Complete necrotization of hepatocellular carcinoma by chemotherapy and subsequent intravascular coagulation: a case report. A 45-year-old man with hepatocellular carcinoma who developed intravascular coagulation following complete tumor regression by chemotherapy is described. After 2 doses of 10 mg of Mitomycin C given into the hepatic artery at the time of selective angiography, and 16 intravenous doses of 5-fluorouracil and Mitomycin C, 2 doses per week, subjective symptoms and hepatomegaly disappeared. Alpha-fetoprotein became negative and a remarkable change in tumor size and vasculature was noted in the arteriogram. Three months after chemotherapy, the patient developed thrombocytopenia, intravascular hemolysis, and acute renal failure. Autopsy disclosed a 8 X 7 X 5 cm solitary, encapsulated hepatocellular carcinoma in the right lobe. The tumor was surrounded by a thick capsule and completely necrotized. Neither intrahepatic invasion nor extrahepatic metastasis was observed. In the kidney, generalized fibrin thrombi were seen in the afferent arterioles of glomeruli as accounted for by intravascular coagulation.


Oat cell carcinoma of the larynx: response to combined modality therapy. A patient with oat cell carcinoma of the larynx with metastasis to cervical lymph nodes is presented. Treatment with radiation and chemotherapy has achieved a sustained remission. Pretherapy staging and combined modality therapy are discussed.


Cyclic nucleotide concentrations in 1,2-dimethylhydrazine and x-ray induced rat small intestinal cancer. The intracellular concentrations of adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5'-cyclic monophosphate (cGMP) were determined in adult male Holtzman rat small intestinal tumors induced by subcutaneous administration of 1,2-dimethylhydrazine (DMH) or X-irradiation of only the hypoxic ileum and jejunum. The levels of cAMP and cGMP in the cancer cells from DMH-treated animals were 50% and 200%, respectively, of the values measured for intestinal tissue. The amounts of cGMP in the lesion of the X-irradiation induced rat small bowel adenocarcinoma determined in the present investigation and of cAMP measured previously were observed to be only 50% of the value for unirradiated intestine. It has thus been shown that in the DMH-induced colon and small intestinal tumors, the X-irradiation induced rat small bowel adenocarcinoma and the human colon adenocarcinoma, the cAMP concentrations are consistently about 50% of the levels measured in comparable normal tissues. These findings of diminished intracellular cAMP concentrations imply a serious cellular defect mechanism occurring in intestinal cancer. However, there appears to be no similar pattern of change for the intracellular concentrations of cGMP, which suggests that different biochemical pathways exist in these malignancies.


Oncogenicity of purine 3-oxide and unsubstituted purine in rats. Injection s.c. of purine 3-oxide into Wistar rats resulted in the appearance of sarcomas and fibromas at the interscapular site of administration, carcinomas in the liver, and a high incidence of s.c. fibromas in the hip at a distance from the site of injection. A small number of liver tumors but not tumors at the injection site appeared in rats to which the parent compound, purine, was administered. Oxidation of purine 3-oxide by xanthine oxidase was found to occur in two steps to yield the potent oncogen 3-hydroxyxanthine. A similar process may occur in vivo since a protein preparation from rat s.c. tissue has similar oxidizing activity.


Effect of structure on tumor specificity of alicyclic alpha-amino acids. The selective affinity of [carboxyl-11C]-1-aminocyclopentanecarboxylic acid (ACPC) for tumor tissue has led us to study the tumor-localizing characteristics of a series of alicyclic alpha-amino acid analogs of ACPC. The tissue distributions of [14C]-1-aminocyclopropanecarboxylic acid, 1-aminocyclobutanecarboxylic acid (ACBC), 1-aminocyclohexanecarboxylic acid, 1-amino-2-methylcyclopentanecarboxylic acid were compared with that of ACPC in Buffalo rats bearing Morris 5123C hepatomas. ACPC and ACBC were found to have significantly higher tumor-to-nontumor concentration ratios than the other four amino acids. ACBC generally had higher tumor-to-nontumor ratios than did ACPC, significantly so for muscle, kidney, and testis and marginally so for blood. These results suggest that [carboxyl-11C]ACBC may be a better agent than [carboxyl-11C]ACPC for tumor imaging by positron tomography.


Growth retardation and prevention of Ehrlich solid tumor by Clostridium perfringens type A spores and culture supernatant. When given by direct s.c. injection into the Ehrlich solid carcinoma 1 week after s.c. tumor transfer, viable crude spores of Clostridium perfringens type A (attenuated mutant strain LNG11 ATCC 29348) inhibited tumor growth and significantly prolonged the life span of male outbred Swiss mice. Under these conditions a concentrated sterile supernatant of a C. perfringens culture proved to be slightly more effective than were viable crude spores. In contrast viable crude spores were ineffective in the treatment of female Swiss mice, but the sterile supernatant retained significant activity. When given at the time and site of s.c. grafting of Ehrlich tumor cells, a concentrated sterile supernatant of a C. perfringens culture prevented tumor growth in 80% of male outbred Swiss mice. Under these conditions viable crude spores prevented tumor growth in 70% of mice and significantly prolonged the life span in the other 30%. When given by i.p. injection and before i.p. grafting of tumor cells, viable crude spores of C. perfringens prevented Ehrlich ascites tumor in 5 of 12 Swiss mice and prolonged life span in the other 7. In contrast concentrated sterile supernatant and viable purified spores were ineffective in the prevention or delay of the growth of Ehrlich ascites tumor. These data indicate that C. perfringens can be a potent antitumor agent without producing the harmful anaerobic infection of solid tumors (clostridial oncolysis.


Mammary tumor virus oncogenesis and tumor immunogenicity in three sublines of the C3H mouse. Mammary tumorigenesis and mammary tumor transplantation immunogenicity have been studied and compared in three sublines of the C3H strain: in standard mammary tumor virus (MTV-S)-infected C3H/He mice; in MTV-S-infected C3H/Ki mice; and in MTV-S-free C3Hf/He mice. The age at the appearance of the first tumor, the growth rate of the tumors in their first transplant generation, and the immunogenicity of each tumor in syngeneic female recipients have been determined for the first tumor to appear in each of 25 breeding females from each of the three sublines. Two statistically significant trends were evident among the tumor characteristics compared in the three sublines: (a) an early appearance of tumors was related to the presence of the MTV-S. The genetically dissimilar sublines, C3H/He and C3H/Ki, both infected with the MTV-S, developed mammary tumors at an average age of about 10 months, 11 months before MTV-S-free C3Hf/He mice; (b) the tumor characteristics of immunogenicity and growth stimulation were related to host genetic factors. The genetically similar sublines, C3H/He and C3Hf/He, developed similar proportions of immunogenic and growth-stimulating mammary tumors; the genetically divergent C3H/Ki subline developed tumors that were not immunogenic and tended to be strongly growth stimulating.


Observations on the question of horizontal transmission of mouse mammary tumor virus. Antigen and tumor incidences in BALB/c and C57BL mice after living as weanlings for 5 weeks in cages with mouse mammary tumor virus-infected females were compared with control BALB/c and C57BL mice living in the same laboratory. All mice were bred continuously, and third-lactation milks were tested for mouse mammary tumor virus antigen by Ouchterlony microimmunodiffusion test. Mammary tumor incidences in the cagemates were not significantly different from those in the controls, although the antigen incidences were significantly greater. However, phosphate-buffered salt solution (0.02 M phosphate, pH 7.4; 0.15 M NaCl; and 0.1% bovine serum albumin) and sham-inoculated mice also had elevated antigen incidences. Repeat tests of milks at the fourth or fifth lactations indicated that more than 50% of those positive at the third became negative at later lactations.


Mutagenesis of Chinese hamster cells in vitro by combination treatments with methyl methanesulfonate and N-acetoxy-2-acetylaminofluorene. Mutational synergism was examined in Chinese hamster V79 cells exposed to methyl methanesulfonate followed by N-acetoxy-2-acetylaminofluorene (AcAAF) at different time intervals. Treatment with 500 micron methyl methanesulfonate resulted in 95% survival of cloning ability and induced approximately 4 azaguanine-resistant mutants/10(5) survivors. Seven micron AcAAF produced 10 times as many mutants, and the survival was 7%. Lethal synergism was observed for methyl methanesulfonate treatments followed by 7 micron AcAAF, and the resulting lethality was unaffected by increasing the time interval between treatments from 1 to 48 hr. However, no significant changes in the mutant frequency from that induced by AcAAF alone were found for treatment intervals of 1 to 63 hr. This result contrasts with the 6-fold enhancement of the AcAAF-induced transformation of Syrian hamster embryo cells exposed to the same combination with a 48-hr interval between treatments, as previously reported (Chem.-Biol. Interactions, 9: 351-364, 1974). The difference in the response of these two cell types demonstrates the difficulties in attempting to extrapolate the known correlation between individual mutagen and carcinogen treatments to combination treatments, with different cell types for the two cellular responses.


Separation and maturation of gonadotrophs from 2A8 clonal cells in vitro. 2A8 clonal cells derived from the epithlium of Rathke's pouch of fetal rats were cultured in growth medium supplemented with fresh rat serum, median-eminence extract and 1-thyroxine. Then, in order to isolate gonadotrophs, cyanogen bromide-activated Sepharose whichwas conjugated with LHRH (LHRH-Sepharose) was added to the culture medium. Fourteen days after incubation of 2A8 cells with LHRH-Sepharose, agranular and granular cells were rapidly bound to LHRH-Sepharose when fresh serum had previously been added in the medium. The cytoplasm of granular cells that were bound to LHRH-Sepharose contained spherical secretory granules (200-250 nm in diameter). These cells were similar in morphology to the FSH and LH gonadotrophs described by Kurosumi (1968). Moreover, many of them appeared as hypertrophied cells that resembled "castration" cells. These results demonstrate that LHRH conjugated to Sepharose can be used to separate gonadotrophs from other 2A8 cells, and it is suggested that the hypertrophy of some cells might be due to persistent stimulation by LHRH which is likely bound to receptors on the cell membrane.


The transfer and stable integration of the HSV thymidine kinase gene into mouse cells. Treatment of mutant mouse cells (Ltk-) deficient in thymidine kinase with Bam I restriction endonuclease-cleaved HSV-1 DNA results in the appearance of numerous surviving colonies which stably express thte tk+ phenotype. Through a series of electrophoretic fractionations in concert with transfection assays, we isolated a 3.4 kb fragment which contains the thymidine kinase gene and which alone is competent in the biochemical transformation of Ltk- cells. In this report, we have examined the distribution of tk sequences in the DNA of several transformed clones following stable gene transfer. A series of complementary experiments involving reassociation kinetics in solution and annealings with tk DNA to restriction-cleaved cellular DNA following electrophoresis and transfer to filters allow us to make the following general conclusions concerning the fate of the tk gene in all clones examined: the tk gene is present in all cells at a frequency of one copy per chromosomal complement; the tk gene is stably integrated in the DNA of all transformants; and integration is not site-specific and occurs at different loci in the DNA of all transformants examined. The existence of a single active tk gene in tk+ transformants now facilitates an analysis of the sequence organization of tk- mutant cells and provides a useful model system for studies on the transfer of cellular genes.


