mycoplasmas was also demonstrated in lymphoblastoid cells experimentally infected with a defined culture of M. hyorhinis.


Characterization of a feline sarcoma virus-coded antigen (FOCMA-S) by radioimmunoassay. A radioimmunoassay has been developed that detects a unique antigen encoded by the genome of the feline sarcoma virus (FeSV). Pseudotype viral particles containing an FeSV-specific polyprotein (p85) were used both as a source of antigen and to prepare specific antisera in rabbits. Because p85 contains antigens related to two structural proteins (p15 and p12) of feline leukemia virus (FeLV), antibodies directed to these were adsorbed with purified FeLV proteins. The adsorbed rabbit antiserum bound to antigenic determinants (designated FOCMA-S) which are also present in p85 and reacted specifically in immunofluorescence tests with rat cells transformed by FeSV and with FOCMA-positive cat lymphoid tumor cells. Competition assays detect FOCMA-S in pseudotype type C viruses rescued from FeSV-transformed mink and rat cells but not in heterologous type C helper viruses or in FeLV. A crossreactive antigen was also detected in pseudotypes of Kirsten sarcoma virus. The assay permits the quantitative measurement of an FeSV-coded protein whose expression is associated with viral transformation.


Isolation of a herpesvirus-specific DNA polymerase from tissues of an American patient with Burkitt lymphoma. A DNA polymerase (DNA nucleotidyltransferase) has been partially purified from a neck mass of an American patient with Burkitt lymphoma and separated from the cellular DNA polymerases. The molecular weight of the enzyme was approximately 90,000. The enzyme differs from the cellular DNA polymerases, but resembles herpes-virus-induced DNA polymerase in its primer template preference, high monovalent cation requirement for activity, and sensitivity to phosphonoacetate. Enzyme activity was inhibited specifically by an antibody directed against herpes-simplex-virus-induced DNA polymerase but not by antibodies directed against DNA polymerase alpha of HeLa cells and DNA polymerase gamma of a normal human lymphoblast cell line, NC37. Although serum of the patient with Burkitt lymphoma contained high Epstein-Barr virus titer, addition of the serum to the assay mixture did not have any effect on the activity of Burkitt lymphoma DNA polymerase. Tissues from spleen and liver of the patient with Burkitt lymphoma did not contain the herpes-virus-induced DNA polymerase. Detection of the herpes virus polymerase in the Burkitt lymphoma tissue provides additional evidence for the association of Epstein-Barr virus with this malignancy.


Formation of high density lipoprotein2-like particles during lipolysis of very low density lipoproteins in vitro. The effects of lipolysis of human plasma very low density lipoprotein (VLDL) on the structure and composition of high density lipoproteins (HDL) have been investigated. Lipolysis was performed in a controlled system in vitro containing VLDL (d less than 1.006 g/ml) and HDL3 (d = 1.125-1.210 g/ml) from human plasma and lipoprotein lipase (EC 3.1.1.34) purified from bovine milk. Lipolysis of VLDL caused profound changes in HDL3. Protein, phospholipid, and cholesterol liberated from VLDL during its lipolysis were transferred to the HDL3 particles. As a consequence of this in vitro transfer, the chemical composition and biophysical properties of HDL3 were substantially altered. The newly formed particles exhibited a flotation rate (F01.20) of 6.7 and a hydrated density of 1.110 g/ml. The chemical composition closely resembled that of native HDL2, and their size was slightly larger than that of the precursor HDL3. When HDL3 and postlipolysis HDL2 were subjected to ultracentrifugation under flotation velocity and equilibrium conditions, both proved to be stable particles. These results, when extrapolated to in vivo conditions, suggest an important metabolic relationship between the levels of circulating VLDL and HDL2 in plasma. This relationship now permits a reasonable explanation for numerous in vivo observations in which the levels of VLDL and HDL2 change reciprocally.


A virus similar to human hepatitis B virus associated with hepatitis and hepatoma in woodchucks. Particles with properties similar to those associated with human hepatitis B were found in serum from woodchucks with chronic hepatitis and hepatocellular carcinoma. It is suggested that woodchuck hepatitis virus is a second member of a novel class of viruses represented by the human hepatitis B virus.


Endogenous RNA tumor viruses are activated during chemical induction of murine plasmacytomas. Plasmacytomas are induced in BALB/c mice by the intraperitoneal injection of pristane (2,6,10,14-tetra-methylpentadecane) after a latent period of six months and more [Anderson, P. N. & Potter, M. (1969) Nature 222, 994-995]. Spleen cells mesenteric lymph node cells, thoracic lymph node cells, and peritoneal exudate cells were prepared from pristane-treated and control uninjected BALB/c mice during the course of a 10-month period, and these cell suspensions were tested for the release of infectious murine leukemia viruses. Endogenous ecotropic and xenotropic murine leukemia viruses were expressed in pristane-treated mice during the latter part of the tumor induction period, in those cell populations in which transformed plasma cells appear, namely, peritoneal exudate cells and thoracic lymph node cells. The significance of preferential expression of both ecotropic and xenotropic murine leukemia virus in target cell populations following the administration of a carcinogen is discussed in terms of the possible formation of an oncogenic variant virus.


