Catecholamine turnover measurement and ACTH-induced short-term changes of catecholamine levels in individual brain nuclei. The effect of acute ACTH treatment on catecholamine content of various brain nuclei was followed by radioenzymatic assay. The hormonal treatment employed affected catecholamine levels only in some hypothalamic nuclei and in the locus coeruleus. Some effects could be detected as rapidly as 10 min after the injection of the hormone. Employing alpha-methyl-para-tyrosine, catecholamine turnover in individual nuclei was estimated, and shown to be quite variable from one area to the other. However, severe limitations seem to exist in vivo, which restrict the feasibility of this approach to basal or chronic situations.


A non-invasive test for receptor binding applied to nephrogenic diabetes insipidus. Studies in animals have determined the importance of specific receptors to the action of many hormones and drugs. In man, a non-invasive external counting technique has been used and absence of receptor function has been demonstrated in a patient with nephrogenic diabetes insipidus using radioactively labelled arginine vasopressin. This is in contrast to the findings in a patient with pituitary diabetes insipidus and a normal control. These results suggest a model for the study of hormone and drug kinetics in man avoiding multiple samplings of biological fluids.


Recent advances in the identification of hepatitis viruses. Resulting directly from the discovery of virus-related antigens, rapid progress has marked the last decade of viral hepatitis research. The hepatitis B virion has been tentatively identified as a DNA virus with an endogenous DNA polymerase, and new serological markers for type B hepatitis have been discovered. Hepatitis A antigen has been identified on a virus-like particle thought to be the hepatitis A virion. Progressively more sophisticated assays for hepatitis antigens and antibodies have been applied to the study of viral hepatitis epidemiology and biochemical-biophysical characterization of the agents. Most recently, knowledge learned from such studies has been exploited to develop a prototype non-infectious but immunogenic hepatitis B vaccine using hepatitis B surface antigen (HBsAg) purified in large quantities from chronic HBsAg carriers. Especially exciting is the prospect, suggested by serological studies of viral hepatitis, that hepatitis viruses besides hepatitis A and B viruses will be identified.


Hepatoma causing a massive tumour embolus. A case of hepatoma is reported, with a typical clinical setting but a most unusual and dramatic final episode. A massive tumour embolus weighing 30 g became arrested on the tricuspid valve, resulting in acute circulatory failure.


[Reference values of cardiac volume in children and adults of the Ivory Coast]. The cardiac volume in decubitus was measured by Rohrer radiological method in 229 subjects, of which 150 were children of both sexes, aged 4 to 16 and 79 were adult males, 17 to 50 years old. Anthropometric parameters and maximum O2 consumption were also measured. Considering that the size of thorax in Ivory Coast natives is significantly smaller than that of a Caucasian, an eventual influence over the cardiac volume was looked for. Results showed that: -- in the child, the cardiac, volume increased linearly with age: 162 ml at 4 and 680 ml at 16; -- significant positive correlations were found between the cardiac volume, the body-weight andusrface, and the measurements of the thorax; -- when compared to data found in the literature, the cardiac volumes of children and adults of the Ivory Coast in relation with the body weight or surface are lower than the values found in Caucasians; -- children who have been swimming for two years and adults from the  national swimming team have a cardiac volume significantly superior to that of Ivory controls; for these subjects, results are equivalent to those of Caucasians; -- there is a highly significant positive correlation between the cardiac volume and the maximum oxygen consumption. To compare values of cardiac volume in different populations, it is necessary that the groups of subjects be very homogeneous and have a comparable level of activity.


Neurophysiological aspects of peripheral neuropathies. 1. Eighty-eight intrafascicular neural recordings were obtained in 10 normal subjects, 5 patients with axonal degeneration and 11 patients with demyelinating neuropathy. 2. Stimulus levels required for perception and fibre activation were higher in neuropathic subjects. Fibres transmitting touch perception had significantly lower conduction velocities in both patient groups, but were very much lower in the group with demyelinating neuropahty than the group with axonal degeneration. Maximum electrical stimulation evoked dispersed fibre responses in the axonal degeneration group and more dispersed, slowly conducting fibre potentials in the demyelinating group. In patients with hypertrophic Charcot-Marie-Tooth disorder, usually only a small group of slowly conducting low amplitude potentials was recorded. 3. Delivery of a train of supramaximal stimuli caused prolongation of latency and dispersion of fibre potentials in all microneurographic recordings. The changes were significantly greater in the axonal neuropathy group than in normals, and recovery was slower. The demyelinating neuropathies showed significantly greater changes than both the normal and the axonal neuropathy groups, and post-tetanic conduction slowing became even more marked after limb temperature was raised. 4. Surface SAP recordings showed normal refractory period in chronic axonal neuropathy but significant latency prolongation occurred in demyelinating neuropathy. 5. It is concluded that both receptor and nerve fibre abnormalities contribute to sensory dysfunction in degenerative and demyelinating neuropathies.


