Cyclic AMP modulates microvillus formation and agglutinability in transformed and normal mouse fibroblasts.

We have utilized dark field microscopy to observe the surface microstructure of living cultured cells. Using this method, we have found that dibutyryl cAMP treatment causes regression of the numerous, long cell surface microvilli present on L929 cells. Thirty minutes after removal of dibutyryl cAMP, microvilli reappear. An inhibitor of phosphodiesterase (methylisobutylxanthine) and a stimulator of adenylate cyclase (prostaglandin E1), both of which raise cAMP levels, cause regression of microvilli in 15 min. Untransformed 3T3 cells show very few microvilli when viewed still attached to their substratum or after removal with EDTA. Treatment of these cells with trypsin causes the formation of numerous microvilli on their surface. When clumps of cells agglutinated by concanavalin A are examined by thin section electron microscopy, the cells are seen to be held together by a "forest" of interdigitating microvilli and only rarely is there apposition of the areas of membrane between microvilli. At the same time the distribution of surface-bound concanavalin A was examined using immunofluorescent light microscopy, and concanavalin A was found to be uniformly distributed over the cell surface. We propose that agglutinability of mouse and rat fibroblasts is regulated through the modulation of cell surface microvilli by cAMP, and that transformed cells are highly agglutinable because their low cAMP levels result in the formation of numerous surface microvilli.


Physical characteristics of the gonadotropin receptor-hormone complexes formed in vivo and in vitro.

The physical properties of detergent-solubilized gonadotropin receptor-hormone complexes, determined by density gradient centrifugation and gel filtration, were compared after in vivo and in vitro labeling of specific ovarian binding sites with radioiodinated human chorionic gonadotropin (hCG). Following intravenous administration of biologically active 125I-labeled hCG, up to 50% of the gonadotropin tracer was bound to the luteinized ovaries of immature female rats treated with pregnant mare serum/human chorionic gonadotropin. Comparable binding of 125I-labeled hCG was observed after equilibration of ovarian particles with the labeled hormone in vitro. The sedimentation properties of the solubilized receptor-hormone complexes formed in vivo were identical with those derived for the corresponding complexes formed in vitro and extracted with Triton X-100 and Lubrol PX, with sedimentation constants of 8.8 S for the Triton-solubilized complex and 7.0 S for the complex extracted with Lubrol PX. During analytical gel filtration of the Triton-solubilized receptor-hormone complex on Sepharose 6B in 0.1% Triton X-100, the partition coefficient (Kav) of the "in vivo" complex (0.32) was not significantly different from that of the complex formed in vitro (0.29). Gel filtration of the Lubrol-solubilized ovarian particles on Sepharose 6B in 0.5% Lubrol PX gave Kav values for the "in vivo" and "in vitro" labeled complexes of 0.36 and 0.32, respectively. These findings demonstrate that the physical properties of size and shape which determine the partition coefficient and sedimentation characteristics of detergent-solubilized gonadotropin receptor-hormone complexes formed in vitro are not distinguishable from those of the complexes extracted after specific interaction of the ovarian gonadotropin receptors with radioiodinated hCG in vivo.


Regulation of herpesvirus macromolecular synthesis: sequential transition of polypeptide synthesis requires functional viral polypeptides.

It was previously shown that virus-specific polypeptides made in HEp-2 cells infected with herpes simplex 1 form three groups designated alpha, beta, and gamma whose synthesis is coordinately regulated and sequentially ordered. This report shows that one or more functional alpha polypeptides are necessary to turn on the synthesis of beta and gamma groups, and conversely, one or more polypeptides in the latter groups turn off the synthesis of alpha polypeptides. Specifically, infected cells maintained in medium containing either canavanine, an analogue of arginine, or azetidine-2-carboxylic acid an analogue of proline and hydroxyproline, synthesized alpha polypeptide at rates comparable to maximal rates in untreated infected cells but did not undergo the normal transition to beta and gamma polypeptide synthesis. The transition to gamma polypeptide synthesis and shut-off of synthesis of earlier polypeptide groups proceeded normally if addition of canavanine was delayed until at least 4-5 hr after infection. Addition of canavanine after the onset of beta and gamma polypeptide synthesis, i.e., between 2 and 3.5 hr after infection, resulted in sustained, simultaneous synthesis of all three polypeptide groups, a phenomenon not seen in untreated infected cells. Canavanine-treated infected cells, synthesizing alpha polypeptides, recovered the capacity to make beta and gamma polypeptides after removal of the analogue, but only after a 1-to 2-hr delay compared with infected untreated cells. The data indicate that the on and off controls inherent in the cascade regulation of viral polypeptide synthesis are mediated by one or more polypeptides in each group at transcriptional or post-transcriptional levels.


Tumor-inhibiting properties of the neutral P-O-ethyl ester of adenosine 3':5'-monophosphate in correlation with its crystal and molecular structure.

The P-O-ethyl ester of cAMP has been synthesized, its inhibition of solid and ascites tumors studied, and its pattern of urinary excretion followed. Et-cAMP is more effective than cAMP against solid sarcoma 180 in mice and against Ehrlich ascites carcinoma cells in tissue culture. The urinary excretion pattern of injected E-t-cAMP suggests that about two-thirds of the injected dose (13 mumol per animal) is retained in the rat rather than being promptly excreted. Liver slice studies of the effect on glycogenolysis suggest that the Et-cAMP is converted to cAMP intracellularly. The compound crystallizes in space group P21 with one molecule per asymmetric unit. The base ring has the anti conformation. The ethyl group is endo to the base ring and is axial in the flattened chair-conformer six-membered ring formed by the 3'-5' O-P-O cyclization. In most other respects the structure of the compound is closely similar to the known structures of other cyclic nucleotides.


Biochemical procedure for production of small deletions in simian virus 40 DNA.

A simple biochemical procedure for producing small deletions (15 to 50 base pairs) at virtually any location in simian virus 40 DNA has been developed. The steps involved are: cleavage of the closed-circular DNA to produce a linear structure followed by 5'-exonuclease digestion to expose a short single-stranded segment at each 3' end of the molecule. Mutants containing deletions at the site of the cleavage are obtained by infecting permissive monkey kidney cells with the exonuclease-treated DNA in the presence or absence of a helper DNA (depending upon whether or not the site of cleavage and therefore the deletion occurred in a gene required for vegetative multiplication). In this paper viable mutants with deletions at the HpaII endonuclease cleavage site (0.735 map position) and defective trans-complementable mutants with deletions at the EcoRI endonuclease cleavage site (0/1.0 map position) were isolated.


Somatic cell hybrids between mouse peritoneal macrophages and simian-virus-40-transformed human cells: II. Presence of human chromosome 7 carrying simin virus 40 genome in cells of tumors induced by hybrid cells.

Cells derived from tumors induced in "nude" mice after injection of cells that were hybrids between mouse peritoneal macrophages and simian virus 40 (SV40)-transformed human cells were found to retain the human chromosome 7 carrying the SV40 genome, and indicate that the presence of human chromosome 7 carrying the SV40 genome is responsible for the expression of the tumorigenic phenotype in the hybrid cells.


Estrogen-directed synthesis of specific prostaglandins in uterus.

Endogenous uterine prostaglandin levels were monitored in the cycling rat and prostaglandins of the F-type were found to rise at proestrus when estrogen levels have been shown to be maximal. Evidence that this is an estrogen-induced event is furnished by the finding that estradiol-17beta caused a similar rise in prostaglandin F levels in uteri of ovariectomized rats, an action blocked by coadministration of progesterone. Examination of in vitro prostaglandin synthetase by uterine microsomal fractions from cycling rats and estrogen-treated ovariectomized rats revealed that the action of estrogen to control prostaglandin synthesis in a directional manner is accomplished, at least in part, by regulation of the prostaglandin synthetase complex, resulting in a patterned alteration in the ratio of prostaglandin F to prostaglandin E. These results demonstrate that prostaglandins are involved in the expression of estrogen action in the rat uterus.


Macromolecular enzymatic product of NAD+ in liver mitochondria.

Rat liver mitochondria contain a Mg2+-requiring system that transfers the ADP-ribose moiety of NAD+ to an acceptor protein. The enzyme system was extracted in a soluble form and the ADP-ribosylated protein product was isolated by hydroxyapatite and Sephadex chromatography. The ADP-ribosylated protein product has a molecular weight of 100,000 and can be dissociated into subunits of 50,000 daltons by sodium dodecyl sulfate gel electrophoresis. Incubation of the isotopically labeled ADP-ribosylated protein with nicotinamide and a mitochondrial extract yields labeled NAD+, indicating apparent reversibility of the reaction. Enzymatic degradation of the ADP-ribosylated protein with snake venom phosphodiesterase liberates AMP and ADP-ribose or its isomer. Identification of these products and reversibility of the reaction show that the ADP-ribose moiety of NAD+ is the molecular species that is transferred to the acceptor protein. A fraction of the protein-bound ADP-ribose appears to be present as an an oligomer. The enzymatic protein-ADP-ribosylating reaction is inhibited by nicotinamide, ADP-ribose, the fluorophosphate of AMP, and picrylsulfonic acid.


Magnetic resonance spectra of membranes.

A number of electron and nuclear magnetic resonance studies of model membranes, and biological membranes, involve time-dependent magnetic interactions diffusion relative to one another. The two-dimensional character of this motion can have a special, large effect on magnetic resonance line shapes, and relaxation rates, because of the long-time tail of the correlation function for magnetic interactions modulated by this motion. Equations are given for the specific case in which nuclear relaxation rates are enhanced due to dipolar interactions with membrane-bound spin labels. An experimental study of spin-label-enhanced 13-C nuclear relaxation in unsonicated dispersions of phosphatidylcholine is accounted for with this theory, together with the previously reported lipid diffusion constant of D congruent to 2 times 10- minus 8 cm-2/sec. Our analysis of previously reported 1-H and 13-C nuclear relaxation rates in small phospholipid vesicles produced by sonication suggests that the rate of lateral diffusion in these small vesicles may be significantly larger than 10- minus 8 cm-2/sec.


Locus on human X chromosome for dihydrotestosterone receptor and androgen insensitivity.

We have studied androgen binding characteristics of cells for androgen insensitivity from an obligate heterozygote, who is the mother of three male pseudohermaphrodites with dihydrotestosterone (17beta-hydroxy-5alpha-androstan-3-one) receptor deficiency. Specific dihydrotestosterone binding by skin fibroblasts from her wrist and pubis is within the normal range and indicates that androgen insensitivity is X-linked in man and is therefore homologous to the tfm locus in the mouse. A significant population of clones from the heterozygote has deficient receptor activity, and this finding is compatible with inactivation of one X-linked allele at this locus. Our results indicate the presence of a gene on the human X chromosome that is essential for normal male sexual differentiation and provide further evidence for homology between X chromosomes of all mammals.