Serological characterization of B-tropic viruses of C57BL mice: possible origin by recombination of endogenous N-tropic and xenotropic viruses. The serological properties of the gag gene products p15 and p12 of N- and B-tropic viruses of C57BL mice have been examined. Although these viruses were serologically identical by competition assays for proteins gp71 and p30, they were readily distinguishable in competition assays for proteins p15 and p12. Two isolates of N-tropic viruses had p12s serologically indistinguishable from AKR murine leukemia virus p12, while two B-tropic isolates had distinctly different p12s. The latter p12s were serologically indistinguishable from the p12 purified from the B-tropic radiation leukemia virus (RadLV)/VL-3. Moreover, this p12 was indistinguishable from the p12 of the endogenous C57BL/Ka xenotropic virus. Similarly, the p15s of the B-tropic viruses were serologically distinct from the AKR murine leukemia virus type of p15, as was the p15 of one C57BL N-tropic virus, whilc another N-tropic isolate had a p15 identical to the AKR murine leukemia virus p15. These results are interprered to suggest that the endogenous N-tropic virus of C57BL mice undergoes recombination with the endogenous, xenotropic virus and that this mechanism is involved in the generation of B-tropic viruses in C57BL mice.


Activation of endogenous type C virus in BALB/c mouse cells by herpesvirus DNA. Several virion and nonvirion DNAs were tested for the ability to activate endogenous type C virus in BALB/c-derived mouse cells using the calcium precipitation technique. The DNAs from all herpesviruses tested activated xenotropic type C virus synthesis. These included DNAs from herpes simplex virus types 1 and 2, Epstein-Barr virus, human cytomegalovirus, SA8 virus, infectious bovine rhinotracheitis virus, pseudorabies virus, and herpes saimiri virus (M-DNA). In contrast, DNAs from vaccinia virus, simian virus 40, primate cells, bacteria, mycoplasma, and salmon sperm showed no ability to activate type C virus when tested under the same conditions. Several herpesviruses and vaccinia virus, which were highly infectious for the BALB/c cells used, were tested for their ability to activate type C virus after UV irradiation. All herpesviruses tested were positive, while vaccinia virus was negative. Unirradiated simian virus 40 also showed no ability to activate type C virus. Activation of type C virus by DNA from herpes simplex virus was observed after shearing or sonication of the DNA to an average size of 3 x 10(6) daltons, but was not observed with DNA sonicated to an average size of 1 x 10(6) daltons. Alkali denaturation of DNA from herpes simplex virus or treatment with DNase, but not RNase, destroyed its ability to activate type C virus, as did crosslinking of the DNA with 4,5',8-trimethylpsoralen (psoralen) and light.


Mouse brain deoxyglucose uptake after footshock, ACTH analogs, alpha-MSH, corticosterone or lysine vasopressin. The cerebral uptake of subcutaneously injected [3H]2-deoxy-D-glucose (2DG) in 16 brain regions was examined following 30 noncontingent random footshocks or the acute injection of saline, ACTH1-24 (0.5 microgram/g), ACTH/MSH4-10 (0.25 microgram/g), [D-Phe7]ACTH4-10 (0.25 microgram/g), [Met4SO2,D-Lys8,Phe9]ACTH4-9 (0.01 microgram/g), ALPHA-MSH (0.5 microgram/g), corticosterone (2.5 microgram/g) or lysine vasopressin (0.05 microgram/g). Footshock selectively decreased 2DG uptake in parietal cortex and brain stem, and increased that in the hypothalamus. Whole brain 2DG uptake was decreased by injection of saline or most of the hormones relative to uninjected animals, but this effect was probably peripheral since plasma glucose content was increased by the injections. The only regionally specific effect of the hormones was an increased 2DG uptake in olfactory bulb by saline, ACTH/MSH4-10 And corticosterone relative to uninjected animals. Since alpha-MSH had been reported previously to decrease blood flow (measured by antipyrene uptake) in all brain regions except occipital cortex [5,6], we directly compared antipyrene uptake with 2DG uptake in the same animals using a double-isotope procedure. The results revealed an increase in 2DG uptake relative to antipyrene in cortical regions relative to subcortical regions, contradicting earlier assumptions [19].


Involvement of brain cyclic AMP in the acute and chronic effects of morphine in the rat. Incubation of cerebral cortical slices of rat brain with 3H-adenosine in the presence of varying concentrations of morphine in vitro resulted in a dose-related increase in 3H-cAMP formation. In in vivo experiments, the rate of 3H-cAMP formation in cortical slices both 45 min and 4 h after the acute s.c. administration of a 10 mg/kg dose of morphine was significantly greater than that for saline-treated controls. A significant enhancement of cortical 3H-cAMP formation likewise became apparent 72 hr after s.c. implantation of two morphine pellets. This was not evident after only 24 h. In similar experiments undertaken with hypothalamic tissue, however, the rate of conversion of 3H-adenosine to 3H-cAMP remained similar to that of the controls. Administration of exogenous cAMP antagonized the analgesic response to morphine in both nontolerant and tolerant rats and accelerated the development of tolerance to morphine. These results may provide further evidence for a role of cAMP in the mediation of the central actions of morphine.


Levodopa-mediated alteration in the activity of pyridoxal kinase in rat basal ganglia. The acute intraperitoneal administration of 400 mg/kg of levodopa reduced the concentration of pyridoxal phosphate and increased the activity of pyridoxal kinase. The chronic oral administration of 100 mg/kg/day of levodopa did not alter the concentration of pyridoxal phosphate, but increased the activity of pyridoxal kinase in 7 days. This initial elevation in kinase activity returned to normal levels in 2 weeks, became reduced in 3 weeks and then fluctuated back to normal levels in 4 weeks. These results are interpreted to indicate that chronic intake of levodopa results in adaptive alteration in the activity of pyridoxal kinase. This maintains the steady-state concentration of pyridoxal phosphate in order to meet the levodopa-induced enhancement in decarboxylation reaction.