[Sensitivity of thyrotoxic heart to adrenergic influences]. Sensitivity of the cardiovascular system of patients suffering from thyrotoxicosis to exogenous adrenaline was studied. In difference from healthy persons, a single injection of 1 ml of 0.1% adrenaline solution to patients with thyrotoxicosis of moderate severity caused a significant elevation of the arterial pressure, frequency of cardiac contractions, systolic and minute output, myocardiac inotropism indices. An increased inderal reactivity was noted in these patients. Thus, greater sensitivity to adrenergic actions and to beta-adrenoblock was observed in conditions of an excess of thyroid hormones. A coincident administration of adrenaline and inderal balanced the positive chronoinotropic effect of the former, but failed to prevent the hypertensive effect of adrenaline.


[Effect of enteral administration of sodium ribonucleate on the synthesis of amino acyl t RNA in the liver and skeletal muscles of rabbits in experimental hypercorticism]. Daily, for 14 days, rabbits of one group were injected with corticotropin, i.e. ACTH-zinc-phosphate (10 units/kg), whereas rabbits of another group were given (in addition) sodium ribonucleate (40 mg/kg) through a tube into the stomach. Formation of lysyl-tRNA, leucyl-tRNA, and alanyl-tRNA in the liver and the skeletal muscles proved to be significantly greater in the animals which received ACTH together with sodium ribonucleate, as compared to that in the animals given the hormone alone. Hyperglycemia, hepatomegaly, and emaciation were less pronounced in the animals given both the preparations.


Heat capacity and entropy changes in processes involving proteins. Six possible sources of the large heat capacity and entropy changes frequently observed for processes involving proteins are identified. Of these the conformational, hydrophobic, and vibrational effects seem likely to be of greatest importance. A method is proposed for estimating the magnitudes of the hydrophobic and vibrational contributions. Application of this method to several protein processes appears to achieve significant clarification of previously confusing and apparently contradictory data.


Inhibition of DNA synthesis in cultures of 3T3 cells by isolated surface membranes. When added to a sparse culture of 3T3 cells, a surface membrane-enriched fraction from 3T3 cells inhibited the rate of DNA synthesis in a time- and concentration-dependent manner. The membrane preparation had no effect on the rate of DNA synthesis of simian virus 40-transformed 3T3 cells. A similar membrane preparation from transformed cells had a lesser inhibitory effect on 3T3 cells and no effect on transformed cells. The inhibition by membranes was reversible. The data suggest that, when added to growing 3T3 cells, 3T3 surface membranes can mimic the effect of increasing cell density on DNA synthesis.


Characterization of a rat liver factor that inhibits initiation of protein synthesis in rabbit reticulocyte lysates. Protein synthesis in rabbit reticulocytes and their lysates is regulated by heme. In heme-deficient reticulocyte lysates, protein synthesis proceeds at the initial rate for several minutes and then declines abruptly. Inhibition of protein synthesis is due to the activation of a heme-regulated translational inhibitor (HRI) which blocks the initiation of protein synthesis. Addition of the isolated HRI to hemin-supplemented lysates causes inhibition of initiation similar to that observed in heme-deficiency. HRI has been shown to be a protein kinase that specifically phosphorylates the Met-tRNA(f) binding factor (eIF-2). We have isolated an inhibitor (LI) of protein chain initiation from rat liver which displays properties similar to those of HRI: (i) the chromatographic behavior of LI on DEAE-Sephadex, DEAE-cellulose, and phosphocellulose is similar to that of HRI; (ii) both LI and HRI inhibit protein chain initiation in rabbit reticulocyte lysates with the same kinetics of inhibition-i.e., an initial period of synthesis for several minutes at the control rate followed by an abrupt decline in the rate of initiation; (iii) both inhibitions are prevented or reversed by eIF-2; (iv) GTP (2 mM) prevents, and ATP (2 mM) potentiates, the inhibition of protein synthesis induced by either inhibitor; (v) LI is associated with a protein kinase that also phosphorylates the 38,000-dalton subunit of elF-2. These findings indicate that a mechanism for the regulation of protein synthesis similar to that found in rabbit reticulocytes may be present in rat liver.