Epstein-Barr virus genomes with properties of circular DNA molecules in carrier cells.

A high-density fraction of high-molecular-weight DNA was isolated from the human lymphoid cell line Raji. This cell line contains 50 to 60 virus genome equivalents of Epstein-Barr virus DNA per cell, and the high-density DNA fraction was 10-fold enriched in such viral sequences. Sedimentation analysis on neutral glycerol gradients, followed by hybridization experiments with viral complementary RNA, showed that most of the intracellular viral DNA sequences in this material did not cosediment with the cellular DNA, but were recovered as two distinct species with sedimentation coefficients of 100 S and 65 S. These two forms sediment 1.70-1.75 and 1.10-1.12 times as fast as the linear Epstein-Barr virus DNA from virus particles, and thus have the hydrodynamic properties of a covalently closed circular form and a nicked (containing single-strand breaks) circular form of the virus genome. The 100S form also behaved as a covalently closed circular EBV DNA molecule on gradient centrifugation in CsC1/propidium diiodide, It would appear that latent Epstein-Barr virus DNA has the properties of a mammalian episome, and that both nonintegrated and integrated viral DNA sequences can be isolated from carrier cells.


Cell cycle specific fluctuations in adenosine 3':5'-cyclic monophosphate and polyamines of Chinese hamster cells.

Chinese hamster V79 cells were synchronized by mitotic selection, which resulted in approximately 95% synchrony. The adenosine 3':5'-cyclic monophosphate level was elevated within 3 hr (G1 phase) and reached a level 2-fold higher than in early G1 within 6 hr (early S phase). An increase in ornithine decarboxylase activity (6-ornithine carboxy-lyase, EC 4.1.1.17), the initial enzyme in the polyamine biosynthetic pathway, was detected within 4 hr and was maximal at 8 hr. Since about 20% of the cells were labeled with [3-H]thymidine at 4 hr, ornithine decarboxylase exhibits cell-cycle specific activity starting in late G1 and continuing through middle S phase. The activity of S-adenosylmethionine decarboxylase (S-adenosyl-L-methionine carboxylase, EC 4.1.1.50) increased within 5 hr, i.e., early S phase. It is suggested on the basis of these data and other studies discussed herein that the increase in ornithine decarboxylase activity, which parallels closely the elevation in cyclic AMP, is an example of adenosine 3':5'-cyclic monophosphate-mediated protein synthesis.


On the question of translocation of heart cAMP-dependent protein kinase.

Rat hearts were perfused with epinephrine and/or 1-methyl-3-isobutylxanthine for 2 min. These agents raised the concentration of cAMP and increased the fraction of cAMP-dependent protein kinase (EC 2.7.1.70) in the active form. However, the content of cAMP-dependent protein kinase in the soluble fraction of homogenates of these hearts was reduced and the amount in the particulate fraction was increased. A similar redistribution was obtained by adding cAMP to homogenates of control hearts. The reduction in soluble protein kinase content was due to apparent binding of the free catalytic subunit of the enzyme to particulate material (12,000 times g pellet) in media of low ionic strength (smaller than 100 mM KCl). The amount bound was, therefore, proportional to the dissociation of the holoenzyme. The binding was not altered by prior boiling or trypsin treatment of the particulate material, but it was prevented or reversed by the addition of 150 mM KCl. The catalytic subunit of the protein kinase from heart also bound to particulate fractions from liver or Escherichia coli and to various denatured proteins. These findings suggest that the protein kinase activity of membranes and particulate fractions has frequently been overestimated, since isolation of particulate materials has usually been carried out at low ionic strength. The data also imply that intracellular translocation of the protein kinase catalytic subunit, at least in heart tissue, is of questionable physiological significance.


RNA of replication-defective strains of Rous sarcoma virus.

The RNA of a replication-defective (rd) mutant, isolated from stocks of nondefective (nd) Schmidt-Ruppin Rous sarcoma virus of subgroup A (SR-A) and termed SR-N8, was compared to the RNAs of SR-A, of a transformation-defective derivative of SR-A (td SR-A) and of rd Bryan Rous sarcoma virus, RSV (minus). The molecular mass of the 30-40S species of SR-N8 RNA was estimated to be 21% (congruent to 7.5 to 8 times 10-5 daltons) smaller than that of SR-A by (i) electrophoresis in polyacrylamide gels and (ii) analyses of RNA complexity based on RNase T1-resistant oligonucleotides. ST-N8 shares probably all (=14) of its large RNase T1-resistant oligonucleotides with the RNA of SR-A as judged from the chromatographic distribution and the RNase A-resistant fragments obtained from RNase T1-resistant oligonucleotides. However, SR-N8 RNA lacked six large oligonucleotides which were present in the RNAs of SR-A and td SR-A. Conversely, the RNAs of SR-A, and of SR-N8 contained two oligonucleotides not found in td SR-A. The RNA of SR-N8 was found to differ from that of RSV (minus) in its electrophoretic mobility and its fingerprint pattern. It is concluded that the RNA of SR-N8 was generated by a deletion of SR-A. The extent of this deletion is compatible with the notion that the genetic information for the large viral envelope glycoprotein (molecular mass = 70,000-85,000 daltons) has been lost from the RNA of SR-A to yield SR-N8 RNA. From a comparison of td and rd deletion mutants, it appears that loss of different functions corresponds to the absence of different oligonucleotides in their RNA.


Induction of growth in resting fibroblastic cell cultures by Ca++.

Of all the components of the culture medium, only CaCl2 induces DNA replication when added to resting cultures of Balb/c 3T3 cells. The effect is present even in a serum-free medium. Increasing the Ca++ concentration above the standard 1.8 mM in the medium of a new culture increases the total number of cells ultimately produced, without affecting the initial cell growth rate. This effect is synergistic with that of serum. The elevated Ca++ concentration also induces striking morphological changes. The Ca++ effect could not be reproduced by a Ca++ ionophore. These observations afford a new tool for studying how the various intracellular events following the addition of growth factors to resting cultures are involved in the control of cellular growth.


Nuclear magnetic resonance spectroscopy: reinvestigation of carbon-13 spin-lattice relaxation time measurements of amino acids.

The carbon-13 spin-lattice relaxation times (T1) of glycine have been measured as a function of pD and concentration. Contrary to previously reported findings, no significant dependence was observed on either pD or concentration. In addition, the T1 values reported here are much longer than those published earlier. The discrepancies arise from the presence of paramagnetic impurities in the earlier samples. For the carboxyl carbon, dipole-dipole relaxation is dominant in both D2O and H2O solution, and in H2O there is a significant intermolecular dipolar contribution. Proton and oxygen relaxation times have also been measured. These, along with the carbon relaxation data, allow a discussion of the dynamics of glycine in solution.


T antigen binds to simian virus 40 DNA at the origin of replication.

A technique employing ferritin-conjugated antibody has been developed to visualize specific protein-DNA complexes in the electron microscope and has been used to demonstrate the preferential binding of simian virus 40 (SV40) T antigen at or near the origin of replication of SV40 DNA, 0.67 fractional length clockwise from the EcoRI restriction endonuclease cleavage site. urified covalently closed supercoiled circles of SV40DNA were treated with partially purified T antigen and the complex was stabilized by crosslinking with glutaraldehyde. Hamster antiT antigen gamma-globulin, ferritin-labeled goat anti-hamster gamma-globulin, and glutaraldehyde were then added sequentially. The location of the bound ferritin cores was measured with respect to the EcoRI cleavage site and the orientation of the cores relative to the ends of the DNA was determined with respect to the locations of Escherichia coli DNA unwinding protein, which binds to covalently closed supercoiled SV40 DNA at either of two preferred sites, 0.46 or 0.90 fractional length clockwide from the EcoRI cleavage site.


Cytotoxic effector cells specific for B Cell lines transformed by Epstein-Barr virus are present in patients with infectious mononucleosis.

Peripheral lymphoid cells, from 12 cases of acute infectious mononucleosis (IM), were tested in a micro chromium-51 release assay for cytotoxic activity against a variety of cell lines that did or did not carry the Epstein-Barr virus (EBV) genome. Unfractionated lymphocytes from these patients were cytotoxic to both types of cell lines, as were lymphocytes from healthy individuals. If, however, lymphocytes bearing complement receptors were removed, the residual IM lymphocyte fraction was specifically cytotoxic for EBV-genome-carrying cell lines. The residual lymphocyte fraction in normal donors had no such effect. Heterophile-positive IM is caused by EBV, and these results indicate that, during the acute phase of this disease, patients harbor killer cells, probably T cells, which specifically kill EBV-genome-carrying B cells in vitro. No such specificity for EBV-genome-psitive target cells was found in normal lymphocytes stimulated in vitro with autologous EBV-genome-positive lymphoblastoid cells. Such stimulated cells were highly cytotoxic to both genome-positive and negative lines after removal of complement receptor-positive lymphocytes.


Functional intermediates in reaction of cytochrome oxidase with oxygen.

The development of a low temperature kinetic method for the flash photolysis of the compounds of membrane-bound cytochrome a3 with carbon monoxide in the presence of oxygen affords evidence for three categories of functional intermediate compounds of cytochrome a3 and oxygen. The three classes are identified as follows: Compounds of Type A are considered to be "oxy" compounds of the ferrous heme. They have the composition a3-2+. O2. Compounds of Type B are considered to be peroxide compounds (CU-2+A3-3+ O-2= or CU-2+A3-3+ O2H2) or the equivalent heme Fe-Cu peroxide bridge structures. Compounds of Type C are formed from the ferricyanide pretreated oxidase and may involve higher oxidation states of the heme iron such as quadrivalent iron, and peroxide. Kinetic and equilibrium studies show these compounds to be functional in oxygen reduction in the sequence A yields B yield cytochromes a, c, c1, etc.


Biologic and antigenic similarity of virus-induced migration inhibition factor to conventional, lymphocyte-derived migration inhibition factor.

Macrophage migration inhibition factor (MIF) is one of a class of lymphocyte-derived mediator substances (lymphokines) which plays a role in the mechanism of cellular immunity. A variety of other soluble factors produced by non-lymphoid cells have been shown to have effects on macrophage mobility similar to that of MIF. In the present study we demonstrate that one such factor, (MIFV) derived from simian virus 40-infected kidney cells in culture, has several other properties in common with lymphocyte-derived MIF (MIFL), MIFV can be  adsorbed on Sepharose bead columns conjugated with an antiserum prepared against MIFL, demonstrating at least some antigenic similarity. Moreover, MIFV can substitute for MIFL in an in vivo system involving the suppression of cutaneous manifestations of cellular immunity by intravenous injection of the lymphokine. These observations, taken in conjunction with the similarity of the in vitro effect of MIFV and MIFL, and their similar chromatographic behavior, suggest that MIFV and MIFL may be identical molecular species.