Mutants of SV40 with an altered small t protein are reduced in their ability to transform cells. Mutants of SV40 with deletions of various sizes mapping between 0.54 and 0.59 on the genome grow at a rate equal to or slightly slower than that of wild-type virus, in a range of host cells. Their ability, however, to induce transformation in several mouse, rat and rabbit cell lines is impaired. The extent of transformation observed is dependent upon the assay used to measure it, but in general, the ability of the mutants to transform falls as the size of the deletion increases. In addition, rat embryo fibroblasts transformed by deletion mutants have fewer of the characteristics of a fully transformed phenotype (for example, growth in low serum, increased saturation density, growth in semi-solid medium) than those transformed by wild-type virus. During lytic infection, immunoprecipitable T antigen produced by the deletion mutants is of the same size as that seen during infection with wild-type virus, and is present at a similar level. Mutant virus-coded small t protein, however, is reduced in size compared with that from wild-type virus. For each mutant, the reduction in protein size is dependent upon the amount of DNA deleted, but not on the relative position of the deletion in the genome. These results demonstrate that the DNA sequences mapping between 0.54 and 0.59 on the viral genome code for the small t protein, and that SV40-induced transformation is at least partially dependent upon the expression of this protein.


Complete sequences of the ribosome recognition sites in vesicular stomatitis virus mRNAs: recognition by the 40S and 80S complexes. Nucleotide sequences of the ribosome-protected translation initiation sites from the vesicular stomatitis virus (VSV) M and L protein mRNAs have been determined, completing the sequences of the sites from all the VSV mRNAs. A low level of protection at two internal AUG-containing sites in the N mRNA is also described. Small homologies are evident among some of the sites, but there are no obvious features common to all the sites other than a single AUG codon. In contrast, a large homology between the VSV M mRNA site and the alfalfa mosaic virus coat mRNA site (Koper-Zwarthoff et al., 1977) is noted. This homology suggests the existence of a common ancestral gene for these two apparently unrelated viruses. For each VSV mRNA species, the smallest sites protected in either the 40S or 80S initiation complexes are identical. These sites always contained the initiation codon, but only contained the capped 5' end in those mRNAs having the 5' end near the initiation site. If 40S ribosomes bind to the capped 5' end, either they do not protect it from nuclease digestion or the protection is only transitory in some VSV mRNAs. Consideration of the structures of the ribosome binding sites suggests that the differential effects of hypertonic shock on translation (Nuss and Koch, 1976) may be related to the distance between the 5' end of the mRNA and the initiation codon.


Murine interferon system regulation: isolation and characterization of a mutant 3T6 cell engaged in the semiconstitutive synthesis of interferon. We describe the isolation and characterization of a virus-resistant mutant of murine 3T6 cells. The mutant, designated 3T6-VrB2, displays a high degree of resistance to infection by members of the toga-, rhabdo- and picornavirus classes. The level of this resistance to infection is similar to the parent 3T6 pretreated with approximately 100 lU/ml of interferon. Upon co-cultivation of 3T6-VrB2 cells with interferon-sensitive mouse cells, an antiviral state is induced in the latter cells as measured by a reduction of virus yield following infection. The nature of the induction is defined by a series of experiments using anti-mouse interferon antiserum. In the presence of this antiserum, the ability of the mutant to induce an antiviral state in interferon-sensitive mouse cells upon co-cultivation is eliminated. Additionally, growth of the mutant cells in the presence of this antiserum causes a reversal of the virus-resistant phenotype. Our results indicate that 3T6-VrB2 contains a mutation affecting the regulation of the murine interferon system such that the cell is engaged in the semiconstitutive synthesis of interferon.


Chemotaxis in Dictyostelium discoideum: effect of concanavalin A on chemoattractant mediated cyclic gmp accumulation anlight scattering decrease. In cells of the cellular slime mold Dictyostelium discoideum concanavalin A (Con A), at a concentration of 100 microgram per ml, inhibits folic acid and cyclic AMP induced decrease in light scattering. Con A has no effect on folic acid mediated cyclic GMP accumulation and increases cyclic AMP mediated cyclic GMP accumulation two-fold. At a lower Con A concentration, 10 microgram per ml, changes in light scattering induced by folic acid are normal and cyclic AMP induces a monophasic instead of a biphasic response. The stimulatory effect of Con A on cyclic AMP mediated cyclic GMP accumulation is still observable at 10 microgram Con A per ml. When cells are repeatedly stimulated with cyclic AMP, a decrease in light scattering without being accompanied by changes in cyclic GMP concentration is observed. Based on these results a model for chemotaxis is proposed.


[Effect of synthetic ACTH (beta 1-23 corticotropin) on gestation and parturition in rats]. Daily administration of synthetic ACTH (betaI-23 corticotrophine) from day 15 of pregnancy induces delay or inhibition of parturition in normal or ovariectomized Rats injected or not with estradiol. Parturition takes place at approximatively normal time, when ACTH is stopped and estradiol continued. Radio-immunological assays show an increase of blood progesterone level after ACTH administration in castrated, but not in castrated and adrenalectomized Rats. Mechanisms of ACTH action are discussed.


[Aflatoxin is present in primary liver cancers in residents of Zaïre]. Aflatoxin is localized by fluorescence microscopy in primary liver cancer in man from Zaïre.


[Studies of spin-labeled spectrin].

[Studies of spin-labeled spectrin]. Spectrin isolated from human erythrocytes has been spin-labeled with five maleimide nitroxides. The mobility of the labels is strongly dependent on their size, and on the temperature. A thermal transition of spectrin is shown to occur above 30 degrees C. Calcium and magnesium provoke a strong immobilization of the labels. This effect is yet more pronounced when spectrin is allowed to reassociate with the cytoplasmic surface of the membrane.


[Loss of steroidogenesis in a mouse adrenal cell line after simian adenovirus 7 (SA7) transformation]. After transformation by the simian adenovirus 7 (SA7), the Y-1 Mouse adrenal tumor cells are no longer able to produce steroïds either spontaneously, or after specific stimulation. Nevertheless, these stimulations are able to increase the level of cAMP both in transformed and original cell line. The differentiation is associated with an intracellular modification of steroïdogenesis which seems to be localized in the metabolic chain of steroïd synthesis after the increase of the cAMP level.


[Relations between malaria and Burkitt's lymphoma]. Malaria, especially forest malaria, is the environmental factor which creates endemic conditions for the Esptein-Barr virus related Burkitt's lymphoma. The Plasmodium seems active through a mitogenic factor rather than through the so often invoked immunological depresssion.


[Modulator role of prostaglandins (Pg) in adrenergic neurotransmission in carotid sinus]. In Rabbit carotid sinus, the presence of sympathetic nerve endings capable of releasing noradrenaline has been demonstrated. The release of NA in response to sympathetic nerve stimulation was decreased by PgE2 and a precursor of Pg (arachidonic acid) but was strongly increased by an inhibitor of Pg biosynthesis (indomethacin). The experiments carried out demonstrated that freshly synthesized Pg acts in the same way as exogenous Pg and suggested that Pg could have a regulating effect on adrenergic neurotransmission in carotid sinus. The role of this regulating mechanism in the physiology of carotid sinus has been discussed.


[Inhibitory effects of LHRH on ovarian LH receptors in rats]. The administration of LHRH or of its analog [D-Ala6, des-Gly-NH10(2)] LHRH ethylamide to the female rat on day 10 of gestation resulted in termination of pregnancy and in a marked decrease in ovarian LH receptors and plasma progesterone level. Asingle injection of either compound during diestrus I also led to a decrease in LH receptors. These data demonstrate the high sensitivity of the control of ovarian LH receptors by circulating gonadotrophins and suggest that the contraceptive action of pre- and postcoitally administered LHRH is mediated by negative control of LH receptors.


[Role of alpha and beta adrenoceptors in the regulation of adrenergic neurotransmission in the carotid sinus]. The experiments carried out showed the presence- in sympathetic nerve endings ot the carotid sinus- of alpha and beta adrenoceptors which by means of respective negative and positive feedback processes, modulated NA release induced by a sympathetic nerve stimulation. Similarly, Pgs acted by means of a negative feedback mechanism to regulated adrenergic neuro-transmission in carotid sinus but they could not be considered as the chemical mediators of either alpha or beta adrenoceptors.


[Effect of dibutyryl cyclic AMP on cerebral energy charge and electrocorticographic activity in rats under asphyxic hypoxia]. In Rats anesthetized with pentobarbital and submitted to brief asphyxic episodes, intracerebroventricular administration of dibutyryl cyclic AMP inhibits the decrease of energy charge, delays the disappearance of electrocorticographic activity (E Co G) and shortens the recovery time of E Co G.


[Murine type C endogenous virus expression in murine 129 teratocarcinoma cell line]. Murine type C viral information is detectable in the cellular genome of both differentiated and undifferentiated cell lines derived from 129 Mouse teratocarcinoma. Cytoplasmic RNA expression which is negligible in differentiated cells, is significantly higher in multipotential undifferentiated cells. Furthermore it was observed that in vitro differentiation of multipotent cells leads to a decrease of this expression.


[Inhibitory effects of LHRH on LH receptors in the rat testis]. A single injection of LHRH to the adult male rat, as of its analog [D-Ala6, des-Gly-NH102] LHRH ethylamide, resulted in a marked decrease in LH receptors in the tests. Plasma testosterone level and the weight of the seminal vesicles and prostate were also decreased after treatment. These data demonstrate that LHRH can decrease the sensitivity of LH receptors and testicular function in the rat.


[Absence of lipolytic action of adrenaline on a human subcutaneous adipose tissue. Role of alpha-adrenergic receptors]. Epinephrine cannot stimulate adipocytes' lipolysis of human subcutaneous adipose tissue (lateral part of the thigh) while a clear lipolytic action can be shown on the omental tissue. However, at the same concentrations, isoproterenol (a beta-agonist) exerts a strong adipokinetic effect on adipocytes of the both types of adipose tissue. An alpha-adrenolytic (phentolamine) enhances the lipolytic action of epinephrine on the omental adipose tissue and unmasks a lipolytic action of epinephrine on the subcutaneous. Epinephrine antagonizes the lipolysis induced by theophyline on the subscutaneous adipocytes, this action is increased by propranolol (a beta-adrenergic blocking agent). The unresponsiveness to epinephrine of the subcutaneous adipose tissue studied here could be linked to a strong antilipolytic alpha-adrenergic effect.