Isolation of amino acid activating subunit--pantetheine protein complexes: their role in chain elongation in tyrocidine synthesis. Dissociation of the multienzymes of tyrocidine synthesis by prolonged incubation of crude extracts of Bacillus brevis (Dubos strain, ATCC 8185) has yielded, on Sephadex G-100 chromatography, two fractions of amino acid activating subunits, a larger one of 70,000 daltons and a smaller one of 90,000 daltons; the latter was a complex consisting of the 70,000 dalton subunit and the pantetheine-carrying protein of about 20,000 daltons. When it dissociated, the intermediate enzyme, which activates three amino acids, contained two-thirds of the subunits in the 70,000 dalton and one-third in the 90,000 dalton fraction; the heavy enzyme, which activates six amino acids, contained five-sixths of the subunits in the former fraction and one-sixth in the latter. Both fractions showed ATP-PP(i) exchange with all amino acids that are activated by the respective polyenzymes. With proline as an example, the 70,000 dalton subunit exhibited a single low-affinity binding site, which should correspond to the peripheral thiol acceptor site, whereas the 90,000 dalton subunit showed both a low-affinity binding site and an additional high-affinity site for proline; the high-affinity site is attributed to the pantetheine present on the pantetheine-carrying protein, and suggests that amino acids are translocated from the peripheral SH to the pantetheine-carrying moiety during chain elongation. This was confirmed by the observation that the 90,000 dalton complex, when incubated with the light enzyme in the presence of phenylalanine and proline, produced DPhe-Pro dipeptide that cyclized into DPhe-Pro diketopiperazine, but the 70,000 dalton activating subunit, when similarly incubated, did not. After subunit dissociation, however, no further elongation occurred after the transfer from phenylalanine to proline.


3':5'-cyclic AMP-dependent 3':5'-cyclic GMP accumulation in Dictyostelium discoideum. Suspensions of 3':5'-cyclic AMP (cAMP)-sensitive cells of Dictyostelium discoideum responded to a cAMP pulse with increased 3':5'-cyclic GMP (cGMP) levels. Under the assay conditions used (2 x 10(8) cells per ml in 10 mM phosphate buffer, pH 6.0) cAMP (5 x 10(-8) M final concentration) increased cGMP levels from 1 pmol per 10(7) cells to 7 pmol per 10(7) cells in 10 sec and basal levels were recovered in 20-25 sec. cGMP accumulation did not occur when cells were in the cAMP-insensitive stage. cAMP-sensitive cells responded with increased cGMP levels when triggered by 5 x 10(-8) M 5'-CH(2)-cAMP or 10(-5) M adenosine-5'-methylmonophosphate (5'-AMPMe) but not after addition of 5 x 10(-8) M 3':5'-cyclic IMP (cIMP) or 5 x 10(-8) M 5'-AMP. As agonists of cAMP, 5'-CH(2)-cAMP and 5'-AMPMe have, respectively, more than 10% and 1% the chemotactic activity of cAMP, while cIMP has 0.01% the activity of cAMP and 5'-AMP is inactive up to a concentration of 10(-3) M. cAMP-mediated cGMP formation was dependent upon cAMP concentration, with a half-maximal cAMP concentration of about 10(-8) M. This cAMP concentration agrees closely with that necessary for half-maximal receptor occupation. cAMP-mediated cGMP formation was independent of the presence of extracellular Ca(2+); cell aggregation and chemotaxis were also independent of the presence of external Ca(2+). Therefore, cAMP action does not depend on stimulation of the Ca(2+) influx. cAMP was found to mediate desensitization of cAMP-dependent cGMP formation. Addition of 5 x 10(-8) M cAMP to sensitive cells induced a desensitization period that lasted 1-5 min. Desensitization was dependent on the cAMP concentration. Finally, we propose that the translation of a chemotactic signal from the cell surface to pseudopod formation in Dictyostelium involves changes in the levels of cGMP.