Adrenal Cortical Ribonuclease H (hybrid) (38494).

Ribonuclease Hybrid (RNaseH) from adrenal cortical tissue has been characterized. RNase H specifically degrades the RNA strand of purified RNA:DNA hybrids but is inactive on single- or double-stranded RNA or on DNA. The mode of clevage by RNase H is endonucleolytic, producing oligoribonucleoties with 3'-hydroxyl and 5'- phosphate termini. ACTH administration to guinea pigs results in no detectable change in adrenal cortical RNase H activity at times when changes in DNA synthesis are marked.


Elevated cyclic AMP levels in mouse lymphoid tissue after stimulation by cholera enterotoxin in vitro (38468)

Addition of CT to suspensions of thymus, lymph node, spleen, or bone marrow cells in vitro resulted in a marked accumulation of cAMP with peak levels occurring 4-5 hr after incubation  of cells with CT. Thymus cells showed the largest increase in cAMP, approximately 40-fold at 10  ng/ml CT. Bone marrow cells accumulated the least cAMP (1.5x), while intermediate levels were observed for spleen and lymph node cells (10-12x). Antiserum to CT prevented stimulation of increased cAMP levels. Repopulation studies using X-irradiated mice also showed that thymus-derived spleen cells accumulated more cAMP/10-7 cells than spleen cells from recipients given spleen or marrow cells. Spleen cells from athymic (nu/nu) mice also responded much less than did spleen cells from normal mice. Thymocytes appeared to bind CT to a greater degree than bone marrow cells. Spleen and lymph node cell suspensions also contained CT-binding  cells and the number of CT-binding cells in these peripheral lymphoid tissues appeared approximately equal to the summation of the numbers observed in thymocyte and bone marrow cell suspensions. Stimulation of cAMP in lymphoid cells, especially thymocytes, by CT provides a pharmacological tool to investigate the mechanism and role of this nucleotide in the early events of antibody formation.


Isolation of a new enterovirus (38506).

During 1972 an apparently new enterovirus with characteristics of the Coxsackievirus group A was isolated from 11 residents of New York State, 10 of whom had central nervous system disease. The representative strain S/Albany 1/72 has the physical and chemical properties of an enterovirus and causes the pathology typical for Coxsackievirus group A in 1-day-old mice. Cross-neutralization tests indicated that the virus is distinct from currently recognized enteric viruses of man. Antibody levels to the representative strain were moderate to high in patients and were low to moderate in 26% of healthy individuals tested.


Preparation of rhinovirus antisera in the South American hystricomorph rodent, Octodon degus (38507).

Rhinovirus antisera have been prepared for rhinoviruses (RV) 7,9,26,32,67 and 87 in guinea pigs and in degus. Titers achieved were either similar in the 2 animals (RV7) somewhat higher in the degu (RV9 and RV32) or clearly higher in the degu (RV26, RV67 and RV87). Specificity of the antisera was similar in both animals. In special instances where it is difficult to prepare high-titered rhinovirus antisera in the guinea pig, the degu offers an attractive alternative source.


Stimulation of macromolecular synethesis in guinea pig cells by human CMV.

Nonproductive human CMV  infection of G.P. embryo cells induced cellular RNA and DNA synthesis. RNA species synthesized in abortively infected G.P. cells included ribosomal 28S, 18S, and 4S transfer RNA. Virus inactivated with heat and UV light failed to induce cellular RNA and DNA synthesis. Evidence for increased DNA and RNA synthesis in infected cells was provided by studies of:(a) incorporation of thymidine and uridine, (b) extraction  and separation of cellular DNA and RNA on sedimentation gradients, (c) autoradiography, and (d) measurement of mitotic index.


In vitro interaction of estrogen and prolactin on hormone-dependent rat mammory tumors (38511).

Hormone-dependent, regressing mammary tumors in rats were incubated in a chemically defined culture medium containing tritium labeled leucine at 37 degrees for 2 hr. The rate of tritium incorporation into the protein fraction by these tissues was measured with and without the addition of 17beta-estradiol and/or ovine prolactin into the medium. Tumor tissue responded to either estrogen or porlactin by a 6-7% increase in the rate of 3-H-leucine incorporation. When the incubation medium contained both the  hormones, the increase in the rate of 3-H leucine incorporation was 19% which was significantly greater than the sum of the effect of each single hormone measured independelntly. These results demonstrate a synergistic effect between estrogen and prolactin on rat mammary tumors.


Glucose tolerance and pancreatic ultrastructure in mice with long-term diabetes induced by EMC virus (M variant) (38518).

Of 27 young male CD-1 mice infected with the M variant of the encephalomycarditis (EMC) virus and surviving for 11 or 12 mo, all but three had normal glucose tolerance curves, and all displayed normal islet cell morphology, at the time of sacrifice, in spite of an initital hyperglycemic response. Three animals maintained diabetic glucose tolerance curves one year after infection but they, too, failed to show significant morphological alterations in their pancreatic beta cells.


Photodynamic inactivation of herpesvirus hominins by methylene blue (38524).

Methylene blue, in a concentraion of 10-5M was virostatic in the presence of light but not in the dark for 31 of 37 strains of fresh isolates of herpesvirus hominis. Resistance to the dye developed during treatment. This photodynamic pattern was almost identical to that of neutral red which produced an identical effect. This was not ture for proflavine which was active in the dark in many instances. Cross insensitivity between proflavine and methylene blue was not observed.


Comparison of the lipopretein profiles and the effect of N-phenylpropyl-N-benzyloxy acetamide in primates (38473).

Analytical ultracentrifugation showed Cebus and Rhesus monkeys had two low density components while only one was present in Squirrel monkeys. In untreated or W1372 treated monkeys, neither chylomicrons nor very low density lipoproteins were detected on analytical ultracentrifugation. Chylomicrons were not observed on agarose gel electrophoresis. Ultracentrifugal analysis showed W1372 treatment decreased the amount of LDL in all animals and also the HDL in Cebus monkeys on an atherogenic diet. Both untreated and W1372 treated Cebus monkeys on an atherogenic diet had abnormal amounts of LDL and HDL, while the LDL in treated animals occurred as multiple peaks. This was also evident on agarose gel  electrophoresis. Accumulation of lipds in the liver and decrease of serum lipids indicated W1372 prevented release of lipoproteins from the liver.


The effect of various prostaglandins and a prostaglandin synthetase inhibitor on rat anterior pituitary cyclic AMP levels and hormone release in vitro (38475).

(U)Prostaglandins E-1, E2,F-1alpha or F-2 alpha significantly increased the release of GH, with a parallel increase in intracellular cAMP concentrations, while they only protentiated HE-stimulated TSH release. (2) None of the prostaglandins examined consistently effected either the basal or HE-altered release of LH,FSH or prolactin. (3) The prostaglandin synthetase inhibitor, indomethacin, inhibited GH and TSH release and, at high doses of the drug, inhibited prolactin release. In contrast, the drug appeared to potentiate both He and sLRF-stimulated gonadotropin release. It had no significant effect on intracellular cAMP concentration.


An improved method of fixation for immunofluorescent detection of SV40 T-antigen in infected human fibroblasts.

Improved fixation for immunofluorescence staining in the SV40 T-antigen test has been developed utilizing ethanol in place of acetone.  The modified method improves resolution, results in smaller variability of cell numbers, and reduces the time of assay, while maintaining its sensitivity in discriminating between normal and cancer prone groups.


Stimulatory effects of peptides on growth of the free-living nematode Caenorhabditis briggsae.

Evidence is presented that peptide products of hydrolysis of casein, including some di- and tripeptides, but not the  constituent amino acids, can stimulate growth of C. briggsae in defined basal medium supplemented with cytochrome C and B-sitosterol.  Peptide activity may raise from amino acid imbalances in the medium which causes competitive inhibition of uptake of essential amino acids.  Such activity precludes facilitation of heme transport as the sole function of growth factor in C. briggsae.  This is the first report in a nutritional role of peptides in invertebrate metazoa.


Effect of clofibrate on lipid metabolism in streptozotocin diabetic rats.

The effect of clofibrate (CPIB) on lipid metabolism was studied in male rats rendered diabetic by intravenous injection of 80 mg/kg of streptozotocin. After 1 wk, the rats received by gastric intubation 242 mg/kg/day of CPIB for 7 days. Liver lipid concentration remained unchanged in experimental diabetes and after treatment with CPIB; however, due to decreased liver weight, total liver lipids were lower in diabetic rats. Elevation of cholesterol, phospholipids, and triglycerides in the serum of diabetic rats was reversed by CPIB treatment. Hepatic cholesterol synthesis in diabetic rats was suppressed to approximately 1/10 of that in normal rats. Treatment with CPIB abolished this residual cholesterogenic activity. Diabetes had no effect on intestinal cholesterol synthesis; a slight increase was noted after CPIB treatment. Basal and norepinephrine-induced lipolysis in fat pads was elevated in diabetic rats; CPIB had no effect on these changes. The data show that the elevated serum lipids in diabetic rats are lowered by treatment with C-IB. It was concluded that the hypocholesterolemic activity of clofibrate in rats is not caused by its suppression of hepatic cholesterol synthesis.


Effects of "stress" on pulsatile luteinizing hormone release in ovariectomized rats.

The effects ov various "stresses" on pulsatile LH release in ovariectomized rats were investigated. Blood was withdrawn through atrial cannulas and replaced with saline at 10 min intervals for 190 min. Plasma LH concentration was suppressed in rats subjected to 30 or 60 min of immobilization during the collection period. On the contrary, sham ovariectomy 4 hr prior to blood collection, leg break or iv injection of ACTH or corticosterone during the collection period did not alter pulsatile patterns in LH release. The results indicate that LH release mechanisms are highly resistant to "stresses" but that immobilization can suppress LH release by an unknown mechanism.


Effects of endurance exercise on serum enzyme activities in the dog, pig and man.

Daily levels of GOT, GPT, AK and LDH did not change consistently with exercise in any species. The level of CPK each day before exercise presented a consistent rise during the week of exercise in the dog and man but not in the pig. By the third day of resting, all enzyme activities were at control levels. The probable tissue of origin for AK and LDH at rest as well as after exercise was the heart, liver, skeletal muscle and kidney in all three species. Serum CPK was the most sensitive index of acute exercise stress in the trained dogs and men. We have shown that the pig, which exhibits responses to exercise resembling those of man, can be exercise trained. The pig may, therefore, be the more appropriate model for endurance exercise studies.