[Analytical morphodynamics. Practical applications]. The analytical morphodynamic method, based on the study of the fate of the biogic units of an organ at the level of well or poorly developed lesions, has been applied to the analysis of chronic obliterating arteriopathies of the extremities, of benign tumors of the mammary parenchyma, and of malignant hepatomas. In morbid states as different as a reactive process and a benign or malignant neoplastic process, it allowed to reconstritute the histogenesis of complex tissue changes and led to interesting prospects, in the field of general biology and pathogenesis.


The interaction of human plasma glycosaminoglycans with plasma lipoproteins. II. Hemagglutination studies. Formalinized, tannic acid-treated sheep erythrocytes coated with low density lipoproteins (LSL) or apoprotein B (apo-B) are are agglutinated by anti-apo-B immunserum. Those coated with high density lipoproteins (HDL) or apoprotein A-I(apo-A-I) are agglutinated by anti-apo-A-I immunserum. These coated formocells have been used to study the interactions of lipoproteins and apoproteins with plasma glycosaminoglycans (GAG). The sulfate-rich species of plasma GAG agglutinates cells coated with LDL, HDL, apo-B, and apo-A-I at ionic concentrations above 0.15 M. The less-sulfated species of plasma GAG does not agglutinate the coated cells but inhibits the agglutination caused by the sulfate-rich species. Treatment of the sulfate-rich GAG with papain causes a reduction in molecular weight by one-half and also causes a loss of its agglutinating activity. These results suggest that the sulfate-rich plasma GAG, consisting of two glycan chains linked to a peptide backbone, cause agglutination by binding to two or more formocells. In contrast, the less-sulfated plasma GAG, consisting of single, short glycan chains, are incapable of causing agglutination but may prevent it by covering specific binding sites present on the coated cells.


The estimation of pyrophosphate in urine with uridine-5'-diphosphoglucose pyrophosphorylase. The concentration of pyrophosphate in urine has been determined using the enzyme uridine-5'-diphosphoglucose pyrophosphorylase. The technique is relatively quick and easy to perform. Reproducibility of results was good, and results from recovery experiments were excellent. The mean concentration of pyrophosphate found in early morning urine samples from 14 normal adults was identical with that reported by other workers using an ion-exchange method to measure pyrophosphate. The pyrophosphate concentration was related approximately to the phosphate concentration.


Apolipoprotein A-1 in cholestatic liver diseases. The apolipoprotein A-1 in cholestatic liver diseases was determined by rocket immunoelectrophoresis. A decrease in the serum apo A-1 was generally observed in LP-X positive liver disease, particularly in the patients with extrahepatic biliary obstruction. With reference to LP-X levels and LCAT activity, the results indicated that not only hepatocellular damage but also cholestasis contribute to the decrease of apo A-1 concentration in patients with liver disease.


Pathology of salivary glandular tumours. The incidence and pathological features of the various tumours arising from salivary glandular tissues are described in the light of the World Health Organization classification of these tumours. The modern concepts relating to their histogenesis are outlined and some of the difficulties which may be encountered in their histological diagnosis are also discussed.


Heterogeneity of the clinical syndrome in patients with systemic lupus erythematosus and genetic deficiency of the second complement component. Two patients with systemic lupus erythematosus associated with homozygous deficiency of the second complement component (SLE-C2D) illustrate the different clinical disease patterns found in patients with this illness. Despite the differences in extent and severity of clinical manifestations and serological findings, the renal disease was similar and kidney function was well preserved in both patients. Renal microscopic changes were focal and segmental, deposits of immuno-globulins and complement components were present by immunofluorescent staining, and dense deposits were seen by electron microscopy. Tubulo-reticular inclusion bodies were found in glomerular endothelial cells and lymphocytes of both patients, but not in the lymphocytes of a clinically healthy C2D sibling. The findings in these two patients stress the importance of careful evaluation to determine the presence of systemic disease in patients with SLE-C2D and suggest that an intact classic complement pathway is important in the development of severe lupus, nephritis, but is not needed in the pathogenesis of lupus skin lesions.


Viral infections in pregnancy.

Viral infections in pregnancy. The responsibility of the physician for viral infections in pregnancy has changed dramatically in the last 15 years. Genital herpes infections have become much more frequent, and the recognition of these infections in women near term is particularly important. Cytomegaloviruses have been found to be the most commom cause of congenital infection, but no satisfactory methods are available for prevention or treatment. Varicella is now recognized as a teratogen when infection occurs in the first 4 months of pregnancy. It can also cause severe generalized disease in the newborn when infection takes place in the last few days of gestation. Hepatitis B infections are becoming more frequent in many populations, and this virus can transmit to the child, particularly when maternal infection occurs in the last trimester. Venezuelan equine encephalitis virus has been found to be a teratogen. This must be considered in southern states such as Florida and Texas, as well as in Central and South America. Rubella, fortunately appears to be coming under control with the intensive use of serodiagnosis and vaccines in the United States.


An outbreak of the conjunctival form of infectious bovine rhinotracheitis. An outbreak of severe conjunctivitis involved approximately 30% of a dairy herd of 118 cattle. Infectious Bovine Rhinotracheitis (IBR) virus was identified by cytopathic effects and fluorescent antibody techniques from conjunctival swabs of affected animals. The disease typically presented as a unilateral ocular disease with systemic signs of pyrexia, inappetance, decreased milk production, and some subsequent abortions.


Estimating adrenal cortical function in dogs with ACTH. The peripheral blood response to intramuscular injection of 10 units ACTH in dogs was investigated because no experimental evidence for the standardization of this procedure for clinical use was available. Following the injection of ACTH in sodium chloride solution, neutrophilia, monocytosis, eosinopenia, and lymphopenia occurred. With the exception of eosinopenia, the greatest change in the concentration of each cell type in peripheral blood occurred between 2 and 4 hours post injection. The maximum change in eosinophil numbers occurred between 4 and 6 hours post injection. When all cell types were considered, 4 hours post injection was the most suitable time to measure the cellular response in peripheral blood in dogs which respond to ACTH. The data indicate that change in the ratio of neutrophils to lymphocytes (N/L) prior to and at 2 to 4 hours after ACTH injection in normal dogs was a sensitive index of response and occured sooner than eosinopenia. The extent of change in the N/L ratio was such that accuracy in interpretation could be obtained by counting as few as 40 cells.


Cytologic examination and viral and bacterial culture in herpes simplex, herpes zoster, and varicella. Cytologic examination of epithelial cells from the base of vesicles, virus isolation, and bacterial culture were carried out in thirty-one patients with herpes simplex, in eleven patients with herpes zoster, and in three patients with varicella. Determination of herpes simplex complement fixation reaction was made in the patients with herpes simplex. Cytologic manifestations consistent with herpes were found 65 percent of patients with herpes simplex, while herpesvirus hominis was isolated in 77 percent of these patients. Diagnostic cytologic manifestations were found in 82 percent of the patients with herpes zoster or varicella. Varicella-zoster virus was isolated in 27 percent of these patients. The presence of pathogenic bacteria did not seem to influence the frequency of virus isolation or finding of characteristic cytologic features.


Behaviour of the mitochondrial respiratory chain in vivo. The interrelation of neuronal function and oxidative metabolism of the brain is most incisively studied by optical techniques in vivo. When O2 becomes limited the respiratory chain becomes reduced, extracellular potassium activity is increased, the EEG is depressed and excitability declines, all as expected. Under mildly hyperoxic conditions however, the opposite responses occur, which indicate an absence of a critical tissue Po2 and a continuum of dependence on the O2 concentration, including levels well above the limits for maximal activity of the respiratory chain in isolated mitochondria in vivo. The unexpectedly high steady state of reduction of cytochrome a, a3 provides the basis for a new hypothesis of an extra energy conservation site between cytochrome a3 and O2 and leads to a consideration of a reaction mechanism of O2 with four electrons and four H+ ions occurring at an enzymically active centre of cytochrome a, a3. A special function of the cytochrome a, a3 complex is implied in regulating cellular K+ transport and thus excitability.


Ambulatory detoxification of heroin addicts: a follow-up study. The present study was devised to investigate whether or not 18 psychosocial characteristics of 141 male and 43 female heroin addicts who entered a 21-day ambulatory detoxification program were correlated with (1) length of stay, (2) completing the full course of detoxification, (3) being successfully transferred to aftercare treatment, and (4) being locatable for follow-up one month later. The study also described the status of the patients who were locatable one month after receiving their last dose of methadone in the detoxification program. Only 74 persons were contacted one month after receiving their last dose of methadone, and they were all in treatment. Eighty-three addicts were transferred to other programs before leaving the detoxification program. There were only two significant relationships between the 18 psychosocial characteristics and the four dependent variables. Number of arrests was inversely related to length of stay, and whites were more likely to be transferred to an aftercare program than blacks. The implications of the results for evaluating other ambulatory detoxification programs and for improving retention rates were discussed.


[Comparative therapeutic investigations of pivmecillinam and amoxycillin (author's transl)]. The therapeutic value of pivmecillinam and amoxycillin in urinary tract infections was compared in a double blind study. No therapeutic superiority or inferiority of either substance could be demonstrated. The study shows that a therapeutic comparison of two antibiotics using everyday treatment is possible. Before the introduction of new antibiotics at least equal effectiveness of the substance should be documented with appropriate investigations.


Adrenal contribution to circulating estrogens in woman. To study the contribution of adrenal glands to circulating estrogens in woman, the concentrations of estrone (E1), estradiol-17 beta (E2), and estriol (E3) in adrenal and peripheral venous blood were measured by radioimmunoassay and the grandular secretion of estrogens after ACTH stimulation was investigated by analyzing the adrenal vein levels of these steroids in patients with breast cancer who were undergoing a therapeutic adrenal operation. Furthermore, adrenal secretion rates of estrogens and cortisol were estimated. It was shown that there existed greater concentrations of estrogens in adrenal vein than in peripheral blood; about 3 times higher for E1 (p less than 0.001), and 2 times higher for E2 and E3 (p less than 0.05). Administration of ACTH caused a significant increase of E1 and E2 concentrations in adrenal venous blood to mean 150% of the basal levels that was comparable to the increase of cortisol. Apparent adrenal secretion rates of estrogens estimated under surgical situation were calculated to be 7.7 +/- 1.7 (M +/- SE) microgram/day for E1, 1.9 +/- 0.3 microgram/day for E2, and 0.3 +/- 0.2 microgram/day for E3, while the secretion rate of cortisol was 52.7 +/- 8.2 microgram/min. The present study demonstrates that the direct adrenal secretion of not only E1, but E2 and E3 contributes to the circulating estrogen levels, and it is suggested that the adrenal glands might be responsible for the relatively important source of estrogen production in the aged woman.