Effects of thyrotropin on the thyroid cell membrane: hyperpolarization induced by hormone-receptor interaction. Cultured thyroid cells accumulate the lipophilic cation triphenylmethylphosphonium, indicating that there is an electrical potential (interior negative) across the plasma membrane. Thyrotropin stimulates the uptake of the lipophilic cation 3-fold, and the proton conductor carbonylcyanide-m-chlorophenylhydrazone causes efflux of triphenylmethylphosphonium accumulated in the presence or absence of thyrotropin. The stimulatory effect of thyrotropin on triphenylmethylphosphonium accumulation is not mimicked by human chorionic gonadotropin, a glycoprotein hormone with a similar structure whose target organ is not the thyroid, and the effect is abolished if the thyrotropin-receptor activity of the cells is destroyed by treatment with trypsin. Analogous effects are observed with thyroid plasma membrane vesicles which are essentially devoid of mitochondrial and soluble enzyme activities. Triphenylmethylphosphonium uptake and stimulation by thyrotropin occurs when NaCl, KCl, or Tris.HCl concentration gradients are artifically imposed across the vesicle membrane ([salt](out) > [salt](in)). It seems likely, therefore, that triphenylmethylphosphonium uptake is driven by a chloride diffusion potential (interior negative) and that thyrotropin either increases the permeability of the membrane to anions or decreases its permeability to cations. Thyrotropin-stimulated triphenylmethylphosphonium uptake in the vesicle preparations reaches a quasi steady-state within 3 min; in contrast, thyrotropin-stimulated adenylate cyclase activity is negligible during this period of time, becomes measurable after about 4 min, and is optimal after 12-15 min. Thus, a primary mode of action of thyrotropin on the thyroid cell may be an alteration in the electrical potential across the plasma membrane. The relevance of this observation to the mechanism of action of other glycoprotein hormones, certain bacterial toxins, and interferon is discussed.


Degradation of cellular mRNA during infection by herpes simplex virus. The fate of preexisting mRNA sequences was examined after infection by herpes simplex virus. Murine erythroid cells transformed by Friend leukemia virus were used as the host. Such cells, when exposed to 2% dimethyl sulfoxide, produce large amounts of globin and globin mRNA. The protein and its mRNA are easily recognized at 4 days by electrophoresis in high percentage acrylamide gels and by hybridization to cDNA, respectively. Herpes simplex virus replicates in these cells. By 2 hr after infection the rate of protein synthesis decreases to 30% of the level in mock-infected cells and only 49+/-8% (SEM) of the globin mRNA sequences present prior to infection could be detected by hybridization to cDNA. At 4 hr after infection, when the rate of protein synthesis in infected cells is at a maximum, only about 15% of the globin mRNA sequences remained. Control experiments support the hypothesis that globin mRNA sequences are degraded after infection by herpes simplex virus.


Fidelity of synthesis of preribosomal RNA in isolated nucleoli and nucleolar chromatin. Comparisons were made of the T1 ribonuclease digests of 32P-labeled nucleolar 45S RNA of intact Novikoff hepatoma cells and the RNA synthesized in vitro by isolated nucleoli. Approximately 200 oligonucleotide spots were found in the two-dimensional chromatogram of 45S nucleolar RNA labeled in vivo, which includes fragments of 18S and 28S rRNA and nonconserved spacer regions; four spots containing 2'-O-methyl nucleotides were not found in the corresponding pattern of RNA labeled in vitro. This high degree of fidelity was retained in the patterns of spots from the RNA produced with nucleolar chromatin as template. This specific expression of rDNA was lost when the nucleolar chromatin was completely deproteinized. Specific spots found in the control patterns were absent and many nonspecific oligonucleotides were found to be labeled. A similar nonspecific chromatogram pattern was found when nucleolar chromatin was transcribed with RNA polymerase (nucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) of Escherichia coli. These results show that specificity of genetic expression in vitro of isolated chromatin of eukaryotic systems is dependent on the chromatin-associated proteins and the type of RNA polymerase present.


Transformation by simian virus 40 induces virus-specific, related antigens in the surface membrane and nuclear envelope. Nucleus- and mitochondrion-free membranes from hamster lymphocytes transformed by simian virus 40 (SV40), GD248 cells, cause guinea pigs to produce immune sera that reveal the presence in GD248 plasma membranes and mitochondria of two types of glycoprotein that are not detected in membranes of normal lymphocytes [Schmidt-Ullrich, R., Thompson, W. S. & Wallach, D. F. H. (1977) Proc. Natl. Acad. Sci. USA 74, 643-647]. Indirect immune fluorescence of living, SV40-transformed T19 hamster reticulum cells, Balb/c 3T3 mouse fibroblasts, and W18 VA2 human fibroblasts, using the antisera against GD248 membrane, at 4 degrees produced a distinct cell surface fluorescence; however, above 20 degrees , staining at the nuclear perimeter, the SV40 U-antigen reaction, becomes equally prominent. In SV40-transformed cells that had been fixed in cold acetone, as well as in purified GD248 nuclei, thermostable U-antigen staining is dramatic, but there is no reaction for nuclear T-antigen. Rabbit antisera against T19 cells gave immunofluorescence reactions equivalent to those obtained with the antisera against GD248 cells. Normal guinea pig or rabbit sera and cells that had not been transformed by SV40 gave no reaction. Our sera from tumor-bearing hamsters gave only nuclear T-antigen fluorescence. The results indicate the presence of related, SV40-specific antigens in the surface membranes, nuclear envelope, and possibly other intracellular organelles of SV40-transformed cells.