The effect of mycoplasmas on replication and plaquing ability of Herpes simplex virus.

M. arginini, an arginine utilizer, can decrease the yield of Herpes simplex virus, type 1 grown in Vero cells. M. arginini can also cause a reduction in number and size of plaques produced by HSV.  The reduction in titer and plaque size produced in M. arginini-infected cells can be reversed by supplementing medium with additional arginine. A. laidlawii, a nonarginine utilizing mycoplasma, had no effect on the growth of HSV.


Malignant transformation of hamster cells following infection with bovine herpesvirus (infectious bovine rhinotracheitis virus.

Hamster embryo cells, following infection with IBR virus, showed malignant transformation.  Hamsters of all ages, inbred or random bred, inoculated with two of the transformed cell lines developed solid tumors.  Preliminary characterization of the tumors induced by one of the cell lines has indicated undifferentiated sarcomas.  Viral specific antigen was detected in about 5% of the transformed cells and 10% of primary tumor cells in culture.  Viral specific antibody was detected in the serum of tumor-bearing hamsters by the indirect immunofluorescent method, but no neutralizing antibodies were found.  Infectious virus has not been recovered from either the transformed or tumor cells by cocultivation with bovine embryonic kidney cells.


Tumor antigens in hamsters with sarcomas associated with herpesvirus type 2.

Tumor-associated antigens (TTA) were demonstrated in preparations of hamster sarcomas associated with Herpes simplex virus type 2, as well as in the sera of tumor-bearing hamsters. An immunoadsorption-in-gel method was employed to demonstrate and purify the TAA  and anti-TAA. These results suggest the potential use of this technique for the demonstration of TAA or of anti-TAA in humans or animals with cancer.


Neurohypophyseal hormones and analogues: magnesium dependence and contraction of arterial smooth muscle.

The present quantitative results, using isolated rat aorta, demonstrate that different [Mg2+]o (i.e. 0.2, 1.2 and 6.0 mM) potentiate the contractile actions of a variety of neuohypophyseal hormones and synthetic analogues on vascular smooth muscle. [Mg2+]o can alter both the hormone-receptor affinities (H-RA) and intrinsic (contractile) activities (i.a.) of these peptides on vascular muscle; 1.2 mM [Mg2+]o (approximately that found in rat plasma) appears to optimize H-RA and i.a. on rat aortic smooth muscle. The presence of [Mg2+]o not only steepens the concentration-effect curves to the neurohypophyseal peptides but increases the maximum contractile responses as well. The present findings question that [Mg+]o potentiates responses to neurohypophyseal peptides by  vascular muscle solely by affecting H-RA. The present study supports the notion that Mg2+ potentiates responses to these peptides by acting at sites other than the receptor in mammalian vascular muscle. In addition, the present experiments suggest that the [Mg2+]o dependence of neurohypophyseal peptides on at lesast one mammalian vascular muscle-rat aorta- is directly rather than inversely proportional to the rat pressor potency of the molecules. Further, the vasopressin receptor which subserves contraction in mammalian blood vessels may differ in this respect from that in uterine smooth muscle.


Enhancement of interferon production in vitro: a property of tilorone-poly rL:rC/DEAE-Dextran.

Tilorone and Poly rI:rC, in the presence of DEAE-dextran, were found to exhibit a marked synergism with respect to the induction of interferon in L929 and primary mouse embryo fibroblasts, but not human foreskin fibroblasts, in cell cultures. The degree of synergism was proportional to the concentrations of tilorone and Poly rI:rC and was influenced by the times of addition of the compounds relative to each other.


The inhibitory effects of MgCl2 on the inactivation kinetics of poliovirus by urea.

By analyzing the inhibitory effects of Mg-2+ on the three-stage urea inactivation curve of poliovirus, it was concluded that urea inactivates poliovirus via a two-step reaction as follows: urea initially converts the native virus into an intermediate state which is still infectious but is now highly sensitive to inactivation. This reaction is unaffected by Mg-2+ and is reversible. In the subsequent reaction, the sensitized virus is either irreversibly inactivated by urea or reversibly stablized by Mg-2+. In addition, the inactivation curve revealed that a fraction of relatively stable virus population was established in the presence of urea and that the size of this persistent virus population depended on the concentration of urea. It was not determined whether urea induced the formation of this stable fraction of virus or merely selected for a preexisting stable population of virus. However, evidence was presented that (1) the persistent virus population was not due to the depletion or inactivation of urea in the sample, (2) whatever stabilized the virus, it could not be removed or reversed by simple dilution as in the case of Mg-2+, (3) no excess stabilizing material was made to stabilize the addition of untreated virus and finally (4) the persistent virus population was resistant to that concentration of urea in which it was observed but could be further inactivated by a higher concentration of urea, only to result in a smaller fraction of persistent virus population.


Evidence for conformational states of poliovirions: effects of cations on reactivity of poliovirions to guanidine.

The kinetics of inactivation of poliovirions were determined in the presence of various concentrations of monovalent and divalent ions, methionine, guanidine and combinations of these at various temperatures. Four states of the virion, under control of the ionic environment, could be recognized which were reversibly interchangeable and yet retained infectious potential. These states were detected by differential sensitivity of the virion to inactivation of infectivity by heat, urea and guanidine. The data are interpreted in terms of conformational structural changes under control of the ionic environment. It is proposed that these various structural conformations are meaningful to the various biologic functional states of the virus.


Effect of ascorbic acid on rhinovirus replication in WI-38 cells.

Exposure of WI-38 cells to ascorbic acid plus glutathione mixtures in growth medium for 2 days prior to infection with rhinovirus serotype 20 and during virus replication suppressed multicyclic but not single-cycle growth of the virus. Multicyclic growth was suppressed over the range of multiplicity of infection (m.o.i.) of 4 times 10-minus 1 minus 4 times 10-minus 4 PFU/cell. There was some suggestion that, in the presence of ascorbic acid plus glutathione, interferon was produced at the highest m.o.i. tested but at barely detectable levels.


Resticted growth of human cytomegalovirus in UV-irradiated WI-38 human fibroblasts.

The growth of human CMV was inhibited by uv irradiation of cells prior to infection or during the 48-hr latent period of virus replication but not after virus synthesis began. The duration of uv exposure sufficient to inhibit CMV replication was insufficient to inhibit replication of Herpes simplex and did not prevent uninfected cells from dividing normally. The effect of uv irradiation on CMV replication may have been mediated through prevention of the virus on host cell RNA(s) synthesis.


Adipose tissue glycerokinase activity in genetic and acquired obesity in rats and mice.

Glycerokinase activity in isolated fat cells was elevated in both Ob/Ob and Db/Db mice in comparison to their lean controls and this elevation was associated with obesity, hyperinsulinemia and hyperglycemia. In the other forms of acquired and genetic obesity in the rats and mice studied (also associated with hyperinsulinemia), adipose tissue glycerokinase activity was not elevated in comparison to lean control groups when expressed on a mg protein basis. It is concluded that the elevated glycerokinase activity is not due to the specific Db or Ob mutation, but is secondary to the obesity and hyperinsulinemia interacting with the similar genetic background in the  C57BL/KsJ and the C57BL/6J mouse strains.


Differentiation of nephrotensin from the renin angiotensin system.

An investigation of the relationship between nephrotensin and the renin angiotensin system was carred out. Nephrotensin was found in the plasma of rats with renal clip hypertension and with chemically induced kidney damage. There was no demonstrable correlation between presence of nephrotensin and plasma renin activity, and the pressor activity of nephrotensin was not altered by previous immunization of test animals with angiotensin II nor by pretreatment with angiotensin I converting enzyme inhibitor. These results indicate that nephrotensin is different from the components of the renin-angiotensin system.


Modification of surface membrane antigens by trypsin.

Cultured 3T3 and PY-3T3 mouse fibroblasts were subjected to mild treatment with twice crystallized trypsin and their immunogenicity tested in ICR/CD-1 mice. Surface antigenic modifications were observed using the foot pad swelling assay for detection of delayed hypersensitive responses. Mice immunized with trypsin-treated 3T3 cells showed cell-mediated immune reactions following challenge with either homologous antigen or untreated PY-3T3 fibroblasts. In contrast, treatment of the virus-transformed cells with trypsin decreased their antigenicity as determined by this in vivo assay.


A specific complement-fixation test for human hepatitis a employing CR326 virus antigen. Diagnosis and epidemiology.

A specific diagnostic complement-fixation test for hepatitis A antibody in human serum was described employing livers of marmosets infected with CR326 strain human hepatitis A virus. Persons with hepatitis A, but not hepatitis B, developed hepatitis A CF antibody shortly after the onset of illness and this persisted thereafter. Good agreement was noted in the development of CF and neutralizing antibodies in hepatitis A cases. Hepatitis A was shown to occur in a person with hepatitis B antigenemia and hepatitis B occurred in persons with hepatitis A antibody. Most persons with hepatitis A who were tested, but none of those with hepatitis B, developed increased anticomplementary activity in their sera at the time of onset of illness. At least one patient with hepatitis A developed antibody against normal liver that persisted. The possible inplications of this in relation to pathogenesis and to non-specific diagnostic tests in hepatitis were discussed. A limited epidemiologic study of a family outbreak of hepatitis in Costa Rica and of a group of young adults in our epidemic country acquire their infections at an early age and are immune thereafter; persons in areas of relatively low incidence may proceed into adulthood without experience with hepatitis A. The CF test should provide an excellent tool for diagnosis and for epidemiologic investigation of hepatitis A and should be of considerable value to detect hepatitis A virus in attempts to propagate the virus in cell culture.


Protein and RNA synthesis in follicles isolated from rabbit ovaries.

The in vitro synthesis of protein and RNA in follicles from mated and unmated rabbits has been studied by measuring precursor incorporation. Radioactive methionine incorporation into follicular proteins showed a transitory rise following coitus but fell to precoital levels in ovulated follicles. LH, FSH, and cyclic AMP all exerted an acute stimulatory action on the incorporation of amino acids into proteins by follicles from unmated rabbits. Radioactive uridine incorporation into follicular RNA showed the same temporal pattern of postcoital rise followed by a decline in incorporation rates in the period around ovulation. In acute incubations, FSH and cyclic AMP stimulated RNA synthesis; LH was without acute action on this process. The results of these studies suggest that a wave of follicular protein and RNA  synthesis precedes ovulation and may be partly related to the preovulatory surge in gonadotropins.