Tubular carcinoma of the breast. Mammographic appearance and pathological correlation. Xeromammographic and pathological findings in 17 cases of tubular carcinoma of the breast are presented. No patients had axillary lymph node involvement. The mean lesion size was 0.95 cm. Thirteen lesions (76%) were clinically occult. Because of the characteristic histological pattern of infiltrative growth and marked desmoplasia, 81% (13/16) were seen on xeromammography as scirrhous masses, 19% (3/16) only as microcalcifications. None appeared as a well-circumscribed mass or poorly defined density.


The grain in the stone: a computer search for hidden CT patterns. Relatively simple but time-consuming computer manipulations of basic CT images have been carried out. We have been unsuccessful in finding significant information when the skewness or kurtosis of the CT image is computed and displayed, but have found recognizable and useful new gray scale images upon reconstruction of sigma mode textures.


Effectiveness of a specially trained pharmacist in a rural community mental health center. Pharmacists having training in psychopharmacology and psychiatry are being specially trained to function in community mental health centers as a resource in psychopharmacology and to provide direct care to patients. Management information data over a 3-year period from eight rural mental health clinics indicate that these pharmacists can successfully maintain large numbers of stabilized psychiatric patients within their communities. The cost of a pharmacist's services is, conservatively, one-half that of a psychiatrist's services. A followup study of the stabilized and active aftercare outpatients in the 10-county rural area in which the 8 clinics are located revealed that those patients who received all their direct care from the pharmacist were functioning at a slightly healthier level than the other aftercare patients. The pharmacist's patients indicated that they were at least as satisfied with their care as were the aftercare patients who received care from other mental health professionals. If the results of this study can be generalized to other community mental health centers in rural areas, a pharmacist can provide services effectively when psychiatrists are inaccessible or unavailable or when funds for mental health professionals are limited.


Computed tomography in astrocytomas. A statistical analysis of the parameters of malignancy and the positive contrast-enhanced CT scan. Four histopathological indices of malignancy--cellularity, pleomorphism, vascularity, and necrosis--were correlated with the positive-contrast CT scan in 84 patients with pathologically proved astrocytoma. Vascularity and necrosis correlated best with the scan in supratentorial tumors, while vascularity and cellularity correlated significantly with the scan in cerebellar astrocytomas. These findings indicate that the degree of malignancy of a supratentorial astrocytoma can be inferred from the contrast-enhanced scan, but that this is not generally possible with posterior fossa tumors.


99mTc-pyrophosphate imaging in experimental mesenteric infarction: relationship of tracer uptake to the degree of ischemic injury. Four-hour balloon occlusion of the cranial mesenteric artery in 13 dogs produced a spectrum of widespread ischemic injury to the small bowel ranging from superficial mucosal infarction to transmural infarction. Twenty-four hours after the beginning of balloon occlusion, 15 mCl of 99mTc-pyrophosphate was injected into the cranial mesenteric artery in 8 dogs and into a peripheral vein in 5. Gamma camera imaging of the abdomen followed three hours later. Four of 8 dogs injected intra-arterially showed positive images with full-thickness mucosal or transmural infarction. The 4 intravenously injected dogs with positive images had transmural infarction. Only minimal ischemic injury with superficial mucosal infarction was found in the 5 dogs with negative images. Tracer uptake per gram infarcted tissue was highest after intra-arterial injection. It was concluded that 99mTc-pyrophosphate allows external imaging of extensive intestinal infarctions after both intra-arterial and intravenous injection.


Abnormalities resulting from intra-adnexal injection of glucose in the rabbit embryo--an experimental model of "amniotic disease". Intra-adnexal injections of glucose into the rabbit embryo have induced amputations of digits or segments of limbs, congenital grooves, amniotic bands, club feet, syndactyly, hare lip, anencephaly, and ulcerations of the scalp. We have thus reproduced all the anomalies which are encountered in the clinical syndrome of "amniotic disease." These anomalies result from destruction of the cutaneous epithelium and the subjacent mesenchymatous cells, and extravasation of blood with hematoma formation around the superficial vessels. This general mechanism explains most of these anomalies. Intra-adnexal injections of glucose thus constitute an external trauma for the embryo and is good experimental model of amniotic disease.


Prostaglandin I2 stimulation of granulosa cell cyclic AMP production. The ability of prostaglandin I2 (PGI2) to stimulate cyclic AMP production by granulosa cells, isolated from intact immature rats, has been demonstrated in vitro. The minimal effective dose was 15 ng/ml, which was comparable to the minimal effective dose for PGE2. However, a concentration of 15 microgram/ml PGI2 was required to stimulate cyclic AMP production maximally, compared to a concentration of 1 microgram/ml PGE2, which produced the maximum response. It therefore appears that PGI2 is not more effective than PGE2 in stimulating cyclic AMP production in granulosa cells, and is possibly less effective. Submaximal concentrations of PGI2 appeared to be able to modify the stimulation of cyclic AMP production by follicle-stimulating hormone (FSH), but whether or not PGI2 plays any role in follicular function remains to be established.