The intestine as a source of apolipoprotein A1. The major apoprotein of rat mesenteric lymph chylomicrons has been isolated and characterized and shown to be identical to apoprotein A1 (apo A1) isolated from serum high density lipoprotein (HDL). During intestinal lipid absorption, active synthesis of apo A1 was demonstrated by radioactive amino acid incorporation into lymph chylomicron A1 as well as lymph HDL. Immunofluorescence studies of intestinal epithelium demonstrated a marked increase in apo A1 fluorescence, confirming an active synthesis of this apoprotein during lipid absorption. Quantitative immunoelectrophoretic methods were used to measure apo A1 in lymph and peripheral blood during various conditions designed to estimate the quantitative importance of intestinal apo A1 to the levels of circulating lipoproteins. During lipid feeding there was an increase in lymph apo A1 that was associated with lymph lipoproteins (50%) of density less than 1.006 g/ml whereas in basal lymph most apo A1 (85%) was in the lipoproteins of density greater than 1.006 g/ml. Lipid feeding in animals without lymph fistulas resulted in a significant increase in serum apo A1 levels; biliary diversion, designed to eliminate intestinal lipoproteins of density less than 1.006 g/ml, resulted in a significant decrease in serum apo A1 levels. These studies demonstrate that the intestine actively synthesizes apo A1 and is a significant source of this apoprotein for circulating lipoproteins.


Enhanced oncogenic behavior of human and mouse cells after cellular hybridization with Burkitt tumor cells. Studies were made of the expression of the Epstein-Barr virus (EBV) in somatic hybrids of Burkitt tumor cells and human or mouse cells to determine whether EBV genetic information associated with the capacity to transform leukocytes of human and non-human primates could be maintained and expressed in nonlymphoblastoid cells. Data obtained thus far suggest that at least one characteristic associated with cellular transformation (loss of contact inhibition) is expressed only in nonlymphoblastoid cells in which the EBV genome is maintained. In addition, we have demonstrated that human epithelial/Burkitt hybrid cells (D98/HR-1 and D98/Raji) are more oncogenic in nude (athymic) mice than are cells of the human epithelial parental line, D98, or one of the Burkitt lymphoblastoid parent cell lines (Raji); the HR-1 Burkitt parent cell line was as oncogenic as the hybrid cell lines but the time required to induce tumors was much longer. Thus, human epithelial cells show alteration of growth properties in vitro and in vivo after cellular hybridization with Burkitt tumor cells.


Cholinergic metabolism and synapse formation by a rat nerve cell line. The PC12 clone of a rat pheochromocytoma is able to synthesize acetylcholine. The amount of acetylcholine synthesized, and the specific activity of choline O-acetyltransferase (acetyl-CoA:choline O-acetyltransferase, EC 2.3.1.6), varies as a function of the culture growth curve and is dependent on cell density. The specific activity of choline acetyltransferase is increased by nerve growth factor, growth-conditioned medium from a variety of cell types, and adenosine 3':5'-monophosphate. Finally, the PC12 cell line is able to form cholinergic synapses with a clonal cell line of skeletal muscle origin.


Effects of electroshock and drugs administered in vivo on protein kinase activity in rat brain. The effect of electroshock and treatment with reserpine, amphetamine or lithium chloride on protein kinase activity in synaptic membrane fragments prepared from rat brain was investigated. Naive rats subjected to electroshock procedures showed significant increases in both basal and cyclic AMP-stimulated activity irrespective of whether the treatment was sham, acute or chronic. These increases did not occur in animals which had been tamed by daily handling for 15 days prior to treatment, suggesting that the response was induced by the stress of an unfamiliar situation. Administration of lithium chloride and reserpine caused a small but significant increase in the stimulated activity. Doses of d-amphetamine of 5 mg/kg had no effect on either basal or stimulated activity, but higher doses (up to 15 mg/kg) resulted in a pronounced increase in both activities, which may have been related to drug-induced stress.


Evidence for nuclear sites of stereospecific opiate binding in neuroblastoma cells in continuous culture. A class of stereospecific, saturable receptors for narcotic drugs has been found in a clone of mouse neuroblastoma cells in continuous culture. Cells grown in the presence of 10(-8) M etorphine, a synthetic opiate, had an increased doubling time. The neuroblastoma cell nucleus was found to be the sole site for the high affinity binding of etorphine. It was found that these nuclear receptors (1) became saturated at a concentration of 2 X 10(-9) M etorphine, (2) had a dissociation constant of 5 X 10(-11) M etorphine, (3) were capable of binding etorphine stereospecifically at 37 degrees C but not at 4 degree C, and t4) were protein in nature as evidenced by treatment with proteolytic enzymes. More importantly, the receptor activity appeared to be chromatin-associated.