CD spectra and redox reactions of superoxide dismutase from Escherichia coli B: evidence for a Mn(III) enzyme.

Analysis of the metal content, CD and EPR spectra, and reaction with H(2)O(2) and ferrocyanide have been carried out on E. coli superoxide dismutase, which contains manganese as the metal prosthetic group. The results obtained indicate that the metal is present in the enzyme as a high spin Mn(III) complex of highly distorted octahedral symmetry, in the ratio of approximately 1 atom of manganese per enzyme dimer.


Evidence for a lesion in carbohydrate metabolism in fatty liver and kidney syndrome in chicks.

Liver and kidney slices from chicks affected with fatty liver and kidney syndrome (FLKS) were incubated in vitro with a variety of non-carbohydrate precursors and their ability to form glucose was studied. The results show that the affected liver was unable to form significant quantities of glucose from the precursors (gluconeogenesis). Glycogen breakdown was also drastically reduced because the tissue was almost devoid of this carbohydrate store. Blood chemistry revealed no evidence of overall liver malfunction but reflected the consequences of lack of gluconeogenesis. In contrast, kidney gluconeogenic activity was significantly higher than in the controls, suggesting an attempt by this organ to offset the reduced hepatic capability. Attempts to restore activity in vitro were made by adding known cofactors of gluconeogenesis. Asmall but significant improvement resulted from addition of biotin to liver slices.


Serum alkaline phosphatase activity in relation to liveweight of lambs.

Groups of lambs were fed four levels of a diet based upon oaten grain. Two groups gained weight rapidly, one less rapidly and the other lost liveweight over a 32-day period. Serum alkaline phosphatase activity was found to be closely correlated to liveweight, in a positive linear manner, in those lambs gaining weight rapidly. This relationship was similar in lambs of R or r blood group. Supplementation of the diet with limestone and or sodium chloride did not influence the relationship. This relationship is considered to reflect the influence of the dietary treatments upon the rate of skeletal metabolism.


A comparison in calves of the antigenicity of three strains of bovid herpesvirus 2.

Three strains of bovid herpesvirus 2, viz. Allerton, bovine mammillitis and 69/1LO were used to infect calves intradermally. Twenty-eight days later the immunity of the calves was challenged by intravenous injection of a homologous or heterologous strain. Challenge control calves developed a fever (greater than 40 degrees C) lasting several days and widespread skin lesions which varied with the strain. Homologous challenge of the primary infection produced neither skin lesions nor febrile response, except in one calf in which fever was noted on one day. Heterologous challenge did not cause skin lesions but fever occurred in 8/12 calves. In particular Allerton virus failed to protect completely against heterologous challenge. Despite minor differences evident in these experiments, it is recommended that these isolates should be considered as strains of the same virus--bovid herpesvirus 2.


The incidence of antibody to infectious  bovine rhinotracheitis virus in Kenyan cattle.

A total of 3204 cattle sera, collected between 1966 and 1974, from all seven provinces and 34 of the 42 districts of Kenya were screened in a neutralisation test for antibody to infectious bovine rhinotracheitis virus (IBRV). Antibody to IBRV was found in some sera from all the districts, with cattle over two years old showing the highest incidence. A small number of goat sera also showed some antibody. From the results obtained it is concluded that IBRV infection is widespread in Kenyan cattle.


Comparison of three methods for sampling the bovine upper respiratory tract for viruses.

A modified naso-pharyngeal swab with nylon bristles had certain advantages for sampling the bovine upper respiratory tract for viruses. The titre of parainfluenza virus type 3 recovered was greatest when this type of swab was used compared with two other methods of sampling.


[Diagnostic value of bone scintigraphy with technetium pyrophosphate. Study of 250 patients].

The authors report the results they obtained by bone scintigraphy using technetium pyrophosphate. In a study of 142 patients with cancer, the authors show, as others have done, that bone scintigraphy makes it possible to find bone metastases that are radiologically undetectable and they emphasize the importance of this discovery. In 7 patients with spondylodiscitis, of whom 1 was without radiological signs at the time the scintigraphy was carried out, the authors always observed localized vertebral hyperfixation and they noted that this examination can be valuable for distinguishing spondylodiscitis from pseudo-Pott's discarthroses and from the lesions of vertebral epiphysitis, which in their experience do not result in isotopic hyperfixation. In 7 patients with epiphyseal osteonecrosis, the authors observed isotopic hyperfixation before the appearance of radiological signs. In 12 patients with osteoporosis, the authors observed hyperfixation in bone in certain compressed vertebrae, whereas other vertebrae that had probably been compressed some considerable time earlier did not fix the isotope excessively. They never observed hyperfixation in vertebrae that were not compressed. Among 5 patients with ankylosing spondylitis with radiological signs of sacro-iliac arthritis, the authors observed sacro-iliac hyperfixation in only 3 cases. Two other patients who had signs indicating ankylosing spondylarthritis, but were without radiological signs of sacro-iliac arthritis did not show sacro-iliac hyperfixation of the isotope. Among 7 patients with Paget's disease, the authors observed hyperfixation in all the bones with radiological signs of disease; in addition, in 3 patients, there was also hyperfixation in certain bones that were radiologically clear.


Stress polycythemia--a pathophysiologic study.

Two patients with "Gaisböck syndrome" (stress polycythemia) were found to have been under chronic "stress" with REM sleep decrease, decreased nocturnal ADH-like activity and consequent nocturnal water loss. The role of REM decrease with chronic "stress" in causing secondary decrease in nocturnal ADH-like activity was emphasized as the means by which "stress" caused the blood volume changes seen with these two patients. Some factors that possibly could play a role in the pathophysiology of this syndrome were ruled out and other factors not studied here were considered for future investigation.


The prevention of iron dificency anemia in infants.

The prevention of iron deficiency requires attention to the toal feeding regimen as well as to its iron content. The use of fresh cow's milk in early infancy, a common practice, is associated with the development of measurable degrees of iron deficiency. In many instances excessive amounts of milk produce severe iron deficiency anemia during the second year of life in infants of greater than average birth weight. The psychological factors leading to such milk intakes have been reviewed. Present evidence supports the hypothesis that occult blood loss is a major etiologic factor in iron deficiency in infancy. Although the mechaisms of intestinal damage by fresh cow's milk protein is highly suspect and that the concentration of protein in the formula may play a significant role. What about iron fortification of infant formulas? Itis becoming clear that fullterm infants fed prepared formulas having protein concentrations approaching that of human milk and fed diets containing about 7 mg of iron daily after the age of 3 months infrequently develop biochemical iron deficiency. The addition of iron to the formula at levels of 10 to 12 mg/gt is assurance that their intake is adequate, whatever the iron intake from other foods. Low-birth-weight infants have increased iron needs because of their rapid growth. Iron-ofrtified formulas are necessary for these infants. Since iron from fortified formulas is well absorbed during the first three months of life, even if it is not immediately used for hemoglobin formation, an inccrease in the iron stores will occur...


Glycogen content in the adrenals and liver of rat embryos depending on the level of adrenocorticotropic functions.

The functional state of the adrenals of rats during prenatal development was studied. The accumulation of glycogen in the adrenals and liver in 15.5-21.5-day old embryos was used as the index. The maximum glycogen concentration in the adrenals and the minimum in the liver were detected in 15.5-16.5-day embryos, i.e., before the adrenals had begun to produce glucocorticoids. A sharp decrease in glycogen in the adrenals was observed in 17.5-day embryos, and an appreciable accumulation in the liver in 18.5-day embryos, which coincides with the beginning of the glucocorticoid function. Decapitation and encephalectomy performed at an age of 17.5-19.5 days of development of the embryos, were invariably accompanied by a significant increase in the glycogen content in the adrenals and a decrease in its level in the liver in the case of investigation after two days. This indicates an active participation of the hypothalamo-pituitary system in the regulation of the adrenocortical function in the late days of embryonic development.


The in vivo uptake of tritiated estradiol in carcinoma of the breast.

In human beings, the uptake of radiolabeled estradiol suppressed by carcinomatous breast tissue in vivo in relation to adipose breast tissue has been studied in 20 patients. The ratio of tumor uptake to fat tissue uptake was correlated with the age of the patient, menopausal status, presence of axillary metastases at time of mastectomy, and the response to endocrine therapy if metastases were presented. There was no correlation between a higher uptake of estradiol by the tumor and age of the patient, menopausal status, or presence of axillary metastases at time of mastectomy. Patients with a higher uptake of estradiol tended to respond favorably to endocrine treatment.


Glandular tumors of the palate.

Salivary gland tumors of the palate originate in the minor salivary glands located in the glandular zone of the palate. Approximately 7 per cent of all tumors of the salivary gland occur in the palate, and approximately one-half of such tumors are cancers. The salivary gland tumor is more often found in the hard palate. Of the 90 salivary gland tumors of the palate reported herein, 49 per cent were benign, and all were pleomorphic adenomas. Of the cancers, 70 per cent were cylindromas, 15 per cent mucoepidermoid cancers, and the remaining 15 per cent included acinic cell carcinomas, undifferentiated carcinomas, and papillary adenocarcinomas. There is a 50 per cent likelihood of a lump in the palate in the glandular zone being a cancer. The simplistic diagnosis of a cyst should be considered only after other lesions have been excluded. The recommended form of treatment is surgical excision. Fenestration of the palate is a potential consequence of excision in 30 to 60 per cent of patients, depending upon whether or not the lesion is benign or malignant. The results of surgical treatment are good, but fenestration shoud be accepted as a necessary result of adequate treatment. Surgical treatment of cylindromas continues to be most difficult and demands the utmost of that intangible essence known as surgical judgment. Radiotherapy has been considered as a palliative agent for the treatment of inoperable and recurring cancers, especially cylindromas.


Clinical course of cystosarcoma phyllodes related to histologic appearance.

A series of 17 females having cystosarcoma phyllodes underwent surgical treatment. The subsequent clinical course was observed periodically. Twelve patients had originally histologically benign tumors. Six patients had a recurrence, and of these, malignant characteristics developed in four instances. Five patients had originally histologically malignant tumors. Recurrences were encountered in four. Five patients died from a generalized spread of the tumor. Three of these patients had originally histologically benign tumors, in which malignant features of the recurrent tumor subsequently developed. The surgical treatment ranged from local excision to simple mastectomy with additional roentgenotherapy in three. Neither procedure prevented a recurrence from the original tumor, whether benign or malignant. Poor correlation exists between biologic behavior and histologic appearance of cystosarcoma phyllodes, independent of the surgical procedure. Wide local excision in histologically benign and mastectomy in histologically malignant tumors are advised, but no final decision could be reached. It is proposed to restrict ther term cystosarcoma phyllodes to histologically malignant tumors and use the term fibroadenoma phyllodes in instances of histologically benign tumors.