Cholesterol arachidonate as a prostaglandin precursor in adrenocortical cells. Rat adrenocortical cells were incubated with labeled arachidonate, and the radioactivity in unesterified fatty acids was reduced by washing with 2% albumin solutions. These cells were then incubated for two hours in the absence and presence of 7.1 x 10(-10)M ACTH. During subsequent incubation of prelabeled cells with ACTH, both the mass and radioactivity of arachidonate in adrenocortical cholesteryl esters was depleted to the same extent (30--32%). The released arachidonate was in part incorporated into phospholipids, and there was also a significant increase in unesterified arachidonic acid. During this period, there was also increased incorporation of arachidonate into labeled prostaglandins. Of this increase, 92% by isotope analysis, and 88% by radioimmunoassay techniques was attributable to prostaglandins of the E pathway. These data demonstrate that prostaglandin E synthesis is specifically increased during ACTH stimulation of rat adrenocortical cells and suggest that a major source of the arachidonate substrate for this synthesis is derived from hormone-dependent hydrolysis of cortical cholesteryl esters.


Interaction of prostaglandins and adenosine 5'-triphosphate in the noncholinergic neurotransmission in rabbit detrusor. Part of the excitatory transmission in rabbit detrusor is noncholinergic and nonadrenergic, and prostaglandins (PGs) and adenosine 5'-triphosphate (ATP) have been implicated in this transmission. The present experiments investigate the possibility of an interaction between PGs and ATP in rabbit detrusor. Indomethacin (2.8 muM) depressed the contraction produced by ATP although it did not antagonize the contraction produced by carbachol. Treatment of detrusor strips with 1.5 mM ATP depressed the frequency response curve in field stimulated tissues. This depression was additive with that produced by atropine. In the present experiments indomethacin did not significantly augment the effect of desensitization with ATP. It is suggested that the atropine-resistant neurotransmission in rabbit detrusor may involve both ATP and PGs acting in cooperation.


Role of antitumor antibiotics in current oncologic practice. The antitumor antibiotics have thus made a major impact on oncologic practice. The continued search for productive strains of these organisms should be encouraged. In addition, the activity and toxicity spectrum suggests the need for vigorous analog development. An active anthracycline devoid of cardiotoxicity; a bleomycin with no effect on pulmonary tissue; an analog of streptozotocin devoid of nephrotoxicity; these would be advances of inestimable benefit to the cancer patient of the future.


Observations on the neuromuscular blocking action of alcuronium in the dog and its reversal by neostigmine. The action of alcuronium chloride on neuromuscular transmission in the dogs was investigated by electrical and mechanical methods. The mean duration of action was 70 min. Reversal of its action was produced with atropine and neostigmine.


The resistance to reinfection of tracheal organ cultures from chickens previously infected with avian infections bronchitis virus. Lower titres of avian infectious bronchitis (AIB) virus were found following the infection of tracheal organ cultures prepared from chickens that had been given AIB virus intranasally six weeks previously than were found following the infection of organ cultures prepared from untreated or from passively-immune chickens. No infectious virus was found in the tracheal organ cultures at the time they were prepared.


The experimental infection of piglets with a porcine reovirus. A strain (Quebec) of reovirus isolated from the faeces of a pig with dysentery was neutralised by reovirus type 1 antiserum. Four of eight hysterectomy-produced, colostrum-deprived (HPCD) piglets dosed orally with the third cell culture passage of the virus developed diarrhoea and showed focal areas of villous atrophy in the small intestine. The virus was isolated from the intestinal tract of all eight specific pathogen free piglets, but not from three control animals. Nine germ-free piglets dosed orally with the eight cell culture passage of the virus showed neither clinical signs nor lesions, but virus was recovered from their intestinal tracts for 14 days after infection. No virus was isolated from four control germ-free piglets.


Scanning electron microscope study of Sendai virus fusion between HeLa cells and chick erythrocytes. Chick erythrocytes were fused with HeLa cells by Sendai virus and preparations examined by scanning electron microscopy at different times after fusion. Heterokaryons were usually formed by fusion of erythrocyte ghosts with HeLa cells. Occasionally whole erythrocytes were engulfed but there was no evidence that free nuclei fused. Initial inter-cell attachments were usually, and possibly always, made at the site of an attached virus particle. This study helps to correlate topographical findings with previous two-dimensional studies with the transmission electron microscope and may also provide a model system for the fusion of parasitised erythrocytes with eukaryotic cells.


The recruitment times and firing patterns of the medullary respiratory neurones of the cat. The brainstem of anaesthetized cats was explored for respiratory neurones with microelectrodes in the regions of the nucleus retroambigualis (ventrolateral region) and the nucleus tractus solitarius (dorsomedial region). These neurones were analysed with respect to their firing patterns and recruitment times, as referenced to the respiratory cycle. All of the respiratory neurones showed a stable and ordered pattern of firing. Four groups of neurones with similar characteristics within each group, but differing from each other, were statistically examined; the inspiratory neurones of the dorsomedial region, the 'late' inspiratory neurones of the ventrolateral region, the 'early' inspiratory neurones of the ventrolateral region and the expiratory neurones of the ventrolateral region.


[Clinical studies of the role of the fluorescent circulating antibody test in the diagnosis of active tuberculosis]. The value was studied, of the test of circulating fluorescent antibodies against M. tuberculosis in establishing the diagnosis of active tuberculosis in 610 patients classified in 4 groups: 1. active tuberculosis bacteriologically confirmed; 2. active tuberculosis with negative bacteriological examination, but confirmed by other methods of criteria; 3. stabilized tuberculosis; 4. non-tuberculous affections. The specificity of the test in active tuberculosis was of 80% in adults and of 91,5% in children. The test was negative in 80,3% of stabilized tuberculosis cases and in 69% of non-tuberculous affections. The sensitivity was lower in active tuberculosis with positive bacilloscopy (67%) against active tuberculosis with negative bacilloscopy (90,6%), a fact explained by a high consumption of anti-bacillary antibodies, demonstrated by positivation of the test when bacilloscopy becomes negative. The use of the test is indicated in establishing a therapeutic test, and in the context of clinical and radiologic explorations, as well as in the expertising of working capacities of previous tuberculosis patients.