Cellular metabolic control by chemical modification of cell membrane. Various reagents used in the chemical modification of amino- and carboxy-groups of proteins, and of carbohydrates of glycoproteins and glycolipids, inhibit respiration in ascites tumor cells concomitant with release of potassium ion from those cells. The respiratory activity of washed ascites tumor cells is increased by exogenous addition of potassium ion. The lowered respiratory control index as well as oxidative phosphorylation of aged mitochondria are restored upon increasing the potassium concentration of the incubation mixture in the presence of respiratory substrates. The data suggest that the potassium ion level of cells is changed by modifying physicochemical properties of membrane components and that cellular energy metabolism is regulated by intracellular potassium ion concentration.


Possible involvement of 5-hydroxytryptamine and cyclic-AMP in tolerance to tremorine analgesia in mice. The phenomenon of tolerance to the analgesic action of tremorine in mice was studied by the hot-plate and tail-clip methods. Reduction in 5-HT levels in brain by parachlorophenylalamine pretreatment decreased the ED50 of tremorine analgesia in tremorine tolerant mice. 5-Hydroxyptophan, L-Dopa or alpha-methyl-para-tyrosine did not influence the analgesic response to tremorine in tremorine tolerant animals. However, theophylline was found to enhance the tolerance to tremorine analgesia. Brain 5-HT and cAMP are probably involved in tremorine tolerance, whereas neither noradrenaline nor dopamine is involved in the phenomenon.


Sites of action of morphine involved in the development of physical dependence in rats. III. Autoradiographic studies. Morphine withdrawal was precipitated by injection of 3H-naloxone into restricted parts of the ventricular system of rats made tolerant to and dependent on morphine by repeated pellet implantation. The spread of the drug was evaluated by autoradiography and compared with the withdrawal signs precipitated in the same experiment. When the antagonist could spread into the tissue surrounding the 4th ventricle and the caudal parts of the aqueduct (penetration depth about 1.5 mm), a strong withdrawal syndrome was displayed. In contrast, only weak or no withdrawal signs were observed when the spread of naloxone was restricted to the surroundings of the lateral ventricles, the 3rd ventricle, and the rostromedial parts of the aqueduct. The same was true when the spread of the antagonist was limited to the ventral surface of the brain stem. It is concluded that structures located in the anterior part of the fossa Rhomboidea, and possibly also in the caudal part of the periaqueductal grey matter, are sites for the development of physical dependence on morphine giving rise to the withdrawal signs studied in these experiments.


Incidence of bone metastases in women with minimal and occult breast carcinoma. A total of 42 women with proved minimal (14) or occult (28) breast carcinoma were evaluated for up to 48 months (mean = 28 months). Bone scans, radiographs, blood chemistries, enzyme levels, and operative findings demonstrated metastases only in those 6 patients (14%) who presented with symptomatic metastases from an unknown primary source. The other 36 women are free of metastases. Preoperative radionuclide bone scans may not be necessary for women with minimal and asymptomatic occult breast cancers.


What causes lower neck uptake in bone scans? Lower neck uptake, frequently seen on anterior views of bone scans done with 99mTc-phosphate compounds, was studied in 122 patients to determine its incidence and etiology. Increased uptake was identified in the lower neck anteriorly in 46 patients (38%). In 14 of these, moderately severe to severe arthritis appeared to cause the uptake; metastatic disease was the cause in 8 cases; and in 1 case there was prominent uptake in the thyroid cartilage. Positioning artifact accounted for the uptake in the other 23 cases. Thyroid uptake was never observed.


Radiography and scintigraphy in tumoral calcinosis. A case of tumoral calcinosis with tumoral deposits about the knee is reported. Radiographs demonstrate juxta-articular calcific deposits about the right second and fifth distal interphalangeal joints and the left first distal tuft, both hips, both shoulders, first distal phalanx of the left foot and right knee. Radionuclide images clearly demonstrate all areas of tumoral calcinosis.


A comparative evaluation of a case of cystosarcoma phylloides: ultrasound, xeroradiography and thermography. The results of xeroradiography, thermography, and ultrasonic gray scale echography in a case of cystosarcoma phylloides are presented. Echography better described the lesion than the other techniques. Gray scale echography displays the infrastructure of the breast tissues and differentiates between types of soft tissue.