Effects of dibutyryl cyclic AMP on sinus rate and atrial contractile force in the isolated atrium of the dog.

In blood-perfused isolated canine atrium preparations, single injections of dibutyryl cyclic AMP at a dose range of 1 to 30 mg were made into the cannulated sinus node artery. At doses above 3 mg, dibutyryl cyclic AMP induced a biphasic chronotropic and inotropic response; there was a slight and rapid negative chronotropic and inotropic response, followed by a long-lasting positive chronotropic and inotropic response. The induction of the positive effects was slow, and it took 4 to 8 min to reach a maximum response. These effects continued 20 to 60 min. The ratios of the positive inotropic effect/positive chronotropic effect were very similar to those induced by norepinephrine. In this study, it was confirmed that a large dose of dibutyryl cyclic AMP induced a positive chronotropic and inotropic effect on the canine atrium, and the possibility was discussed that cyclic AMP may be involved in the effects of beta-adrenergic amines on the pacemaker and contractile process of the heart.


The sarcoma arising in giloblastoma: clinico-pathological report of two cases.

Two cases of primary brain tumor composed of an admixture of sarcomatous and gliomatous tissues are reported together with their clinical courses and neurosurgical examination. There is no difference in clinical course nor in the result of examination between the usual glioblastoma multiforme and these admixture tumor. The histological examination at the surgical biopsy and autopsy suggested that enoplastic change of the sarcomatous element occurs markedly on hyperplastic blood vessels in the glioblastoma multiforme. The neoplastic change observed on hyperplastic blood vessels is not affected by radiation therapy.


Responses to exogenous dibutyryl adenosine 3',5'-monophosphate of cardiac output and blood flow in the renal, superior mesenteric and carotid arteries in anesthetized dogs.

Changes in the cardiac output and blood flow in the renal, superior mesenteric and carotid arteries in anesthetized dogs were observed, using the non-cannulating electromagnetic flow meter. Dibutyryl cyclic-AMP, 5 mg/kg body weight, was given intravenously and the following results were obtained: 1) Dibutyryl cyclic-AMP increased the stroke volume and the cardiac output, and slightly increased the heart rate. These effects appeared 3 to 5 min after administration of dibutyryl cyclic-AMP. 2) The mean systemic blood pressure as well as the central venous pressure fell slightly. 3) The renal and the superior mesenteric artery blood flow increased markedly, but the carotid artery blood flow did not change. 4) Distribution of the cardiac output to the renal and superior mesenteric arteries did not change but distribution to the carotid artery decreased. 5) Total peripheral resistance, renal artery vascular resistance and superior mesenteric artery vascular resistance decreased, and carotid artery vascular resistance decreased slightly. 6) The cardiac output and blood flow were enhanced by aminophylline (3 mg/kg), and were not blocked by propranolol (0.3 mg/kg).


Effects of intratracheal instillation of dactinomycin on pulmonary edema and phosphatase activity of the lung lavage fluid in rats.

Intratracheal (i.t.) administration of protein synthesis inhibitors produced pulmonary edema. Of those inhibitors studied, dactinomycin (act. D) was the most potent. Severity of lung damage due to act. D was dose- and almost age-related. Maximal intensity of pulmonary edema was reached on the 3rd day following administration and remained constant for 14 days. Histopathological studies revealed confluent edema of the entire lung. Pretreatment with act D induced tolerance to an LD100 edematogenic dose of thiourea. The effects of i.t. instillation of act. D appear to be localized in the pulmonary tissue. Lung lavage fluid collected from drug-treated rats had higher acid and alkaline phosphatase activities, higher protein content and more leukocyte infiltration than that of control.


Metabolism  of 4-14 C-progesterone in the adult female chimpanzee.

The metabolism of 4- 14-C-progesterone was studied in two adult female chimpanzees (Pan troglodytes). After intravenous injection of 4- 14-C-progesterone, urine was collected for 5 days. The urinary conjugates were hydrolyzed with Glusulase and the extracts purified by chromatography on silica gel, paper and alumina and then crystallized to constant specific activity with or without carrier steroid. The major metabolite in both experiments was pregnanediol which accounted for 39.8% and 49.5% of the recovered dose respectively. Pregnanolone (0.6% and 2.1%) and pregnanetriol (0.1% and 1.5%) were also isolated in smaller quantities. These data suggest that the pattern of progesterone metabolism in the chimpanzee is similar to that of man in that pregnanediol is the major metabolite whereas androsterone is not.


Specific progesterone receptors in human breast cancer.

We have identified a specific progesterone receptor in 11 of 33 human breast cancer cytosols. Since progesterone itself binds to glucocorticoid receptor, to corticosteroid binding globulin (CBG), and to nonspecific components as well as to its own receptor, we have used a synthetic progestin, R5020 (17,21-dimethyl-19-nor-4,9-pregnadiene-3,20-dione), whose binding specificity is restricted to progesterone receptor. Bound R5020 sediments at 8 S in sucrose gradients; binding is competed by excess unlabeled R5020 or progesterone. The receptor is distinct from glucocorticoid receptor and CBG as determined by competition studies using dexamethasone and hydrocortisone. The dissociation constant for R5020 obtained by Scatchard analysis of dextran-coated charcoal assays is approximately 2 times 10- minus 9 M.


Androgen binding to cytosol prepared from epididymides of sexually mature castrated rabbits: evidence for a cytoplasmic receptor.

The presence of androgen-binding activity in cytosol prepared from the major anatomical segments (caput, corpus, and cauda) of the epididymis of castrated sexually mature rabbits has been demonstrated. A portion of this binding activity is likely to be the epididymal androgen receptor. When epididymal cytosol from adult castrated rabbits is analyzed on low-ionic strength (0.01 MKCl) sucrose gradients, two peaks of macromolecular binding could be detected, one congruent to 4.6S and one congruent to 8S. On gradients containing 1.0 M KCl, only one sedimenting form congruent to 4.6S could be demonstrated, suggesting that the 8S component is composed of aggregates. If cytosol was preincubated with labeled androgen, followed by an incubation with unlabeled androgen, and subsequently analyzed for binding on low-ionic strength gradients, only the congruent to 8S peak could be detected, indicating that most of the binding in the congruent to 4.6S region was rapidly dissociable. This suggests that binding in this region was to moieties other than receptor. Since androgen binding proteins (ABP) of testicular origin would have been cleared from the epididymis at the timepoints that we concentrated on for most of these studies, the 4.6S binding probably represents the association of androgen with plasma testosterone binding globulin (TeBG). The binding of androgen to the receptor can be inhibited by cyproterone, while this antiandrogen does not inhibit binding to either ABP or TeBG at the concentration used.


Androgen receptor in human skin fibroblasts. Characterization of a specific 17beta-hydroxy-5alpha-androstan-3-one-protein complex in cell sonicates and nuclei.

Cultured human skin fibroblasts were shown to contain an androgen binding activity (receptor) which was heat-labile and destroyed by trypsin. Specific binding was seen after incubations of these cells with 1,2-3-H-testosterone, 1,2-3-H17beta-hydroxy-5alpha-androstan-3-one (dihydrotestosterone, DHT) and 1,2-3-H-5alpha-androstane-3alpha, 17beta-diol. This receptor had a high affinity (Kd=0,2-1.6 nM) and a high degree of specificity for DHT. It was measured as a 3-H-DHT-protein complex by gel filtration chromatography using a method which distinguishes specific from nonspecific binding. Receptor activity was distributed about equally between nuclear and extranuclear components at all times studied and was present in both compartments when cell incubations were carried out at 4 degrees and 37 degrees. Saturation analysis indicated that there were 1250-18,600 binding sites per whole cell. By sucrose gradient centrifugation the receptor had a sedimentation coefficient (S20,w) of about 4. Cells grown for 8 days without serum in the medium maintained the same levels of 3-H-DHT binding. Within 15 hours puromycin (20 mug/ml) in serum-free medium caused a 40-60 percent decrease in binding for the same cell lines. Although the highest levels of 3-H-DHT binding were observed in fibroblasts from newborn foreskin, appreciable cytosol and nuclear binding were seen in cells from forearm, neck and abdominal skin. Receptor activity was stable during prolonged culture. Fibroblasts from several skin sites from patients with the androgen insensitivity syndrome (testicular feminization) had no detectable specific DHT binding. In this study it was demonstrated that skin fibroblasts can rapidly convert testosterone to its active form, DHT, bind DHT to a specific receptor protein and transport this complex to their nuclei. Therefore this may prove to be a convenient system for studying androgen action in vitro.


Accessory phallic urethra in female pseudohermaphrodite.

The eighth case of a female pseudohermaphrodite with hypertrophy of the clitoris, a perineal and complete accessory phallic urethra, but without adrenal hyperplasia, is reported. The etiology of this syndrome and the rationale of its treatment are discussed and the literature reviewed.


An experimental inactivated vaccine against bluetongue.

Bluetongue virus grown in BHK cells was shown to be inactivated in concentrations of betapropiolactone (BPL)  higher than 0-15 per cent. When the virus, inactivated with 0-2 per cent BPL and prepared as a double emulsion vaccine, was injected into Cypriot sheep, no untoward reactions were observed and neutralising antibodies developed. The antibody titre reached a high level and persisted for at least a year. After re-vaccination, a secondary response was observed. A bivalent vaccine elicited a response to both virus types incorporated. The possibilities of using a polyvalent inactivated vaccine are discussed.


[Cytogenetic mechanisms in the pathogenesis of malignant neoplasms].

Malignant tumors and pre-cancer processes are characterized by a high proliferative activity of cells, high chromosome variability and extremely high genome variability which is 10 times, as high as that in non-tumor actively proliferating tissues. A clear positive correlation is found between the cell division rate and the level of chromosome variability.


"Dog pox": some clinical aspects of an eruptive condition of certain mucous surfaces in dogs.

An eruptive condition seen on certain mucous membranes of dogs occurs frequently but often asymptomatically.  Lesions have been found to be present in over 60 percent of male dogs.  The infection is probably due to a canine herpes virus and has been termed conveniently but inaccurately "dog pox".  In many cases lesions are present without observable clinical signs but severe irritation, especially related to lesions in the lower bowel, can give rise to marked symptoms which may be confused with abnormality of the para-anal sacs, or to chronic diarrhoea of a type suggestive of colitis.  The incidence, sites of infection and possible syndromes which may be ascribed to the condition are briefly discussed on a clinical basis only.