[Studies of resumption of treatment with rifampin and ethambutol in patients with recurrent pulmonary tuberculosis]. The immediate and the late results are analyzed, of resumption of treatment with RMP and EMB in 50 patients with relapsing pulmonary tuberculosis that had not been treated with these drugs previously. After the first year of treatment negativation of the sputum by microscopy and culture was obtained in 45 of the patients, while 5 remained still positive. In the following two years 7 of the negative patients became positive again and finally 8 were solved completely and 4 became chronically ill. The most important of the unfavourable factors were simultaneous radiological and bacteriological relapses and chronic alcoholism. There was no primary or secondary resistance against the drugs used in the treatment. Intolerance to the drugs, although frequent, did not significantly affect the course of the treatment, especially because it occurred only at a later time.


Serologic markers of hepatitis A and B in chronic active hepatitis. Sera from 44 patients with a well-documented diagnosis of chronic active hepatitis (CAH) were analysed for antibodies to hepatitis A virus (anti-HAV), hepatitis B surface antigen (HBsAg) and anti-HBs, e-antigen (HBeAg) and anti-HBe, as well as antibodies to hepatitis B core antigen (anti-HBc). Twenty-two patients had serologic evidence of hepatitis A infection. The frequency of anti-HAV was low in patients under 50 years of age (21%) but high among older patients (72%). There was, however, no significant difference between patients and age-matched controls regarding the prevalence of anti-HAV in serum. Markers for hepatitis B virus were found in 10 patients or 23% as compared with about 10% in Swedish blood donors. The results indicate that hepatitis A virus is of little importance in the pathogenesis of CAH and confirm the association between hepatitis B virus and development of chronic active hepatitis.


Human leucocyte migration inhibitory factor (LIF). V. Specific regulation of the lymphokine serine protease activity by 3',5'-cGMP and, possibly, 2',3'-cCMP. The cyclic nucleotides guanosine 3',5'-cyclic monophosphoric acid (3',5'-cGMP) and cytidine 2',3'-cyclic monophosphoric acid (2',3'-cCMP) but not cyclic phosphodiesters derived from the bases adenine and uracil preserved LIF activity against the blocking effect of the serine protease inhibitor phenylmethylsulphonyl fluoride (PMSF). Phosphomonoesters derived from guanosine and cytidine as well as 2',3'-cGMP and 3',5'-cCMP were all inactive, indicating specificity for phosphodiester bonds and their respective positions in the two active nucleotides. The protection afforded by 3',5'-cGMP and 2',3'-cCMP was dose dependent. Thus, using 10(-3) M PMSF, 3',5'-cGMP was active at concentrations higher than 10(-5) to 10(-4) M, and 2',3'-cCMP at concentrations higher than 3 X 10(-4) to 10(-3) M. The more pronounced LIF-inhibitory effect obtained by increased concentrations of PMSF could be overcome by raising the levels of the nucleotides, indicating that the interactions between PMSF and the nucleotides with LIF were mutally exclusive. The possibility that 3',5'-cGMP and perhaps 2',3'-cCMP function as modulators of LIF is discussed, and models for the function of this lymphokine are proposed.


99mTc-pyrophosphate kinetics in man (relationship between bone uptake and mineral content). Both the kinetics and the regional distribution of 99mTc-PPi were studied in the skeleton of healthy subjects of various ages by surface counting using a scintillation counter placed over different skeletal areas. The results were compared with bone mineral content as determined by the photonic absorption technique. Kinetic data of bone uptake were obtained by subtracting the counts measured in blood from the counts recorded over each body surface after making proportional corrections. The surface-counting patterns show a higher generalized 99mTc-PPi bone uptake in children and adolescents than in adults. The relative regional distribution of the tracer shows no difference between the two groups. The tracer accumulated more rapidly in spongy bone than in compact bone, and the fixation process was carried out during the first 30 min for both spongy and compact bone. In youths, however, higher uptake values corresponded to lower values of mineral content. The findings enable the normal surface-counting pattern to be constructed and confirm that the 99mTc-PPi uptake is related to bone metabolism; they also suggest the application of external measurements for the study of metabolic bone diseases.


Effect of cyclic nucleotides, isoproterenol and cholera toxin on DNA synthesis triggered by mitogens. The effect of cyclic nucleotides, isoproterenol and cholera toxin on phytohaemagglutinin (PHA), concanavalin A (Con A) and anti-allotype-induced rabbit lymphoid cell proliferation was examined. Cholera toxin in concentrations ranging from 10(-8) microgram to 1 microgram per culture inhibited DNA synthesis, triggered by PHA, Con A and nocardia water-soluble mitogen (NWSM). It had the opposite effect on stimulation with antibodies directed against allotypic specificities of the immunoglobulin light chains: over the entire range of tested concentrations, cholera toxin stimulated DNA synthesis triggered by antibodies to Ab4, Ab5, Ab9 and to Aa1 allotypic specificities. Relatively high concentrations of dibutyryl adenosine 3':5'-cyclic monophosphate (cAMP) and isoproterenol (10(-3) M) inhibited mitogen-stimulated thymidine incorporation; lower concentrations (10(-9) M) had an enhancing effect. A similar enhancing effect was observed when high (10(-3) M) concentrations of dibutyryl guanosine 3':5'-cyclic monophosphate (cGMP) were used.