Chromatographic patterns of prostaglandins using different lots of silicic acid. The chromatographic patterns of prostaglandin E2 chromatographed on various lots of silicic acid are presented. Two lots of Mallinckrodt silicic acid tested give totally unsatisfactory separation of PGE2. Sigma silicic acid SIL-R gives better separation than any of the Mallinckrodt lots tested. Increasing the size of the "E" fraction of solvent can give satisfactory separation on Mallinckrodt silicic acid.


Inhibition of prostaglandin synthesis in mouse 3T3 fibroblasts and human platelets by substituted phenols. Several substituted phenols with antioxidant properties were potent reversible inhibitors of prostaglandin synthesis in 3T3 cell cultures. The ID50's for prostaglandin (PG) E2 synthesis in these cells were 0.1 muM for 2,6-xylenol, 5 muM for tricresol, 6 muM for p-cresol, 7 muM for o-cresol, 15 muM for 3,5-xylenol, 30 muM for m-cresol and 100 muM for phenol. The corresponding values for aspirin and indomethacin were 4 muM and 0.02 muM, respectively. The substituted phenols also inhibited serotinin release, aggregation and prostaglandin synthesis in human platelets induced by arachidonic acid but not by PGG2.


[Malignant hepatoma (clinical, biological and morphological comparisons)]. The authors make an analysis of 8 cases of malignant hepatoma discovered in 20 362 surgical patients between 1965 and 1975. The analysis of these cases allowed to conclude as follows: 1). In the presence of chronic digestive disturbances, or of hepato-splenomegaly, accompanied by progressive deterioration of the general condition of the patient, and of the use of large amounts of analgetic drugs, one should consider more than usually the possibility of hepatic neoplasia. 2). The first element to be considered in the frame of diagnosis of malignant hepatomas should be the morphological one. Of practical value in the diagnostic are the following: hepatic scintigraphy, angiography and bioptic laparotomy. 3). Of the conventional biologic tests (including the immunological tests), evaluation of serum alpha-foetoprotein is the only one with an orientative diagnostic value.


Cyclic Amp and the release of antral gastrin. The isolated, vagally innervated antral pouches in anesthetized dogs were irrigated with 095% acetylcholine chloride (Ach) or a 0.4 M mixture of non-essential amino acids (AA), or distended at 30 cm H2O pressure. In addition,2-deoxy-D-glucose (2-DG) was administered intravenously. Significant increases in antral venous gastrin, caused by gastrin releasers, were associated with a significant rise in cAMP content in the antral mucosa. Acidification of the antrum with simultaneous administration of 2-DG, and acidified AA abolished gastrin release, and significantly reduced mucosal cAMP. Acidified Ach and distension of the antrum with 0.1 N HC1 supressed gastrin levels without significantly changing the mucosal cAMP content. Since the augmented gastrin release was accompanied by an increased mucosal cAMP, the results suggest that cAMP participates in the action of gastrin secretagogues.


Mechanisms of the H-2 effect on viral leukemogenesis. Evidence has been gathered which supports the notion that two distinct but interacting mechanisms, controlled by loci mapping within the H-2 complex, influence Friend murine leukemia virus (FV) disease. One mechanism, controlled by a gene mapping in or close to H-2D, influences the capacity of the H-2D gene product to form molecular complexes with FV molecules in the plasma membrane of infected cells. Formation of a complex appears to provide a target antigen for syngeneic cytotoxic T-lymphocytes, to cause co-capping of FV and H-2D antigens, to permit the selective inclusion of H-2Db molecules into progeny Friend virions, to influence the long-term maintenance of virus production in vitro and, in conjunction with the second mechanism, to stimulate the generation of cytotoxic T-lymphocytes. This second mechanism is controlled by a gene in the H-2K or H-2I region, and, in the presence of an H-2/FV molecular complex immunogen, influences the generation of H-2 restricted cytotxic T-lymphocytes and the rate of rejection of syngeneic FV-induced tumor cells.