Induction of mammary prolactin receptors and lactose synthesis after ovariectomy in the pregnant mouse. The development of prolactin receptors in the mammary gland after ovariectomy was investigated in pregnant KA mice. Mice were ovariectomized on day 13 of pregnancy and used for the determination of the amount of specific binding of 125I-labelled prolactin to the mammary tissue, and the contents of lactose and nucleic acids in the mammary gland 0, 8, 24, and 72 hr after the operation. The specific binding of 125I-labelled prolactin, lactose and RNA contents in the mammary gland remained low until 8 hr, sharply increased 24 hr and decreased 72 hr after ovariectomy. When ovariectomized mice were treated with 0.2 mg progesterone, pregnancy was maintained and an increase (1.5-fold) in the amount of specific binding was observed with an increase of lactose content. Five mg progesterone completely inhibited lactose synthesis. Cortisol administered with progesterone did not show any specific change at the dose used (0.5 to 10 mg). Although the amount of specific binding was also increased after hysterectomy, this increase (2-fold) did not fully cover the increase after ovariectomy (3-fold). These results suggest that the recepter site for prolactin is induced before the initiation of lactose synthesis caused by ovariectomy during pregnancy.


[The ACTH content of plasma in fetal and newborn swine]. ACTH concentration has been estimated radioimmunologically in fetal plasma (100th day of gestation) and in plasma of newborn piglets during the first 24 hours of life and in sows. In comparison to the values of ACTH in sows at the 100th day of gestation during anaesthesia (175 pg/ml) and sows at parturition (235 +/- 77 pg/ml) the concentration in fetal (558 +/- 163 pg/ml) and newborn piglets (448 +/- 158 pg/ml) was much higher. On an average ACTH concentration increased during the first 24 hours of life up to 998 +/- 628 pg/ml. The results are compared to those in other species.


Effect of gonadotropic hormones on the concentration of adenosine 3', 5'-monophosphate in the rat pineal organ. Concentration of adenosine 3',5'-monophosphate (cAMP) was investigated in the male rat pineal organ after a treatment with human chorionic gonadotropin (HCG). HCG significantly decreased cAMP concentration in the pineal organ.


Interchangeable copper and iron proteins in algal photosynthesis. Studies on plastocyanin and cytochrome c-552 in Chlamydomonas. The interrelation of the copper protein plastocyanin, and a soluble c-type cytochrome, c-552, in photosynthetic electron transport has been studied in the genus Chlamydomonas. With C. reinhardtii the plastocyanin: cytochrome c-552 ratio could be changed from 300:1 less than 1:16 simply by omitting copper from the medium, without any other detectable change. Plastocyanin was indetectable in a second species, C. mundana, for which the cytochrome c-552 level was always very high. The properties of Levine's C. reinhardtii mutant lacking plastocyanin, ac-208, were studies and it was found that the photosynthetic capabilities of a suppressed phenotype and suppressed genotype could be explained by reference to the cytochrome c-552 levels. Both proteins were successfully used in reconstitution experiments with chloroplast fragments. Both showed very fast kinetics for reduction by purified Chlamydomonas cytochrome f, but the rate of electron transfer from one to the other was much slower. It is concluded that they constitute an interchangeable pair, and the rationale for this and possible analogies are both discussed.


Adenosine 3':5'-monophosphate in perinatal rat liver. Ontogeny and response to hormones. Developmental changes in the concentration of adenosine 3':5'-monophosphate (cyclic AMP) and the effects of glucagon and epinephrine were studied in the perinatal rat liver. Hepatic cyclic AMP concentration doubled during the last day of gestation. After birth, the cyclic AMP concentration continued to increase and maximal levels were observed on the fifth postnatal day. Surgical delivery of foetuses on days 20, 21 and 22 of gestation resulted in a rapid increase in cyclic AMP concentration. Maximal concentrations were reached within one hour of delivery in the day-21 and day-22 foetuses. However with surgically delivered day-20 foetuses, the cyclic AMP concentration increased for a least two hours. Glucagon and epinephrine increases the hepatic cyclic AMP concentration in rats delivered surgically on days 20, 21 and 22 of gestation and in postnatal rats. Maximal stimulation by epinephrine was observed in 2-day-old rats. Maximal stimulation by glucagon was observed in 10-day-old rats. The results support the hypothesis that cyclic AMP is the intracellular effector for the synthesis of some enzymes in the perinatal rat. The cyclic AMP concentration in the perinatal rat liver in vivo appears to be controlled by changes in the relative concentrations of plasma glucagon and insulin.


Identification of some metabolic products of 5' -deoxy-5' -S-isobutylthioadenosine, an inhibitor of virus-induced cell transformation. 5' -Deoxy-5' -S-isobutylthioadenosine (iBuS)5' Ado has been shown to be rapidly degraded to 5-deoxy-5-S-isobutylthioribose and adenine in procaryotes. In chick embryo fibroblasts there are two metabolic pathways for (iBuS)5' Ado degradation: (a) oxidative deamination into 5' -deoxy-5'-S-isobutylthioinosine (the main product) and (b) hydrolysis into 5-deoxy-5-S-isobutylthioribose plus adenine. The latter reaction is not due to bacterial contamination, since the same results were obtained under sterile conditions and in chick embryo fibroblasts in culture. The inhibition of the virus-induced cell transformation reported by us previously was due to (iBuS)5' Ado rather than to the main metabolic product of this molecule in chick embryo fibroblasts.


An intramolecularly quenched fluorescent tripeptide as a fluorogenic substrate of angiotensin-I-converting enzyme and of bacterial dipeptidyl carboxypeptidase. The N-acyltripeptide 2-aminobenzoylglycyl-p-nitrophenylalanylproline was synthesized and applied as a substrate in the assay of angiotensin-I-converting enzyme from calf lung and human serum, and of the bacterial dipeptidyl carboxypeptidase from Escherichia coli. This compound belongs to a new class of substrates for proteolytic enzymes, having the general structure F--X--Q in which fluorescence of group F  is quenched by intramolecular interaction with the group Q. Enzymatic cleavage of the peptide chain (X stands for one or more amino acid residues) generates the unquenched F-containing derivative and the resulting fluorescence is used for quantitative measurement of the hydrolysis rate. Cleavage of the Gly-Phe(NO2) peptide bond in the weakly fluorescent 2-amino-benzoylglycyl-p-nitrophenylalanylproline molecule results in appearance of the 71 times higher fluorescence (lambdamax = 415 nm) of 2-aminobenzoylglycine. Continuous recording of the rising fluorescence allows convenient, sensitive and specific determination of the enzymatic activity, applicable to crude enzyme preparations and human serum. The activity of the mammalian enzyme, measured by this method, is enhanced by Cl- ions and inhibited by low concentrations of EDTA and [Asn1, Val5]angiotensin II. Kinetic measurements showed Michaelis-Menten behavior, Km = 0.21 +/- 0.1 mM and 0.16 +/- 0.1 mM for the calf lung and the bacterial enzyme respectively.


The regulation of glycogen metabolism. Purification and characterisation of protein phosphatase inhibitor-1 from rabbit skeletal muscle. Inhibitor-1  is a protein which inhibits phosphorylase phosphatase only when it has been phosphorylated by cyclic-AMP-dependent protein kinase [Huang, F. L. and Glinsmann, W. H. (1976) Eur. J. Biochem. 70, 419--426]. Inhibitor-1 was purified by a heat treatment at 90 degrees C, precipitation with ammonium sulphate, chromatography on DEAE-cellulose, gel filtration on Sephadex G-100, and finally rechromatography of the phosphorylated protein on DEAE-cellulose, The protein was purified 4000-fold and 1.5 mg per 1000 g muscle was obtained in seven days corresponding to an overall yield of 15-20%. The purified protein was in a state approaching homogeneity as judged by the criteria of polyacrylamide-gel electrophoresis and ultracentrifugal analysis. The concentration of inhibitor-1 in vivo was calculated to be 1.5 micron, which is at least as high as the concentration of phosphorylase phosphatase. The amino acid composition of inhibitor-1 showed several unusual features. Glutamic acid and proline accounted for nearly one third of the residues, tyrosine, tryptophan and cysteine were absent, and the content of aromatic amino acids was very low. The molecular weight measured by sedimentation equilibrium centrifugation was 19200 and by amino acid analysis was 20800. These values were lower than the mol. wt 26000 determined empirically by gel electrophoresis in the presence of sodium dodecyl sulphate, and much lower than the apparent molecular weight of 60000 estimated by gel filtration on Sephadex G-100. The gel filtration behaviour, stability to heating at 100 degrees C and amino acid composition suggest that inhibitor-1 may possess little ordered structure. The phosphorylated from of inhibitor-1 contained close to one molecule of covalently bound phosphate per mole of protein, which is consistent with the previous finding of a unique decapeptide sequence at the site of phosphorylation, Ile-Arg-Arg-Arg-Arg-Pro-Thr(P)-Pro-Ala-Thr- [Cohen, P., Rylatt, D. B. and Nimmo, G. A. (1977) FEBS Lett. 76, 182-186].the phosphorylated form of inhibitor-1 inhibited phosphorylase phosphatase activity (0.02U) by 50% at a concentration of only 7.0 nM in the standard assay, but the phosphorylated decapeptide was 1000-2000 times less effective as an inhibitor.


The regulation of glycogen metabolism. Phosphorylation of inhibitor-1 from rabbit skeletal muscle, and its interaction with protein phosphatases-III and -II. Inhibitor-1 from rabbit skeletal muscle was phosphorylated by protein kinase dependent on adenosine 3' :5'-monophosphate (cyclic AMP), but not by phosphorylase kinase or by glycogen synthetase kinase-2. Protein phosphatase-III, isolated and stored in the presence of manganese ions to keep it stable, was in a form which catalysed a rapid dephosphorylation and inactivation of inhibitor-1. The kinetic constants for the dephosphorylation of inhibitor-1 [Km = 0.7 micron, V(rel) = 40] were comparable to those for the dephosphorylation of phosphorylase kinase [Km =1.1 micron, V (rel) = 62] and phosphorylase [Km = 5.0 micron, V (rel) = 100]. The dephosphorylation of inhibitor -1 was inhibited by inhibitor-2, indicating that it was catalysed by protein phosphatase-III, and not by another enzyme that might be contaminating the preparation. When protein phosphatase-III was diluted into buffers containing excess EDTA, it lost activity initially, but after 90 min, the activity reached a plateau that remained stable for at least 20h. The initial loss in activity varied with the substrate that was tested; it was 20-30% with phosphorylase a, 50-60% with phosphorylase kinase and greater than or equal to 95% with inhibitor-1. This form of protein phosphatase-III was inhibited by inhibitor-1 in a noncompetitive manner, and the Ki for inhibitor-1 was 1.6 +/- 0.3 nM. The phosphorylase phosphatase, phosphorylase kinase phosphatase and glycogen synthetase phosphatase activities of protein phosphatase-III were inhibited in an identical manner by inhibitor-1. This result emphasizes the potential importance of inhibitor-1 in the regulation of glycogen metabolism, since it can influence the state of phosphorylation of three different enzymes. The formation of the inactive complex between inhibitor-1 and protein phosphatase-III was reversed by incubation with trypsin (which destroyed inhibitor-1, but not protein phosphatase-III) or by dilution of the inactive complex. Kinetic studies, using the form of protein phosphatase-III which dephosphorylated inhibitor-1 very rapidly, demonstrated three unusual features of the system: (a) inhibitor-1 was still as powerful and inhibitor of the dephosphorylation of phosphorylase a and phosphorylase kinase a even under conditions where it was being rapidly dephosphorylated; (b) inhibitor-1 was not an inhibitor of its own dephosphorylation; (c) phosphorylase a did not effect the rate of dephosphorylation of inhibitor-1 even when it was present in a 50-fold molar excess over inhibitor-1. The result of these three properties is that inhibitor-1 is preferentially dephosphorylated by protein phosphatase-III even in the presence of a large excess of other phosphoprotein substrates. Inhibitor-1 was also dephosphorylated by protein phosphatase-II. The kinetic constants for the dephosphorylation of inhibitor-1 [Km = 2.8 micron, V (rel) = 200] and the alpha-subunit of phosphorylase kinase [Km = 3.7 micron, V (rel) = 100]were comparable...