Effects of some connective-tissue active drugs on protocollagen proline hydroxylase activity. The effect on protocollagen proline hydroxylase of drugs reported to be capable of inducing the lupus erythematosus syndrome and in some way acting on connective tissue and inflammation was studied in an in vitro system for the hydroxylation of 14C-Pro-labelled protocollagen. Some derivatives were also studied. It was found that benzoic acid and phenothiazine derivatives, hydroxyphenols, (+)catechin, beta-amino propionitrile, certain B vitamins, dihydrazinophthalazine, cysteine and dimethylcysteine were inhibitors of PPH. The chelation of Fe2+ions by the compounds mentioned is suggested to be essential.


Effect of occupational exposure to organophosphorus insecticides on neuromuscular function. Neurophysiological investigations and determinations of cholinesterase activity on plasma and erythrocytes were carried out on 11 Swedish spraymen exposed to bromophos, diazinon, dursbane, and malathion. Plasma cholinesterase activity was significantly reduced after work, while erythrocyte cholinesterase activity was unchanged. In none of the workers with a decreased plasma cholinesterase activity after work could any related acute neuromuscular disturbance be detected when the men were tested with repetitive nerve stimulation and with single fiber electromyography. Signs of subclinical neuropathy were present as a slight reduction in sensory conduction velocity and increased fiber density in some workers.


[Immunosuppression as a concomitant effect]. 1. It is becoming increasingly clear that besides traditional immunosuppressive agents, an array of miscellaneous influences and conditions, as well as drugs used for other purposes, can affect the immune system. The potential pathogenetic role of immunity in an ever growing number of diseases, including infections, cancer, and autoimmune processes, confers on the concept of concomitant immunosuppression or immunotoxicology its due place in the discipline of immunopharmacology. 2. Microorganisms such as viruses, bacteria, and protozoa (and their products), as well as parasites, may depress immune reactivity. The mechanism of action, dose-response dependency, effect of timing, and relation to bacterial adjuvanticity need more thorough exploration. 3. The efficacy of immunosuppressive drug protocols can be improved by combining immunosuppressants without bone marrow toxicity, such as niridazole or free fatty acids, with the standard antiproliferative immunosuppressive agents. 4. Prostaglandin is a likely candidate for an essential role as an immunosuppressive effector. Its release from T-suppressor cells is triggered for example by microorganisms. Moreover, dietetic and metabolic factors, such as free fatty acids, and nervous impulses affect prostaglandin formation. If thus constitutes a link between the nervous system and, on the other hand, immunity and possibly cancer.


[Practical value of new knowledge on the structure and function of pituitary adenomas]. The usual classification of pituitary adenomas into eosinophilic, basophilic, and chromophobe types is no longer sufficient to correlate the histologic findings with the increasing number of clinical syndromes described in recent years. Some histologic forms are observed in several clinical syndromes, whereas some clinical entities may be caused by several histologic types. A new classification is mandatory. Recent endocrine, ultrastructural and histo-immunologic research has shown that the endocrine inactive adenoma, which has usually been related to the "chromophobe" adenoma, represents only a small group of the pituitary neoplasms. A more critical evaluation shows that 3 out of 4 patients present clinical or endocrine signs and findings of increased hormone secretion. Several authors have therefore suggested that pituitary adenomas be classified according to their secretory characteristics, which can be determined either by clinical and endocrine examination or by immunohistology and electron microscopy. Abnormally increased hormone secretion allows early detection of the neoplasm when the remaining normal gland is still able to recover its normal function and the endocrine symptoms therefore are fully reversible. The therapy of pituitary adenomas must normalize the hormone hypersecretion without causing new endocrine deficits, and must at the same time reverse neurological compression symptoms. The treatment may be either surgical excision, irradiation, medical inhibition of abnormal secretion, substitution of insufficient hormones or a combination of these methods. The prognosis of the endocrine symptoms is significantly better in small adenomas and full restoration of sexual function is even observed in an increasing number of patients.


[Urinary excretion kinetics of enzymes and proteins in the application of therapeutic doses of gentamycin]. 9 patients with chronic pyelonephritis were given gentamicin intramuscularly in an appropriate dosage for a period of 10 days. The urinary excretion of cells, protein, alanine aminopeptidase (EC 3.4.11.2),N-acetylglucosaminidase(EC 3.2.1.30) and alpha-galactosidase (EC 3.2.1.23) was determined daily. The most striking enzyme increase occurred in alanine aminopeptidase. The enzyme and protein output appears to be rhythmic and discontinuous. A 2--3-day rhythm is suggested. Rhythmic alterations of the digestion activity of the lysosomes are assumed to be the cause of these changes.


[A case of poliomyelitis type 1 in Switzerland in a non-vaccinated child]. A case of paralytic poliomyelitis in an unvaccinated child is reported. Besides clinical aspects, special interest is focused on immunological and virological findings. Paralytic disease in temporal connection with poliomyelitis vaccination is also discussed.


Carcinogenicity testing of chemicals with particular reference to organochlorine pesticides. The organochlorine chemicals comprise a large number of pesticides that are used widely throughout the world. The organochlorine pesticides given in the diet to mice are carcinogenic for the liver. They induce not only carcinomas of the liver, particularly at the higher doses, but also carcinomas and sarcomas in other organs in rats. They cause acute and chronic liver and kidney injury, which interferes with the development of carcinomas and sarcomas in rats. The testing of chemicals for carcinogenicity, with particular reference to organochlorine pesticides, includes discussions on the following topics: classification of hepatic lesions in mice and rats, toxicity versus carcinogenicity in the testing of chemicals, a comparison of carcinomas and cirrhosis of the liver in experimental animals and humans, and the significance of laboratory carcinogenicity findings to human health.