[Plaque characteristics of herpes mamillitis virus (HMV)].

The virus of the bovine herpes mammillitis (VHM) produces a homogeneous plaque population. The plaques are either round or have the form of a slightly irregular circle, cized 2--3 mm in diameter, with a semitranslucent center. The microscopic examination of paticular plaques has revealed a necrotic center composed of destroyed cells, and in cases where the gross examination gives a clearly defined ontour there is a band of both normal and destroyed cells. No morphologic differences have been established in plaque formation so far as local strains KOS/H-ovo and DZS and the reference strain TVA are concerned. It has been found that the number of the plaques formed is directly proportional to the concentration of the inoculated virus. The preciseness and specificity of the plaque reduction test has been demonstrated as well. The comparative testing of hydrolysed starch, agarose, agar, and carboxylmethylcellulose for the plaque study of VHM has shown that best results are obtained with the use of starch and agarose, no morphologic variation being established among plaques overlaid with the four tested substances.


[Electron microscopic study of Mikulicz's cells].

The results are presented of the electron-microscopic examination of the main element of scleroma granulomatous tissue - the Mikulicz cells. New data on the shape and structure of their nuclei as well as on the presence of electron-dense bodies in their cytoplasma are given.


[The effect of microbial contemination of the air on metabolic indices in broiler chickens].

Trials were carried out with two groups of birds, White Plymouth times Cornish crosses. A third, control group of birds was also involved in the experiments, reared by the same technology. The only difference consisted in that the air in the premise of the test birds had a lower microbial content which, during the experiments was 2 to 5 times lower than that in the air of the controls. It was found that under the effect of the higher microbial content the blood serum protein drops like the protein content of the liver and heart musculature, the content of free amino acids rises, the amount of triptophane lowers, and  the activity of GOT and GPT is enhanced. The birds of the control group show higher enzyme activity so far as the glucose-6-phosphatase of the liver is concerned, lower glycogen content of the liver, and lower blood concentration of glucose. By the end of the trials the broilers of the control group were 50 gr less heavy than the lest birds.


[Concurrent vulvovaginitis, respiratory syndrome and keratoconjunctivitis in calves].

Studied was a disease course in calves caused by the IBR/IPV virus with symptoms characteristic of vulvovaginitis, respiratory distrubances and keratoconjunctivitis. Concurrent infections of such type were observed primarily in several-month-old calves (aged from 6--7 weeks to 6 months). In older calves (aged 1--2 years) vulvovaginitis predominated, it being more rarely associated with changes typical of pneumonia, and, as an exception accompanied by keratoconjunctivitis. Dairy cows exhibited predominatly the chronic form of vulvovaginitis. Isolated were 16 strains of the IBR/IPV virus showing properties analogous of those of the standard Oxford strain. Demonstrated were specific IBR/IPV antibodies in calves, cows and bulls (from 24.5 up to 50 per cent). The problem is discussed of the correlation between sterility in cows showing the presence of the IBR/IPV virus and the symptoms of vulvovaginits as well as the presence of hypospermia and balanoposthitis in bulls harbouring the virus infection.


[Histochemical and histoenzymatic study of experimental myocardial infarction in the rat by temporary and permanent ligation of the left coronary artery (author's transl)].

The evolution of experimental myocardial infarction in the Rat with or without revascularization has been studied histochemically and histoenzymatically in 56 animals sacrified after 1, 6, 12, 24, 48 hrs and 7 days. Following permanent ischemis (14 animals), there appeared an extended transversal infarction marked by the complete disappearance of all phosphorylase activity (P-ase) after the first hour. During the first 6 hrs, changes appeared in succinodeshydrogenase (SHD) and cytochrome oxydase (Cyt-Ox). Glucose-6-phosphodeshydrogenase (G6PDH) presisted until lysis of the necrotic focus. It was possible to define a perinecrotic marginal area in which Pase activity is absent and SDH is granular "G3 in nature, characterized by continuous remodeling in the first 48 hrs. Following temporary ischemia (42 animals) the evolution was marked by rapid tissue reactions and early regression of the marginal zones. After 48 hrs and 7 days of survival, the planimetric evaluation of the infarcted area shows a definite reduction in the size of the infarctus in 50% of cases following removal of the ligature after 6 hrs, and in 66% of cases following removal of the ligature after 1 hr. It would appear probable that the revitalization of certain myocardial areas may extend from the marginal zones as is suggested by the reappearance in these zones several hrs after revascularization of P-ase and SDH activity. On the other hand, it is also true that the early restoration of blood flow does not always prevent the occurrence of an extended infarction. Certain recent observations have shown microcirculatory changes which are secondary to anoxia and should be studied further.


The ultrastructure of focal islet cell adenomatosis in the newborn with hypoglycemia and hyperinsulinism.

In a newborn severe persistent hypoglycemia due to an insulin-producing tumorous proliferation of pancreatic islet cells (insulinoma) was observed. The insulinoma showed the histologic pattern of focal adenomatosis of islet cells. According to the present literature the focal proliferation of islet cell complexes seems to be a frequent and particular feature of insulinomas in the newborn. Differential islet cell staining identified 80%-90% of the proliferated islet cells as B cells. 10%-20% of the cells were found to be A or D cells. Ultrastructurally the majority of the proliferated islet cells were well differentiated B cells. The remaining cells represented either A or D cells or a fourth islet cell type with small spheric granules. Electronmicrscopic evidence of transitions between differentiated islet cells, particularly B cells, and the fourth islet cell type suggests that the fourth islet cell type might represent a precursor cell within the APUD-cell system.


[Determination of individual sensitivity of gastric cancer to 5-fluorouracil by tumor tissue respiratory and glycolytic indices].

Eighty one patients were studied for individual susceptibility of gastric tumors to 5-FU as evidenced by endogenic respiration and glycolysis of fresh tumor sections, the main criterion being the suppressed respiration, the accessory one  inhibition of glycolysis. Three groups of tumors were differentiated: susceptible (25.9%), slightly susceptible (28.4%) and unsusceptible (45.7%) to 5-fluoruracil. Mostly adenocarcinomas were found in a group of susceptible tumors, scirrhous and colloid cancers -- in an unsusceptible group. A correlation outlined between the results of determining susceptibility in vitro and clinical data enabled to recommend the method of determining susceptibility to 5-FU for clinical application.


[The clinical picture and diagnosis of mechanical jaundice during tumor growth].

Studies on the clinical course of the lesion in 525 patients with malignant tumors of the bilio-pancreato-duodenal system organs have evidenced that jaundice is not an early sign of the affection. In developed jaundice it is rational to differentiate two main diagnostic stages: presurgical which main task is differentiation between mechanic jaundice and hepatocellular jaundice, the greatest importance being attached to clinical investigation of serum enzymes and clinical data. The second main diagnostic stage is an operation during which all feasible methods of ultimate diagnosis should be employed, including direct cholangiography and urgent microscopy of a tumor.


[The effectiveness of treatment for breast cancer patients].

The study of late results of treatment in 1166 patients with breast cancer permitted to find the percentage of a 5-year survival to be equal to 55.58, 37.93% of patients survived for 10 years, 28.25% -- for 15 years. In the first stage  a 5-year survival was 87.20%, while in the second and third -- 57.7% and 31.56% of patients survived for 5 years, accordingly. The modern methods of radiotherapy (radiosurgical method, interstitial gamma therapy with Au-198 granules, distance grammatherapy on the machine LUCH) enhanced the efficacy of therapeutic measures in patients with widespread breast cancer. A complex diagnosis -- cytology, non-contrast mammography and mathematical analysis by means of a computer contributes to recognition of early forms of breast cancer.


[Surgical treatment of breast cancer patients (according to material from the Rovno, Novgorod, Zvenigorod, Murmansk and Volynsk oncologic dispensaries].

A retrospective analysis of the survival in 230 patients with breast cancer, stage T1N0M0, subjected to mastectomy is presented. The results of treatment are compared with the survival of 62 patients, in whom economic resections were performed (segmental resection, routine mastectomy). In both groups of patients late results of treatment in stage T1N0M0 practically coincided. It was found that about 20% of nonpulpable axillary lymph nodes proved to be involved in metastases. Due to this, it is recommended to perform economic operations simultaneously with surgical dissection of axillary lymph nodes en bloc with a tumor.


Persistent or slow viral infections and related diseases.

The discovery of persistent transmissible agents by veterinarians has led to striking advances in the infectious cause of neuropathies of human beings. There is evidence for persisting infection in congenital rubella and the herpes group of viruses including cytomegalovirus infections. Hepatitis types A and B are candidates for inclusion in the category of persisting viral infections. The rubeola or measles virus is established as a persistent virus which causes elevated antibodies in the serum and cerebrospinal fluid of many patients with severe demyelinating disease such as subacute sclerosing panencephalitis and multiple sclerosis. Elevated antibodies against vaccinia virus have been found in the cerebrospinal fluid of some patients with multiple sclerosis and neuromyelitis optica, a rare form of multiple sclerosis.


[Sudden deafness and cytomegaly virus infection (author's transl)].

A 49-year old patient complained of sudden deafness in his right ear. With the exception of a rising titre level of the complement-fixing antibodies there were no positive findings for the cytomegaly virus. However, the eight-fold rise in the titre level after a latent period made it appear likely that the sudden deafness was caused by a cytomegaly virus infection. This assumption was also supported by the lack of vestibular symptoms, which, according to literature, is quite frequent in virus diseases of the inner ear. The usual therapy led to an improvement in the patient's hearing, which was also accompanied by a drop in the titre level of the antibodies. No prognosis can be made, since similar observations have not yet been recorded.


Cerebrospinal fluid concentration and urinary excretion of cyclic adenosine-3', 5'-monophosphate in various diseases of children.

Cerebrospinal fluid (CSF) concentration and urinary excretion of cyclic adenosine-3',5'-monophosphate(cAMP) were measured in children aged from 3 days to 15 years by the protein-binding method of Gilman (1970). The mean CSF cAMP concentration (22.4 plus or minus 0.6 (S.E.) nmol/l) of 24 "healthy" children tended to be lower (P less then 0.2) than that of adult patients who revealed no pathological findings on clinical examination. No difference in the results was foung between the sexes. High cAMP concentrations were found in CSF of children suffering from cerebellar glioma, hypothalamic precocious puberty, bacterial meningitis, or Cushing's disease. The urinary excretion of cAMP varied from 0.2 to 5.3 in "healthy" and from 1.3 to 7.6 mumol/24 hrs in diseased children. Two children with pheochromocytoma showed a striking decrease in the rate of urinary excretion of the nucleotide after surgical treatment.