Occurrence of two fucosyltransferase activities at the outer surface of rat lymphocytes. To demonstrate the existence of ectofucosyltransferase activities on the outer surface of rat lymphocytes, we measured fucosyltransferase activities on whole cells using procedures enabling us to exclude the possibility of misleading results due to precursor hydrolysis and intracellular utilization of the free fucose, and to take into account the contamination by intracellular enzymes freed by the small percentage of broken cells. The described ectofucosyltransferases are able to catalyze the transfer of fucosyl residues from GDP-fucose to the endogenous membrane acceptors but the transfer activity towards exogenous acceptors is restricted to low molecular weight compounds. Use of galactose and di-N-acetylchitobiose as exogenous acceptors and concomitant study of the specific inhibition by N-ethylmaleimide enabled us to detect both types of ectofucosyltransferases: a GDP-fucose: galactoside ectofucosyltransferase and a GDP-fucose: N-acetylglucosaminide ectofucosyltransferase.


Protein kinases of rat liver endoplasmic reticulum. Solubilisation, partial characterisation and comparison with protein kinases of rat liver cytosol. Protein kinase associated with rat liver microsomes was only partly extracted by treatment with 1.5 M KCl. The enzyme was solubilised by Triton X-100 or sodium deoxycholate at the same or slightly higher detergent concentrations than microsomal marker components. The enzyme activity increased 2-3 fold upon solubilisation. Three peaks with protein kinase activity (fractions MI, MII and MIII) were resolved on DEAE-cellulose chromatography. Fraction MIII but not fractions MI or MII was activated by adenosine 3':5'-monophosphate (cyclic AMP). All fractions catalysed the phosphorylation of protamine and histones but not that of casein or phosvitin. Fractions MI and MIII had a similar substrate specificity and phosphorylated histones at a relatively much higher rate than did fraction MII. The isoelectric points were 8.1 for fraction MI, 5.5 for fraction MII and 4.9 for fraction MIII. On incubation of fraction MIII with cyclic AMP it was split into two catalytically active components with pI 8.1 and 7.35. The component with pI 8.1 was predominant and corresponded to fraction MI. Five protein kinase peaks were resolved from rat liver cytosol by DEAE-cellulose chromatography. Three of them (fractions CIa, CIIb and CIII) had the same properties as each of the microsomal kinase fractions.  A forth fraction (CIIa) was cyclic-AMP-dependent and had the same substrate specificity as fractions MI and MIII. Its pI was 5.1, and it was split into two components by cyclic AMP (pI 8.1 and 7.35). In binding studies fraction CIIb bound more efficiently to microsomes than fraction CIII, while fractions CIa, CIIa and the microsomal protein kinase fractions did not bind appreciably. When microsomes were treated with trypsin exposed protein kinase was inactivated and the latency of the remaining enzyme increased substantially. Most of fraction MII was inactivated by trypsin while fraction MIII was resistant. The possible orientation of protein kinase fractions MII and MIII in the microsomal membrane is discussed.


Vascular origin of Poland syndrome? A comparative rheographic study of the vascularisation of the arms in eight patients. Vascularization of the arms has been studied by impedance plethysmography (rheography) in eight children with Poland syndrome, a common malformation characterized by unilateral hand anomaly and ipsilateral aplasia of the inferior head of the pectoralis major muscle. A marked decrease of the velocity of the systolic increase in the arterial volume (Velm) was shown in the affected arms, and the difference between the two arms was significantly higher in the patients than in a control group (P less than 0.02). These results, which are also observed in stenotic atherosclerosis, support the hypothesis of hypoplasia of the ipsilateral subclavian artery as the origin of the malformation, although a local anomaly in arterial-wall viscosity cannot be ruled out. The opacification of the aortic arch in another patient confirmed this hypothesis. Aortography in further patients will be necessary to substantiate our results. However, this technique is not without danger and should only be used during the first months of life when a vascular operation can be envisaged.


The effect of cyproterone acetate on adrenal cortical function in children with precocious puberty. 8 children with precocious puberty were treated with cyproterone acetate (CPA). During treatment there were no definite clinical signs of depressed adrenocortical function. The plasma cortisol concentrations were grossly depressed and the diurnal cortisol rhythm was abolished. Two months after discontinuation of CPA treatment the adrenocortical function had greatly improved. The lysin-vasopressin stimulation test revealed in one child a normal, in another child an exaggerated ACTH response during CPA therapy. Fasting plasma ACTH concentrations were elevated compared with normal controls, but they were very low compared with patients with Addison's disease. The results suggest that CPA has a twofold effect leading to adrenocortical insufficiency: i.e., inhibition of cortisol secretion by the adrenals themselves and inhibition of ACTH secretion at the hypothalamopitiuitary level.


Chronic Niemann-Pick disease with sphingomyelinase deficiency in two brothers with mental retardation. Clinical, biochemical, and electron microscopic studies are presented in two brothers with Niemann-Pick disease. The clinical features include hepatosplenomegaly and mental retardation without any other neurological signs. Roentgenograms of the chest showed bilateral diffuse reticular infiltration. The amounts of sphingomyelin and cholesterol in liver were increased, and sphingomyelinase activities in both liver and skin fibroblasts were markedly reduced in Case 1. Numerous foam cells and myelin figures were observed in the liver, kidneys, bone marrow, and lymph nodes on electron microscopical examination. These cases were regarded as a variant of Niemann-Pick disease from our investigations as they have mental retardation as an exceptional symptom when they are diagnosed as type B.


Free radical intermediates produced by autoxidation of 1,8-dihydroxy-9-anthrone (dithranol) in pyridine. The autoxidation of the antipsoriatic agent dithranol, monitored by an ESR-spectrometer, proceeds through several free radical intermediates. The initial radical, attacking a bulky spin trapping agent in a sterically comparatively hindered constellation, may be the active therapeutic form of dithranol.


Electronmicroscopic identification of type C particles in cultured murine neuroblastoma. Monolayer of murine neuroblastoma were treated with dexamethasone and examined by electronmicroscopy. Most of the treated cells were morphologically differentiated and exhibited type C virus particles which were budding from the cell surface. This in vitro system may be of great value for exploring the oncogenic potential of the virus, and its possible role in cell differentiation.


[The effect of curare-like agents on synaptic transmission in the spinal cord]. In experiments with decerebrated cats subject to study was the influence of new curare-like agents of non-depolarized type of action, such as diadonium, decadonium, RGH-1106, pancuronium-bromide (pavulon) and also of paramion and mellictine on the magnitude of mono-and polysynaptic potentials of the spinal cord, the time of synaptic delay, the lability of the spinal centres and cycle of the motoneurons excitability restoration. Diadonium, decadonium, RGH-1106 and pavulon when used in 1--5-fold myoparalytic doses did not affect the bioelectric activity of the spinal cord. Mellictinum (3,5--4,5 mg/kg)) produced a rise of the monosynaptic responses amplitude by 21 per cent through inhibition of the Renshow n-cholinoreceptors cells. Paramion (0.3--1 mg/kg) suppressed monosynaptic potentials, increasing the afferent proprioceptic activity.


[Possible concentration-dependent mechanism of action exerted by epinephrine on giant neurons]. The results of an investigation carried out suggest that both the stimulating and inhibitory action of epinephrine on the neuronal activity are mediated by the adenylate-cyclase system. The reason of the effect of epinephrine depending on its concentration is, apparantly, the polyfunctionality of the "second mediator"--cAMP and the primordial existence in the cells of processes oppositely influencing its functional activity, whose intensity depends on the concentration of cAMP.


Inhibitory effect of a luteinizing hormone (LH)-releasing hormone agonist on rat ovarian LH and follicle-stimulating hormone receptor levels during pregnancy. When injected at the daily dose of 100 microgram on days 7 through 12 of pregnancy, the luteinizing hormone (LH)-releasing hormone (LHRH) agonist [D-Ala6, des-Gly-NH2(10)] LHRH ethylamide led to complete suppression of pregnancy and a 45% to 60% decrease in plasma progesterone concentration. The antifertility effect of the LHRH analog was accompanied by an early and almost complete inhibition of ovarian LH/human chorionic gonadotropin and follicle-stimulating hormone receptor levels. These data suggest that the antifertility effects of LHRH agonists are mediated by down-regulation of ovarian gonadotropin receptors and decreased luteal function.


Study of group-specific proteins in mammalian cells transformed by Rous sarcoma virus. Lysates of various mammalian RSV-transformed cells labelled with 35S-methionine were tested for the presence of gs proteins and for the size of these molecules by means of antisera, using sodium dodecyl sulphate--polyacrylamide slab gel electrophoresis. In no case was the presence of gs proteins detected in the analyzed immunoprecipitates; only in RSH-mix cells a quantitative difference in the contrast of a 70 000-molecular weight protein was found. The autoradiograms of simultaneously tested immunoprecipitates of lysates of RSV-transformed chicken cells clearly showed the presence of p27 and after a short-term labelling also of the gs precursor molecule (pr76). The cause of the failure in detecting gs proteins by the used method in two mammalian RSV-transformed lines, in which the gs antigens were previously detected by complement fixation test, are discussed.