Invertebrate collagens.

Invertebrate collagens. The collagens of all major invertebrate phyla have been studied, but characterization has been thorough in only a few classes and in no case in the detail (such as sequence analysis) known for vertebrate collagen. Biochemical data on insect collagen are particularly sparse. Invertebrate and vertebrate collagens are strikingly similar, with some notably unique features in annelids and nematodes. Present data do not support the suggestion that invertebrate collagens resemble vertebrate basement membrane collagen. In invertebrates, as in vertebrates, collagens of specific tissues show differenes that probably reflect individual tissue requirements.


Silicosis in kaolin workers and firebrick makers. Nine patients with disabling pneumoconiosis were selected over a three-year period for study because of a work history of kaolin exposure. All had worked in the Missouri firebrick industry. Chest roentgenograms from all nine and lung tissue from two  were consistent with silicosis. Worksite analysis of airborne dust at several Missouri firebrick factories revealed that some current workers are exposed to both free silica (up to 4.5%) and cristobalite (up to 8.9%) at levels exceeding the threshold limit value (TLV). Analysis of airborne dust from several kaolin processing plants in South Carolina and Georgia revealed that a few workers are exposed to about 1% free silica at levels occasionally exceeding the TVL. We conclude that silicosis is a serious risk for Missouri kaolin workers and a potential risk for workers in South Carolina and Georgia. A prevalence survey is needed in Missouri to assess the problem.


Initial characterization of Trypanosoma lewisi surface membrane antigen. The purified T. lewisi were subjected to hypotonic lysis plus freezing and thawing in acetone dry ice bath. The trypanosome ghosts were obtained after repeated washing and centrifugation. The homogenized ghost suspension was assayed for enzyme Na++K+ ATPase activity to ratify the presence of the trypanosome surface membrane. Membrane solubilized in sodium dodecyl sulfate were fractionated by gel filtration on Sephadex columns equilibrated with the detergent and electrophoresis in polyacrylamide gel. Crude trypanosome surface membrane antigens were tested for their immunogenicity, administered to rats in Fraund's complete adjuvant. The results of these experiments indicated that the protective immunogen is tightly bound to the membrane since the use of strong anionic detergent is necessary in its extraction.


Prospects of vaccines against cancer in man. Preventive immunization is the cheapest and most effective approach to the management of epidemic disease. Certain forms of cancer, including Burkitt's lymphoma, carcinoma of the cervix, primary liver cancer, acute leukaemia and breast cancer show many characteristics of epidemic disease, and the possibility of producing preventive vaccine against these conditions merits serious consideration.


Glomus jugulare tumour in a dog; a case report. Unilateral ataxia of the head and trunk and right-sided paralysis of the trigeminal, facial, and hypoglossal nerves were the major neurologic dysfunctions in a nine-year-old male French bulldog. These symptoms together with the results of radiologic examination and bone scintigraphy pointed to a cranial base tumour on the right side. The tumour was histologically identified as a glomus jugulare tumour. A tumour of the right carotid body and a seminoma in the right testicle were additional findings at autopsy.


The lipid-chemical features of the metastatic tissues into the liver from the human gastric cancer, large intestinal cancer and malignant insulinoma. In order to clarify the biochemical features of metastatic tissues into the liver of human cancerous cells, 12 of primary cancerous tissues and 3 of metastatic tissues of the large intestinal cancer, 6 of primary cancerous tissues and 2 of metastatic tissues of the gastric cancer, and 3 of primary cancerous tissues and 3 of metastatic tissues of malignant insulinoma were studied lipid-chemically. Cancerous tissues and metastatic tissues into the liver were collected by biopsy or surgical operation. From each tissue, the total lipid was extracted and one part of the total lipid was separated into phospholipid and triglyceride by TLC. Then, the fatty acid composition and the fatty acid content of each lipid fraction were measured by GLC. The most remarkable findings were recognized in the phospholipid fatty acid composition of the tissues. Namely, the percentage values of C14:0 and C16:1 were larger and that of C20:4 was smaller in metastatic tissues than those of gastric primary lesions. As for the large intestinal cancer, the percentage value of C18:1 was smaller and that of C18:2 larger in metastatic tissues than those of primary lesions. In the malignant insulinoma, the percentage value of C18:0 was larger in metastatic tissues than that of the primary lesions.


Action of dihydroaflatoxins on rat liver mitochondria. 1. Membrane-dependent osmotic and redox effects of aflatoxins B2 and G2. Membrane-dependent osmotic and redox effects of two dihydroaflatoxins (aflatoxin B2 and G2) on isolated rat liver mitochondria were investigated. These toxins did not cause any marked swelling, inhibit any ATP induced contraction or stimulate ATPase activity of mitochondria at concentrations within the range 1--2 x 10(-6) M. Redox measurements on the respiratory chain showed that both toxins inhibited respiration between cytochrome c and the terminal oxygen. The significance of these findings in relation to the differential toxicity of the aflatoxins is discussed.