Pathogenic mechanisms not operating in Eimeria necatrix infections.

Having investigated certain aspects of Eimeria necatrix coccidiosis in chickens, workers of the Hannover Veterinary School postulated "that death following a single inoculation of a large number of oocysts is due to an alarm reaction and not a specific pathogenic action of the parasites". Because this hypothesis is somewhat revolutionary in its consept, several pieces of evidence on which it is based and several logical deductions which can be made from it have been examined. It has been confirmed that injection of chichens with cysteamine or 5-hydroxytryptamine 30 min before inoculation of the birds with a lethal dose of E. necatrix oocysts reduces subsequent mortality; the reason for this, however, appears not to be the neutralisation of the proposed shock reaction, but rather an inhibition of the excystation process, brought about indirectly through the host. Inoculation of chickens with a non-lethal dose of E. necatrix oocysts 30 min before inoculation with a lethal dose of oocysts was followed by increased mortality rather than the decreased mortality which the hypothesis would predict. Treatment of chickens with sulphadimidine starting 48 h after inoculation resulted in survival of the birds rather than death which would ensue if in fact mortality was due to a shock reaction irreversibly initiated at the time of inoculation. A direct effect of sulphadimidine on the parasite has been shown both in vivo and in vitro.


Alterations in cyclic necleotides in dogs after triiodothyronine.

The effects of triiodothyronine (T3) on plasma and tissue levels (liver, adipose tissue, muscle) of adenosine 3',5'-monophosphate (cyclic GMP) were determined in Mongrel dogs. Plasma cyclic AMP increased to a mean plateau value 165% greater than control values in response to a single intravenous injection of T3 (100-200 mug/kg body weight). This treatment resulted in no increase in plasma cyclic GMP. In liver, cyclic AMP concentration decreased 54%, while cyclic GMP increased 137%. Adipose tissue cyclic AMP levels decreased in control animals during the experimental procedure. On the other hand, animals given T3 had stable or (in one case) increasing adipose tissue cyclic AMP levels. Hence, T3, actually maintained higher levels than that expected, in comparison to the control. Cyclic GMP levels in adipose tissue were not affected by T3. Cyclic AMP and cyclic GMP were unchanged in muscle. In all cases, a time lag occurred (30-45 min) between administration of T3 and subsequent alterations in cyclic nucleotide levels. It was concluded that T3 is capable of altering concentrations of cyclic AMP and cyclic GMP in vivo and that cyclic AMP and cyclic GMP patterns of response are completely different. In liver, a converse relation of the two nucleotides is present. These findings are compatible with the hypothesis that some of T3's action may be explained by its effects upon either cyclic AMP or cyclic GMP.


Glucose inhibiton of the permeation of chloromercuribenzene-p-sulphonic acid in the plasma membrane of pancreatic beta-cells.

The transport of chloromercuribenzene-p-sulphonic acid (CMBS) was studied in microdissected pancreatic islets of ob/ob mice incubated in Krebs-Ringer bicarbonate medium. Glucose inhibited the islet uptake of CMBS. The initial binding of CMBS to superficial thiol groups was inhibited to a lesser extent than was its further permeation into the interior of the plasma membrane. The non-metabolized glucose analogue 3-0-methyl glucose had no effect on the permeation of CMBS. Whereas mannoheptulose, epinephrine and diazoxide lacked any effect in themselves, they tended to counteract the glucose inhibition of CMBS permeation. The ability of glucose to inhibit CMBS permeation is related to its insulin-releasing action rather than to its membrane transport or further metabolism.


Age dependence of plasma oestrogen response to HCG and ACTH in men.

Twelve men, 19-40 years old, and 12 men, 60-86 years old, were given ACTH and in a second test series, HCG. Oestrone, oestradiol-17beta and cortisol were measured in the plasma before and after stimulation. Under basal conditions oestrone and oestradiol as well as the percentage binding of testosterone increased significantly with age, whereas cortisol remained unchanged. The administration of HCG resulted in higher levels of both  plasma oestrone and oestradiol in older subjects than in younger ones, although the percentage increase was smaller. ACTH stimulation leads to higher levels of oestrone in the older group, whereas oestradiol remained unchanged. Cortisol rose 3.5 fold in both groups. Regarding absolute concentrations of oestrone and oestradiol the ability of the adrenals and of the testes to respond to exogenous gonadotrophin and adrenocorticotrophin appeared to be preserved despite ageing, though the relative responses of oestrone and oestradiol after HCG and of oestrone after ACTH were decreased in old as compared to young men.


Effect of diethylnitrosamine on the livers of rats after high oral doses administered at intervals varying between three and twenty-four days.

When 200 mg of diethylnitrosamine per kg of body weight was administered with a stomach tube once a week to female Wistar W.64 rats, all of them died within three weeks from severe hepatocellular injuries and haemorrhages of the liver, lungs and small intestine. Weekly administrations of 100 mg per kg of body weight consistently caused the rats to die after seven to fifteen weeks from severe hepatic cirrhoses characterized by large tubercles. Weekly doses of 50 mg per kg of body weight caused much less severe hepatic cirrhosis. Under these conditions death occurred after 17 to 23 weeks, all the rats having contracted multiple hepatocellular carcinomas and in most cases pronounced hepatic cirrhosis. When this amount of diethylnitrosamine was administered for twelve weeks only the hepatic cirrhosis was less apparent, but a substantial influence on the tumor development was not observed. After a reduction of the dose to 25 mg per kg of body weight once a week all the rats in the relevant group died after 26 to 35 weeks, likewise from multiple hepatocellular carcinomas. Hepatic cirrhosis were then relatively inconspicuous. Similar results were obtained when, not the dose, but the interval between doses (100 mg/kg each) was varied. This clearly differential effect of diethylnitrosamine in the rat thus makes it a simple matter to induce hepatic cirrhosis or cancers that are suitable for a variety of tests and also, for example, as models for therapy trials.


Studies of the plasma kinin-forming system in cirrhosis of the liver.

Kininogen level, that of active kinins and kininase activity in the plasma of patients suffering from cirrhosis of the liver and of healthy people were studied.  The kininogen content was determined by different available methods i.e. the trypsin and acetone techniques and by means of the plasma and glandular kallikrein preparation.  An increase in kininase activity and a lowered kininogen level as determined by all the methods were found in the sick persons.  The maximal decrease in the kininogen level determined by means of the plasma kallikreins is substantiated in connection with the increased fibrinolytic activity of the plasma of the patients with hepatic cirrhosis.


Virus antibody levels in serum specimens from patients with optic neuritis and from matched controls;.

Virus antibody levels in serum specimens taken in acute and convalescent phases from 77 patients with optic neuritits were tested by measles hamagglutination inhibition (HI), measles hemolysis inhibition (HLI), rubella HI, parainfluenza-1 HI, Epstein-Barr immunofluorescence (IF), and against 11 other viruses and mycoplasma pneumoniae with the complement fixation (CF) technique. The virus antibody levels were indicated to be usually very stable, and a fourfold change in virus antibody levels was demonstrated in only eight patients. The virus antibody levels were compared with specimens from two carefully selected control groups. The first control group consisted of 71 healthy persons matched in age, sex and place of residence with the patients with optic neuritis. The other control group consisted of 58 patients with various neurological diseases other than multiple sclerosis (MS) or infectious diseases of the central nervous system. The patients with optic neuritis had significantly higher measles antibody titres than the two control groups in both measles HI and measles HLI tests. Also in 33 patients with optic neuritis of unknown cause, the measles antibody levels were higher than in the control groups. On the other hand, various other antibody tests showed no statistically significant differences between patients with optic neuritis and the control group.


Virus antibody levels in the cerebrospinal fluid from patients with optic neuritis.

Virus antibody levels were studied in the cerebrospinal fluid (CSF) of 58 patients with optic neuritis and 58 control patients with no indication of multiple sclerosis (MS) or infectious disorders of the central nervous system (CNS). The specimens were tested against three different structural components of measles virus with measles hemagglutination inhibition (HI), measles hemolysis inhibition (HLI) and gel precipitation (GP) tests. Measles antibodies occurred in 62 per cent of CSF specimens from patients with optic neuritis, and 21 per cent of the controls. In the specimens from patients with optic neuritis, the positive rate figures were: for rubella HI test 35, parainfluenza-1 HI 16,  and Epstein-Barr virus immunofuorescence (IF) 53 per cent. The frequencies in the control group were 10, 10 and 26 per cent, respectively. Serum/CSF antibody ratios below 80 occurred in measles tests in 45 per cent of patients with optic neuritis and 16 per cent of the control group. Some patients with optic neuritis (but none from the control group) had a reduced serum/CSF antibody ratio in more than one measles antibody test, The patients with optic neuritis had a higher frequency of low serum/CSF albumin ratios indicating blood brain barrier damage, There were, however, several patients with a normal serum/CSF albumin ratio but low serum/CSF immunoglobulin G and measles antibody ratios. This supports the hypothesis that local production of measles antibodies takes place in CNS in some patients with optic neuritis as well as in MS patients. The CSF specimens were further tested against 12 other viruses and mycoplasma pneumoniae complement fixation, but there were no positive specimens. New CSF specimens were taken from five patients during optic neuritis, and from seven patients later on during the follow-up because of the appearance of new neurological symptoms. There were no changes in virus antibody levels, except for two patients with an increase of measles virus antibody titres.


Congenital generalized lipodystrophy.

Six patients with congenital generalized lipodystrophy are described. They had generalized paucity of fat tissue, acanthosis nigricans, prominent superficial veins and muscle hypertrophy. They were mentally retarded. Three had corneal opacities. They had normal external genitalia and none was tall for age. Their bone age was advanced and some had minor skeletal anomalies and nephromegaly. The muscle histology on light microscopy was normal. The majority had elevated serum aldolase and to a lesser degree serum lactic dehydrogenase and creatinine phosphokinase. Four of five examined had a myopathic electromyogram. They had normal or deranged liver function tests. The fatty liver infiltration in one seems to be progressive. Four had a normal and two an abnormal metyrapone test. They had an age-dependent abnormality of growth hormone, insulin and carbohydrate homeostasis.


Preparation of type specific herpes simplex antisera by an immunosorbent method;.

Herpes simplex virus (HSV) type 1 and type 2 specific immune sera have been produced by adsorption with glutaraldehyde-polymerized heterologous virus antigen. The adsorbed sera showed high specificity in complement fixation, immunoelectroosmophoresis and neutralization tests, The precipitin patterns in IEOP indicate the presence of at least five HSV type 1 and three HSV type 2 specific antigens.