The use of cytochalasins in studies on the molecular biology of virus--host cell interactions. In conclusion, one can say that the cytochalasins--in their brief history of application in virology--have proven to be valuable tools in studies on the molecular biology of virus--host cell interactions. On the other hand, viral systems can be useful in defining the primary sites of action of cytochalasins in certain cells. In interpreting the effects of cytochalasins on virus replication, however, one must take into consideration that the cytochalasins exert a wide variety of alterations in cellular functions. Infections by viruses interfere primarily with the synthesis of host DNA, RNA, and proteins. The experiments reviewed here were carried out with different viruses, different cell lines, and--more important--under very different experimental conditions. Nevertheless, a number of informative observations emerge which provide insight into specific aspects of the interaction of viruses with their host cells, and help us to understand the mode of action of cytochalasins on specific cellular functions: Cytochalasins may decrease or increase the competence of cells for infection by viruses. For the DNA viruses, vaccinia virus and adenovirus, cytochalasin treatment of host cells resulted in a reduced virus yield; whereas for the RNA viruses, poliovirus, parainfluenza virus, and VSV, cytochalasin treatment results in an increased virus yield. The precise mechanism by which cytochalasins exert these effects remain unclear. It is proposed that the cytochalasin-mediated enhancement of cell infectibility for poliovirus--the only system for which this effect has been studied in more detail--is primarily due to a reduction of polypeptide chain initiation. This reduction dramatically amplifies the inherent translational advantage of virus mRNAs over host cell mRNAs, resulting in a relative stimulation of viral mRNA translation. The cytochalasin-mediated sensitization of cells for infection by isolated poliovirus RNA is explainable by a comparable effect on protein synthesis. In this case, cytochalasin may act in part by substituting for the function of a viral protein(s). When cytochalasin B is added to cells which have previously been infected by intact HSV, formation of infectious virus particles is severely inhibited...


The effect of different regimens of oestrogens on the clotting and fibrinolytic system of the post-menopausal woman. The effects of six different regimens of hormone replacement therapy on coagulation, fibrinolysis and platelet aggregation were sudied in 33 symptomatic, post-menopausal women. Studies were performed before and after 3 months of therapy in the six treatment groups, Premarin, Progynova, Harmogen, Serial 28, ethynodiol diacetate and a testosterone and oestradiol implant. No changes in any parameter were detected following administration of Premarin, Progynova or an implant of testosterone and oestradiol. Serial 28 produced a significant increase in plasminogen concentration and increased the extent of platelet aggregation in 0.5 micron adrenalin solution. Harmogen produced an increase in the extent of platelet aggregation in 1.0 micron adrenalin solution, but a decrease in the rate of platelet aggregation in 1.0 micron solution. Ethynodiol diacetate caused a significant decrease in the kaolin-cephalin clotting time but decreased the extent of platelet aggregation in 0.5 and 1.0 micron solutions of adrenalin. FDPs developed following therapy in 9 of the 33 patients. These were evenly distributed between the groups. No patients had any change in anti-Xa concentration, platelet count or platelet aggregation in thrombin.


[A clinical study on the effect of extrinsic corticotropin on the pituitary-adrenocortical function (author's transl)]. The effects of ACTH preparations on the pituitary-adrenocortical function remain to be clarified in more respects than those of steroid drugs. In the present study, the author investigated the effect of synthetic ACTH-Z on the pituitary-adrenocortical function in a series of 76 patients under synthetic ACTH-Z therapy using the daily urinary excretion of 17-KGS as an index. The results are summarized as follows: 1) The administration of synthetic ACTH-Z in a dose of 0.25 mg or 0.5 mg q.d. was followed by a gradually declining tendency of the daily urinary excretion of 17-KGS. More particularly, after 1.5 months of synthetic ACTH-Z medication at either of these dosage levels, the response of urinary 17-KGS was found to have decreased to about 1/2 of its level at the beginning of medication. 2) The responsiveness of urinary 17-KGS was well maintained even after 3 months of intermittent doses of 0.25 mg of synthetic ACTH-Z (3 times a week, or every other day). 3) Synthetic ACTH-Z, when used in a smaller dose of 0.125 mg daily, produced a minimal stimulating effect on the adrenal cortex. When administered in a less frequent (twice weekly) dose of 0.25 mg, synthetic ACTH-Z failed to prevent or counterbalance atrophy of the adrenal cortex induced by small doses of steroid drugs (average total dose equivalent to 12.2 mg of prednisolone). 4) With the purpose of activating the steroid-atrophied adrenal cortex, synthetic ACTH-Z was administered in a daily or intermittent (every other day, or 3 times a week) dose of 0.25 mg. With the daily dosage regimen, the atrophied adrenal cortex temporarily exhibited a good response, but its responsiveness soon decreased again, and in the end, synthetic ACTH-Z administered in this mode failed to activate the adrenal cortex. With the intermittent dosage regimen, on the other hand, a tendency toward recovery of adrenocortical responsiveness was observed in half the cases studied. In the remaining half receiving steroids over a prolonged period of time, recovery of adrenocortical responsiveness was not attained to any appreciable extent. These results might be interpreted as suggesting that the administration of synthetic ACTH-Z in an intermittent dose of 0.25 mg or 0.5 mg is advisable for the activation of the atrophied adrenal cortex. If this therapeutic regimen fails to initiate a tendency to recovery of adrenocortical responsiveness in a matter of a month or two, this implies that there is atrophy of the adrenal cortex severe enough to invalidate the therapeutic use of synthetic ACTH-Z. 5) The response of the adrenal cortex of 0.25 mg doses of synthetic ACTH-Z was distinctly suppressed by the concomitant use of steroids in 10 mg doses (as calculated on a prednisolone basis). Likewise, the response of the adrenal cortex of 0.5 mg doses of synthetic ACTH-Z was suppressed to 78%, 46% and 16%, respectively, by the conjoined use of steroids in doses of 20 mg, 30 mg and 40 mg or above...


Silica toxicity for macrophages in vitro: a new cytotoxicity test. Soluble factors released from silica-damaged macrophages inhibit proliferation of various haematopoietic cells in long term cultures. The same holds true for damage induced by heat, non-physiological pH, freezing and thawing. This phenomenon is dose-dependent and correlates with the degree of macrophage viability. Thus, a base for measuring the amount of damage to the macrophage is established.


Glucocorticoid modulation of adrenocorticotropin-induced melanogenesis in the Cloudman S-91 melanoma in vitro. The acute in vitro action of adrenocorticotropin (ACTH) and corticosterone alone and in combination were determined in the Cloudman S-91 melanoma grown in vivo. Hormone-treated melanoma dice (5-240 min) were analyzed for tyrosinase activity (EC 1.14.18.1), cyclic AMP (cAMP) and cyclic GMP (cGMP). ACTH elevated cAMP levels in the S-91 melanoma. However, these increases in cAMP were not accompanied by increased tyrosinase activity. Corticosterone depressed cAMP levels while stimulating tyrosinase activity. ACTH plus corticosterone produced an early cAMP peak followed by depression. ACTH plus corticosterone stimulated tyrosine activity coincident with the early cAMP peak followed by a drop in tyrosinase activity which was subsequently elevated. cGMP levels were not altered by any hormone treatment. The results indicate that cAMP is not the sole modulator of tyrosinase activity and suggest the interaction of ACTH, corticosterone and cAMP in the regulation of melanoma tyrosinase activity.


Speech, language and hearing in Moebius syndrome: a study of 22 patients. A comprehensive profile of communicative disorders in patients with Moebius syndrome was attempted. Seven patients were evaluated by the authors and the findings were added to 15 contributed case-histories. Most patients demonstrated some degree of dysarthria due to congenital paralysis of the facial nerves. Other problems such as cleft palate, hearing loss, mental retardation and delayed development of language were found in a few of the patients. As most of the older children in the sample and in reports in the literature eventually developed intelligible speech, a program of oral language stimulation and compensatory articulatory adjustments would appear to be the procedure of choice.


The effect of induced hyperglucagonaemia on the Zucker fatty rat. The effect of chronic treatment with a long-acting glucagon preparation on liver glucagon and insulin receptors, adenylate cyclase and plasma lipids has been examined in Zucker fatty rats (fa/fa) and their lean littermates (Fa/-). Liver insulin and glucagon receptors were examined using radioreceptor assay techniques. Neither fatty nor lean rats showed any change in insulin receptors after glucagon treatment. Glucagon receptors of the fatty rats showed a 33% drop in the number of the glucagon receptors after glucagon treatment, whilst there was no such change in the lean group. Plasma membranes of the treated fatty rats and their controls bound only 50% as much insulin per mg of liver membrane protein as those of the treated lean rats and their controls. Glucagon treatment raised plasma NEFA in lean rats and reduced them in fatty ones. Plasma cholesterol levels were reduced in both groups of animals as were plasma triglycerides, though to a lesser degree in fatty than in lean animals. Glucagon treatment increased basal and stimulated adenylate cyclase activity in the lean rats and even more so in the fatty ones. The data lend no support to the concept that hypertriglyceridaemia in fatty Zucker rats is a consequence of abnormal glucagon responsiveness.


Experimental chemotherapy of carcinoma of the human stomach and colon serially transplanted in nude mice. Human gastric and colon carcinomas serially transplantable in nude mice, designated as St-4, St-15, and Co-3, were used for chemotherapeutic experiments. All tumors showed a 100% take rate and stable growth. Mitomycin-C (MMC), 3 mg/kg body weight, given intraperitoneally 4 times once weekly, showed some inhibiting effect on Co-3, St-15, and St-4 in that order. N1-(2'-Tetrahydrofuryl)-5-fluorouracil (FT-207), 90 mg/kg body weight, intraperitoneally daily for 4 weeks, suppressed the growth of Co-3 and St-15 slightly but with statistical significance, and had no effect on St-4. Co-3, a tumor with more rapid growth and richer vascular supply, was suppressed more by MMC and FT-207. These three transplantable tumors may be useful, not only in evaluating chemotherapeutic agents for use against human gastrointestinal carcinomas, but also for screening of new antitumor agents.


Enhancing and inhibitory effects of some stilbene and steroid compounds on induction of hepatoma in rats fed 3'-methyl-4-(dimethylamino)azobenzene. Examinations were made on substances that enhance or inhibit the induction of hepatoma in rats previously fed 3'-methyl-4-(dimethylamino)azobenzene (3'-Me-DAB) for a brief period. The substances tested were stilbene, 4-nitrostilbene, 4,4'-dihydroxystilbene, diethylstilbestrol, 17beta-estradiol, and methyltestosterone. Male Donryu rats were fed 0.5 g of 3'-Me-DAB by being maintained on a diet containing 0.06% 3'-Me-DAB, and then they were fed 0.25 or 0.5 g of a test substance with the basal diet. Comparison of the development and yield of hepatomas indicated that 4-nitrostilbene and methyltestosterone had an activity of enhancing 3'-Me-DAB carcinogenesis, whereas diethylstilbestrol and 17beta-estradiol had an activity to retard it. Other substances showed no such activities. The enhancement by 4-nitrostilbene and inhibition by diethylstilbestrol of 3'-Me-DAB carcinogenesis was correlated with their effect on liver nucleic acid metabolism. Feeding of 4-nitrostilbene caused a selective inhibition of Mn2+-(NH4)2SO4-activated RNA polymerase activity of liver nuclei and reduced liver RNA content. The deleterious alteration of liver RNA metabolism was followed by the enhancement in the incorporation of ip-injected 3H-thymidine into DNA of liver nuclei. On the other hand, feeding of diethylstilbestrol increased tissue RNA content without effect on RNA polymerase activity of liver nuclei, and had an activity of increasing the incorporation of 3H-thymidine into DNA. The possible implication of these results with regard to the enhancement and inhibition of hepatocarcinogenesis is discussed.