Estrogen synthetase in choriocarcinoma cell culture. Stimulation by dibutyryl cyclic adenosine monophosphate and theophylline. The stimulation of estrogen biosynthesis by N6,O2'--dibutyryl adenosine 3':5'-cyclic monophosphate and theophylline dbT) in cultures of the JAr line of choriocarcinoma cells was investigated by measuring the specific activity and kinetic constants of estrogen synthetase (aromatase) in the various subcellular fractions after differential centrifugation of homogenized cells in isotonic sucrose. The low speed (900xg) pellet, from cells grown with or without dbT and homogenized in isotonic sucrose, contains the majority of the aromatase activity and the highest aromatase specific activity. The aromatase specific activity in the homogenate of cells grown with dbT and in the various subcellular fractions is 4- to 10-fold higher than in cells grown without dbT. The Vmax of androstenedione (4-androstene-3,17-dione) aromatization in homogenates from dbT-stimulated cells (6.9 pmol estrogen/min per mg protein) is significantly increased over that measured in the absence of dbT (1.5 pmol estrogen/min per mg protein); the Km values, however, are not significantly different (average of 43.8nM in dbT-stimulated fractions; 53.2nM in control fractions). These results suggest that the increased aromatase specific activity in dbT-stimulated cells results from an increase in amount of active enzyme, rather than from an increase in affinity of the enzyme for its substrate.


Malignant fibrous histiocytoma of spermatic cord. Malignant fibrous histiocytomas are very rare, particularly those related to urogenital organs. We report herein the third known case of this entity involving the spermatic cord. Summary of the previously reported 2 cases is given with recent follow-up. Prognosis is poor.


Enzootic bovine leucosis.

Enzootic bovine leucosis. Enzootic bovine leucosis is associated with infection by bovine leucosis virus. The incubation period is measured in years and a minority of infected animals develop clinical signs. The disease is widespread in Europe and elsewhere and can cause significant economic loss. The epidemiology is incompletely understood and findings from one cattle production system may not be directly applicable to another. Major control programmes exist in Denmark and West Germany and control schemes are being developed elsewhere. Eradication of enzootic bovine leucosis has been established as a goal in the EEC and research is revealing the ways in which this goal may be attained. To be effective, control and epidemiological monitoring must be interactive. Recently introduced serological tests, of improved sensitivity, provide a valuable tool.


Interaction of a combined feline viral rhinotracheitis-feline calicivirus vaccine and the FVR carrier state. The effect of field feline viral rhinotracheitis (FVR) virus challenge on cats previously vaccinated with a combined FVR/feline calicivirus intramuscular vaccine was studied in relation to the development of an FVR carrier state. There was no virus shedding of either of the two vaccine viruses following vaccination. Treatment with corticosteroid 60 days after vaccination and before challenge with FVR virus did not induce virus re-excretion in vaccinates or controls; neither did similar treatment induce shedding 63 days after challenge of both vaccinates and controls with virulent field virus. After a further 55 days however, FVR virus shedding was elicited in one of four previously vaccinated and challenged cats compared with two of four unvaccinated and challenged controls. Two sentinel cats remained virologically and serologically free of FVR throughout. The vaccine was shown to be effective in controlling the disease; 12 weeks after initial vaccination no clinical signs were seen in three of four cats following intranasal challenge with 10(5)CCID50 of virulent field FVR virus, and a mild transient unilateral ocular and nasal discharge was seen in the remaining cat for one day only. Severe clinical signs of approximately 10 days' duration were seen in all four unvaccinated challenged controls. The virological and serological responses of the cats were also recorded.


[Serological diagnosis of bovine leukosis using the agar gel diffusion test]. Experiments were carried out to use the agar gel diffusion test (AGDT) in the study of bovine leukosis. An antigen was obtained from FLK-24 cells with a latent leukosis virus infection, which proved specific and suitable for AGDT. A total of 134 serum samples were examined applyig AGDT, for the presence of specific antibodies, originating from cattle of herds in which leukosis had been hematologically demonstrated. Investigated were also 56 sera of animals presenting an unknown clinical picture. Positive reaction was established in 70 per cent of the sera taken from herds with hematologically demonstrated leukosis. The specificity of AGDT was confirmed through establishing specific antibodies also in sera of lambs experimentally infected with a virus from a FBL-J 5 cell line with a latent infection. It is admitted that AGDT is a specific, sensitive, and readily applicable method, which can successfully be used in the diagnostics of leukosis in this country.


Malignant giant cell tumor of bone. Fine structure and localization of acid phosphatase. The fine structure of the different cell types constituting a primary malignant giant cell tumor of bone has been studied and the localization of acid phosphatase in relation to the subcellular organelles been demonstrated. Three distinct cell types with characteristic ultrastructural features were observed: giant cells, fibroblast-like cells, and cells with abundant lipid inclusions and mitochondria. Certain differences were noted between these three cell types and their counterparts in benign giant cell tumors of bone (described in a separate report). The enzyme histochemical and morphological data suggested that the giant cells in the malignant tumor might possess a more active and expansive lysosomal apparatus than corresponding cells in the benign variant.


Electron microscopic findings in four cases of nasopharyngeal fibroma. Light and electron microscopic investigations of four cases of juvenile nasopharyngeal fibroma revealed characteristic structures; a fibrous stroma, an inclination to hyalinisation and formation of scar like tissue, a lacunar thin walled vascular component, large numbers of mast cells and of fibroblasts. The tumor fibroblasts contained different nuclear bodies and particles. There existed five different types of more or less complex spherical bodies. The previously described nuclear electron dense particles with an electron lucent halo could be divided into four groups measuring 60, 90, 150, and 300 nm in diameter. In addition to the previously described ultrastructural properties of the tumors, the nuclei of the tumor fibroblasts were found to contain virus like particles. These particles were less electron dense, measuring 40 to 55 nm in diameter and arranged in groups throughout the nucleoplasm; they were different from chromatin condensations and from perichromatin granules. The structure and the size of the smallest particles was not comparable with the other nuclear inclusions.