Transformation of Indian muntjac cells by murine and avian sarcoma viruses. Indian muntjac cells were efficiently transformed by murine sarcoma virus (MSV) and avian sarcoma viruses (ASV). When colony formation of the infected cells was examined in soft agar, many colonies were formed by the ASV-injected cells but no colony was seen in the MSV-infected cells. The ASV-transformed cell clones differed among the clones in morphology, presence of inducible ASV genome, and karyotypes.


Granular cell tumors of the esophagus.

Granular cell tumors of the esophagus. Three cases of granular cell tumor of the esophagus are added to the 17 previously reported in the literature. These tumors, thought to be of neural origin, are difficult to diagnose preoperatively. The diagnosis should be considered in adult females presenting with an intramural mass of the proximal or distal third of the esophagus. Symptoms of dysphagia and substernal discomfort are likely to occur with lesions greater than one centimeter in diameter. Preoperative biopsy is not advised as a mistaken diagnosis of squamous cell carcinoma can result.


Calcified primary tumors of the gastrointestinal tract. The dominant pattern and location of calcifications occurring within 23 primary gastrointestinal tumors have been analysed and correlated with the data from the literature. The provided guidelines for radiologic diagnosis of such calcified tumors include: (1) a retrocardiac mass containing amorphous calcifications is typical of leiomyoma of the esophagus; (2) calcific deposits similar to that in uterine fibroids may be the feature of gastric leiomyoma or intestinal leiomyosarcoma; (3) sand-like deposits within the wall of the stomach or colon are characteristic of a mucinous adenocarcinoma; (4) clusters of phleboliths in the gastrointestinal wall suggest a hemangioma particularly if recurrent intestinal bleeding and cutaneous hemangiomas are associated; (5) sunburst type of calcification in the pancreas indicates a cystadenoma or cystadenocarcinoma of that organ; and (6) aggregates of granular calcifications in the liver are diagnostic for metastatic adenocarcinoma of the colon but may rarely be seen in a primary malignancy of the liver.


Duodeno-duodenal fistula--a manifestation of carcinoma of the colon. Two cases of duodeno-duodenal fistula complicating carcinoma of the right colon, both presenting with upper gastrointestinal hemorrhage, are presented. The close proximity of the ascending colon and hepatic flexure and their draining lymphatics to the duodenum are discussed.


A case of rectal carcinoma concomitant with Pagetoid lesion in the perianal region--histopathological and electron microscopic observations--. A case of extramammary Paget's disease of the perianal area adjacent to a mucinous adenocarcinoma of the rectum is presented. The histopathological examination of the case revealed that within the epidermis adjacent to the invading rectal cancer nest, the formation of glandular structures can be observed occasionally, around which so-called Paget's cells are noted to be scattered and further many points of similarity between cancer cell and Paget's cell are proved by electron microscope. Perianal skin lesion looking like Paget's disease is proved to be due to intraepidermal spread of the rectal carcinoma. Referring to the literature, this is the first report to our knowledge, to elucidate the histogenesis of perianal extramammary Paget's disease associated with a rectal carcinoma using the electron microscopy.


Lipoprotein-X levels in extrahepatic versus intrahepatic cholestasis. Lipoprotein-X (LP-X) is an abnormal lipoprotein characteristic of cholestasis. To assess recent claims that its serum concentration helps differentiate extrahepatic from intrahepatic cholestasis, we studied 42 consecutive LP-X-positive patients. The mean serum LP-X level was higher in 18 patients with extrahepatic obstruction than in 24 with intrahepatic cholestatis, 321 +/- 89 versus 130 +/- 31 SEM mg per dl (P less than 0.05). However, values overlapped in 38 of the 42 cases, and in the other 4 the diagnosis of extrahepatic obstruction was obvious anyway from clinical examination. LP-X concentration gave little more information than did free cholesterol of phospholipid levels, which it closely paralleled (r = 0.89 and 0.81, respectively, P less than 0.01). There was no correlation with standard liver function tests or with activity of lecithin-cholesterol acyltransferase, the enzyme responsible for cholesterol esterification in plasma. Contrary to recent claims, these findings suggest that serum LP-X quantitation has little or no clinical value in distinguishing extrahepatic from intrahepatic cholestasis.


[Diagnosis of tumor or metastasis in the liver by means of laparoscopy and gammagraphy]. Fifty-four patients with proven malignancy of the gastrointestinal tract or suspicion of primary or metastatic hepatic carcinoma were studied to evaluate the effectiveness of different diagnostic methods. Gammagraphic and laparoscopic evaluations were emphasized. The results indicated that laparoscopy was the most effective method for diagnosing primary or metastatic neoplastic liver disease.


Stimulation by gastro-intestinal hormones of cyclic adenosine 3': 5'-monophosphate accumulation and insulin release in isolated pancreatic islets of the rat. In collagenase isolated rat pancreatic islets, CCK-PZ, SHG, secretin and glucagon stimulated the accumulation of cAMP, in physiological ranges. The resulting increment of cAMP showed a good correlation with insulin release stimulated by either glucagon or secretin, but not by SHG or CCK-PZ. In the same system, 14CO2 production from glucose-U-14C was significantly increased by either SHG or CCK-PZ. The results presented in this report are compatible with the hypothesis that insulin release by gastrointestinal hormones may be mediated by cAMP in the B-cell in the case of either glucagon or secretin; whereas, in the case of either SHG or CCK-PZ, it may presumably be mediated by an unknown mechanism in glucose metabolism, other than c-AMP.


[Angiographic aspects of tumors of the islet of the pancreas]. Four patients with islet cell tumours were explored with selective angiography. The radiological findings of 3 insulinomas and 1 nonfunctioning tumor are described. The typical hypervascular pattern of parenchimatose phase was found in all. The three insulinomas were successfully treated surgically. The nonfunctioning tumor was not removed due to the pathologists original impression that it was a maligant tumor involving the portal vein. Angiographic pattern, etiology, differential diagnosis and causes of errors are discussed. The method is diagnostically useful and offers important precperative information thus reducing operating time and minimizing the magnitude of the surgery.


Canine kidney cells: I. Neutral lipids, phospholipids and fatty acids of SV40 transformed canine kidney cells in suspension and monolayer culture. Differences in lipid composition and the effects of monolayer and suspension culture on SV40 virus transformed canine kidney cells and their plasma membranes were studied. Plasma membranes from monolayer and suspension cultures of SV40 transformed canine kidney cells were isolated by the Warren fluorescein-mercuric acetate technique. Free cholesterol was the major neutral lipid extracted from the whole cells and plasma membranes of all these cultures. Triglyceride increased about 6- to 50-fold in suspension grown whole cells and their plasma membranes compared to those from monolayer cultures. The mu moles of total neutral lipids and phospholipids are elevated in plasma membranes compared to whole cells from monolayer cultures but are similar in suspension cultures. The distribution of fatty acids in the major lipid classes of plasma membranes and whole cells generally follow the composition of calf serum used in the culture media. INDEX WORDS: SV40, plasma membranes, transformed cells, canine kidney cells, neutral lipids, phospholipids, monolayer culture, suspension culture.


[Effects of selective applications of drugs affecting ganglionic transmission to the hypogastric ganglion of the guinea pig (author's transl)]. Effects of selective applications of drugs to the guinea pig hypogastric ganglion were studied on the contractile response of the vas deferens to the hypogastric nerve stimulation or to ganglionic stimulating drugs. Both acetylcholine (ACh) and dimethylphenylpiperazinium (DMPP) contracted the vas deferens. Neostigmine  potentiated the ACh-induced contractions but not those which were DMPP-induced. Hemicholinium-3 (HC-3) reduced the response to nerve stimulation (R-NS) but affected neither the ACh-induced contractions nor the DMPP-induced contractions. Morphine blocked R-NS but not the ACh-induced and the DMPP-induced contractions. Hexamethonium reduced R-NS and the contractions to ACh or to DMPP. Methacholine and bethanechol applied before and after preganglionic tetanic stimulation of the hypogastric nerve produced no change of tone of the vas deferens. Atropine did not antagonize the responses which were produced by nerve stimulation, ACh or DMPP. These results support the classic hypothesis that ganglionic transmission is mediated by nicotinic receptors but do not provide evidence that muscarinic receptors exist at the synapses of this preparation.


[The medical after care of patients with stomas]. The medical care of the increasing number of ostomy patients is necessary in three inseparable areas:--provision of the stoma,--treatment of basis disease,--psychological support. The major medical and technical advances of recent years allow a normal social life. A precisely defined program of examinations at regular intervals is necessary to ensure adequate treatment; especially after Ca-operations. Psychological support must be directed towards the prevention of social isolation. In order to improve the present situation in the German Federal Republic the graduate training of the doctors must be intensified. In order to deal with problematical cases special ostomy out-patient clinics with suitably trained personnel should be set up in certain hospitals.


Glycogen metabolism in the placenta of streptozotocin diabetic rats. Administration of STZ prior to mating, induced significant impairment in glycogen metabolism in term rat placentae. Glycogen retention was observed in the treated placentae, while glycogen synthetase activity was reduced. The glycogenolytic pathway seemed to proceed mainly through amyloglucosidase enzyme whose activity increased threefold. Glucose-6-phosphatase showed a moderate activation, mainly in the labyrinth zone, while phosphorylase was slightly inhibited. These changes were accompanied by a decrease in placental insulin levels. The possible defensive role played by the placenta in cases of maternal hyperglycemia was postulated.


Effect of epinephrine and insulin on adenosine 3'5'-cyclic monophosphate--dependent protein kinase in human skeletal muscle in vivo. Cyclic AMP dependent protein kinase has beeen identified in human skeletal muscle tissue. In crude muscle extracts the enzyme was 3--5 fold activated by cyclic AMP. The cyclic AMP-dependent activity (corresponding to the inactive holoenzyme) was completely inhibited by the heat stable inhibitor of protein kinase. Reciprocal changes of the cyclic AMP-dependent activity in skeletal muscle were observed after administration of epinephrine and insulin in vivo. Infusion of epinephrine in healthy volunteers increased the level of cyclic AMP and decreased the activity of the cyclic AMP-depenent form (i.e. the inactive form) of protein kinase. These changes were reversible after cessation of epinephrine administration. The results are consistent with an activation of protein kinase in vivo due to an epinephrine mediated increase of the concentration of cyclic AMP. I.v. injection of insulin had the opposite effect on the enzyme in skeletal muscle, leading to increased activity of the cyclic AMP-dependent form of protein kinase. Insulin had no effect on the level of cyclic AMP, but promoted a transient increase of cyclic GMP 1 min. after insulin injection. The effect by insulin on protein kinase cannot be related to the level of cyclic AMP or cyclic GMP.


Effect of acute ethanol load on plasma immunoreactive insulin and glucagon. The effect of a single large dose of ethanol (5 mg/kg body weight) on plasma glucagon (IRG) and insulin (IRI) concentrations was studied in rats fasting for 24 hr. Hepatic cAMP concentration and blood glucose were also estimated and correlated with hormonal changes. Plasma IRG concentrations had doubled by the first sampling time (2 hr) and remained at this level up to 16 hr after ethanol administration. Plasma IRI concentrations were not affected by ethanol. Hepatic cAMP concentrations reflected changes in the plasma insulin/glucagon ratio, which seems to be the major determining factor for hepatic cAMP even during ethanol oxidation. Hypoglycemia was not found in the ethanol group during the experimental period of 24 hr, and it was therefore concluded that ethanol may stimulate glucagon secretion in rats even without concurrent hypoglycemia. Possible mechanisms for the action of ethanol on the endocrine pancrease are discussed.


Influence of genetic obesity in mice on the lipolytic response of isolated adipocytes to isoproterenol and ACTH-(1-24). The lipolytic response of isolated adipocytes from genetic obese (C57/BL/64 ob/ob) and lean (C57BL/6J +/?) mice to ACTH-(1-24), isoproterenol and glucagon has been studied. The mean cell idameter of adipocytes form ob/ob mice was approximately twice that of lean controls. The adipocytes from obese mice contained on the average approximately six times the amount of triacylglycerol present in the smaller lean mouse adipocyte. Lipolysis was calculated both on a per cell basis (10(5) cells) and per mu mole of triacylglycerol and when expressed on a cell number basis, the larger adipocytes from obese mice showed an ACTH-(1-24) stimulated glycerol release which was quantitatively similar to that of smaller adipocytes from lean mice. When expressed per mu mole of triacylglycerol, the smaller cells from lean animals appeared to be dramatically more responsive to either isoproterenol or ACTH-(1-24). On either basis, ACTH-(1-24) stimulated glycerol release from obese mouse cells was greater than the isoproterenol response. The obese mouse of adipocyte showed selective loss of response to isoproterenol compared to its lean control.


An increase of pituitary 3', 5' cyclic adenosine monophosphate produced by estradiol benzoate in vitro: possible implication of this increase in the secretion of luteinizing hormone. In an attempt to study the site and mechanism of action of estrogen in producing positive feedback control, porcine anterior pituitary slices were incubated in vitro in the presence of estradiol benzoate (EB). EB elevated pituitary cyclic AMP concentration within 5 min and augmented pituitary release of luteinizing hormone (LH). The magnitude of increase of cyclic AMP and LH release was related to the doses of EB used. Also, luteinizing hormone releasing hormone (LH-RH) elevated pituitary cyclic AMP concentration and stimulated pituitary release of LH. The magnitude of increase of cyclic AMP and LH release was inversely related to the doses of LH-RH used. EB and LH-RH were additive in increasing cyclic AMP. Progesterone and clomiphene citrate interfered with an increase of pituitary cyclic AMP produced by EB, but did not significantly affect the basal level of pituitary cyclic AMP. Testosterone propionate, human chorionic gonadotropin and hexestrol were without effect on either basal or stimulated level of pituitary cyclic AMP. Since cyclic AMP and dibutyryl cyclic AMP (DBC) stimulated LH release, it is suggested that EB directly stimulates the release of LH by augmenting cyclic AMP synthesis in the anterior pituitary.


The effect of cyclic nucleotides on the incorporation of 3H-glucosamine into hyaluronate in bone organ culture. Addition of dibutyryl cyclic AMP or parathyroid hormone to bone organ cultures markedly increased the incorporation of 3H-glucosamine into non-dialyzable macromolecules. Other cyclic nucleotides or their dibutyryl derivatives did not stimulate glucosamine incorporation. DEAE-cellulose chromatography of the papain-digested calvaria and culture medium resolved the labeled material into four peaks. A four-fold increase in radioactivity was observed in peak III. Previous studies of peak III have identified the labeled material as hyaluronic acid. The results suggest that the parathyroid hormone stimulated increase in glucosamine incorporation is mediated via the adenylate cyclase-cyclic AMP system, and that the increased amount of radioactivity is due to an increased amount of hyaluronic acid. Turnover studied of the labeled material suggest that the release of proteoglycans into the culture medium is not inhibited in the cultures treated with dibutyryl cyclic AMP. The role of hyaluronate in this experimental system remains to be elucidated.


Changes in the microheterogeneity of histone H1 after mengovirus infection of Ehrlich ascites tumor cells. Employing high-resolution polyacryl-amide gradient slab-gel electrophoresis in 6M urea and 6% acetic acid, a distinct change in the microheterogeneity of histone H1 was detected after mengovirus infection of Ehrlich ascites tumor cells. Whereas in uninfected cells the band multiplicity was found to be 6, it was reduced to 4 in the course of the infectious cycle (8 to 9 h). It could be deomonstrated that, while the electrophoretically slowly moving subspecies H1e and H1f disappeared from the band profile of histone H1, the faster migrating bands H1a and H1b increased in relative intensity. The relative intensity of band H1d was also drastically reduced, that of band H1c stayed practically constant throughout infection. When Ehrlich ascites tumor cells were labeled with a mixture of [14C]amino acids prior to mengovirus infection, the radioactivity incorporated into histone H1 subspecies of low electrophoretic mobility was chased into histone H1 components of high mobility in the course of infection, suggesting a virus-induced, unidirectional interconversion of the multiple histone H1 subunits. With the exception of a histone H2B subspecies, which also decreased in amount, the relative quantities of the other histones stayed constant throughout infection.


Enhanced phosphorylation of ribosomal protein s6 and other cytoplasmic proteins after mengovirus infection of Ehrlich ascites tumor cells. After mengovirus infection of Ehrlich ascites tumor cells, an increased incorporation of radioactive phosphate into ribosomal protein S6 was detected. As judged from the shape of the radioautography spot and the location of the modified protein on the two-dimensional polyacrylamide gel, it must be concluded that more than one phosphate group was incorporated into ribosomal protein S6. Concomitantly, multiple cytoplasmic proteins, particularly those with high affinity for ribosomes, were phosphorylated. Compared with the controls (mock infection), mengovirus infection induced only a slight additional phosphorylation of the proteins of the ribosome-free supernatant. Employing the radio isotope dilution test with 3', 5'-cyclic AMP-binding protein, no enhancement of the intracellular concentration of 3', 5'-cyclic AMP in response to mengovirus infection could be deomonstrated.


Multiple protein kinases from Trypanosoma gambiense. Three protein kinases were distinguished in Trypanosoma gambiense extract. The enzymes preferred phosvitin, histone, and protamine as acceptor proteins, respectively. The amino acid residues of the acceptor proteins which were phosphorylated by these protein-kinase activities were serine and- to less extent- threonine. The protein kinase activities were neither affected by cyclic nucleotides nor by cyclic AMP receptors. The molecular weights of these protein kinases were determined to be greater than 200,000, 95000 and 37000, respectively. The activities of all three protein kinases were affected to varying degrees by nucleotides and nucleosides.


Adenosine 3', 5'-cyclic monophosphate-binding proteins from Trypanosoma gambiense. Two cyclic AMP-binding proteins, not identical with regulatory subunits of protein kinases, have been isolated from Trypanosoma gambiense. The cyclic AMP receptors were separated by gel chromatography on the basis of their molecular weights. The binding constants of the high and the low molecular weight receptors for cyclic AMP were determined to be 0.4 muM and 0.6 muM, respectively. Cyclic IMP and cyclic GMP compete with cyclic AMP for the binding sites of both receptors. The cyclic AMP binding of the low molecular weight receptor was competitively inhibitied by adenine derivatives. The binding capacity of the high molecular weight receptor was enhanced about two-fold by proteolytic modification with trypsin.


Studies on the biosynthesis of cyclitols, XXXVI. Purification of myo-inositol-1-phosphate synthase of the duckweed, Lemna gibba, to homogeneity by affinity chromatography on NAD-Sepharose molecular and catalytic properties of the enzyme. Using the technique of affinity chromatography on NAD-Sepharose the myo-inositol-1-phosphate synthase of Lemna gibba was purified to homogeneity. The molecular and catalytic properties of this enzyme differ very much from those of myo-inositol-1-phosphate synthase from animal sources. Thus the specific activity of the duckweed enzyme is more than two orders of magnitude higher than that of the enzyme from rat testes. It is inhibitied by EDTA and can be reactivated by Mn2. Its molecular weight (135000 +/- 5000), its subunit composition (3 subunits with identical electrophoretic behaviour) and its isoelectric point (pH 7.7) are also very different from the corresponding parameters for the animal enzyme.


Carcinomas of intraoral salivary glands. Of 44 intraoral salivary gland tumours received by the Department of Pathology, Dental Faculty, University of Oslo over the last 20 years, 21 (approx. 48%) were classified as carcinomas. Of these, the adenocarcinomas constituted the largest group, but because some of them bore a certain likeness to adenoid cystic carcinomas and to mucoepidermoid tumours, we believe that transitions exist between these groups of neoplasms. The material is thought to reflect the daily practice of oral surgeons and general dental practitioners, and we think it important to emphasize that approximately every second tumour in this series was malignant.


Yellow-brown spindle bodies in mesenteric lymph nodes: a possible relationship with melanosis coli. Four cases are described in which spindle-shaped, yellow-brown bodies were seen in the mesenteric lymph nodes of patients with melanosis coli. A comparison of the staining reactions and ultrastructural appearances of the spindle bodies and melanosis pigment suggest that they are related within the broad group of lipofuscins and that the spindle bodies are formed as a result of coalescence of lysosomes containing the pigment.


Carcinoma of the breast with metaplasia to chondrosarcoma: a light and electron microscopic study. Two carcinomas of the breast containing large areas of sarcomatous tissue were studied by light and electron microscopy. In one of these, the sarcomatous element was frankly cartilaginous and in the other, predominantly myxoid but with small cartilaginous-looking foci. By light microscopy, a highly suggestive metaplastic transition could be traced from cells within the epithelial nests to those within the sarcomatous lobules. Ultrastructurally, cells in the former region showed epithelial characteristics and those in the latter region, mesenchymal and/or cartilaginous features. The carcinomatous cells contained desmosomes and formed intercellular spaces lined by microvilli; a few cells showed prominent profiles of rough endoplasmic reticulum. In the first case, the cells in the immediate vicinity of the epithelial nests and those in the fully developed cartilaginous regions showed a progressive dilatation of their endoplasmic reticulum to form large sac-like structures filled with a finely granular and floccular material. The intercellular matrix was electron lucent and contained scattered dense particles, fibrillo-granular material and collagen fibres. Condensation of this material at some distance from the cell resulted in the formation of lacunae. In the second case, the cells in the myxoid areas also showed prominent dilatation of endoplasmic reticulum.


Primary signet ring cell carcinoma of the breast. Three examples of primary signet ring cell carcinoma of the breast are described. This form of breast carcinoma deserves recognition as an entity because its prognosis may differ from colloid carcinoma of the breast and because of potential difficulty in distinguishing it from metastatic carcinoma.