[Familial cancer syndrome studies in 4 generations of a family]. An account is given of a family from the Canton of Valais suffering from hereditary adenocarcinomatosis. The pedigree extends over four generations; the first three comprised 47 individuals (28 males, 19 females), of whom 21 (16 males and 5 females), i.e. 44.6%, have malignant tumors. Of the 32 people in the fourth generation, only one individual is affected to date (a girl age 21, IV/4). There were 27 tumors in all: 16 adenocarcinomas of the colon, two gastric adenocarcinomas, one duodenal adenocarcinoma, one rectal adenocarcinoma, one papillary carcinoma of the ovary, one osseous sarcoma, one cutaneous fibrosarcoma, a multiform glioblastoma of the basal nuclei of the brain, a basocellular epithelioma, a cerebral metastasis from an adenocarcinoma, the origine of which has not been established, and a tumor invading the biliary tract. Three members of the family had multiple tumors. In three of the patient the colonic adenocarcinoma was accompanied by one or two polyps. The average age at onset for all tumors was 45 years. It was definitely lower in the third than the second generation (anticipation). The transmission was autosomal dominant, with predilection for the male sex (57.1% male and 26.3% female patients). The penetrance was about 80%. Finally, the diagnostic criteria for hereditary adenocarcinoma are discussed and the different familial forms of cancer are reviewed.


Correlation between lipid synthesis in tumor cells and their sensitivity to humoral immune attack. Prolonged incubation of two antigenically distinct, chemically induced guinea pig hepatomas with relatively high concentrations of chemotherapeutic drugs or metabolic inhibitors increases their susceptibility to killing by antibody and complement. This effect is reversible when the cells are cultured in the absence of the drugs. The drug-induced sensitivity and the ability of the cells to recover their resistance to killing are directly correlated to their ability to synthesize complex lipids.


Dopamine and adenosine 3',5'-monophosphate responses of single mammalian sympathetic neurons. Acetylcholine (ACh), dopamine, and dibutyryl-adenosine 3',5'-monophosphate (dbcAMP) were applied iontophoretically to the rabbit superior cervical ganglion cells from triple-barreled micropipettes, and the response was recorded by intracellular techniques. All ganglion cells tested responded to the depolarizing action of ACh, whereas less than half of the cells that responded to ACh were hyperpolarized by dopamine. This effect was blocked by low concentrations of haloperidol. None of the cells examined responded to dbcAMP applied by iontophoresis. Hence, the present result is not consistent with the concept that a cyclic AMP mechanism underlies the hyperpolarizing effect of dopamine.


Conformations of prostaglandin F 2alpha and recognition of prostaglandins by their receptors. The conformation of prostaglandin F 2alpha (PGF 2alpha) has been determined by x-ray diffraction techniques. Two independent conformers of PGF 2alpha, studied as the tris(hydroxymethyl)methylamine salt, are observed to adopt the familiar "hairpin" conformation with the alpha and omega chains aligned roughly parallel. The conformers differ in ring conformation and at the C(17)-C(18) bond, one adopting a C(9) envelope ring conformation and a trans geometry at the C(17)-C(18) bond, while the other adopts a C(8) envelope ring conformation and a novel gauche geometry about C(17)-C(18). Comparison of the conformation of PGF 2alpha with that of prostaglandin E2 suggests a recognition mechanism which would permit PGF 2alpha and prostaglandin E receptors to distinguish between the two potent prostaglandins. The recognition model explains much of the binding data for the PGF 2alpha receptor in the corpus luteum and predicts the existence of an interesting PGF 2alpha analog.


Thyroid hormone action: the mitochondrial pathway. The subcellular compartments have been investigated to compare proteins capable of binding triiodothyronine and thyroxine; specific binders have been found in cytosol, nuclei, and mitochondria from rat liver and kidney. The binding protein from the inner mitochondrial membrane had the highest association constant (greater than 10(11) liters per mole), suggesting possible direct hormone action on the mitochondria. Binding of hormone analogs was found to be related to known physiological potency, and stereospecific discrimination between L- and D-thyroxine was observed. The saturable receptor was found in the mitochondrial membranes of rat liver, kidney, myocardium, and skeletal muscle but not in mitochondria from the unresponsive tissues: brain, spleen, and testis. Oxidative phosphorylation by mitochondrial vesicles from hypothyroid rats increased after the addition of physiological concentrations of triiodothyronine, which corroborated direct hormone action on mitochondria.


Axial bending in the early chick embryo by a cyclic adenosine monophosphate source. A microelectrode continuously releasing cyclic adenosine monophosphate can divert the axis of the early chick embryo and attract cells on its ventral surface. Cell movement in the intact embryo may be controlled by a cyclic adenosine monophosphate signal.


Rous sarcomas in chickens: enhanced growth coexisting with concomitant immunity. Chickens bearing Rous sarcoma virus-induced tumors in one wing did not develop new tumors when subsequently inoculated with Rous sarcoma virus in the other wing. However, the second inoculation of Rous sarcoma virus caused accelerated growth of the established tumors. This phenomenon was found to be bursa-dependent. Paradoxically, established tumors in bursectomized chickens grew at a diminished rate if the chickens were reinoculated with Rous sarcoma virus.


[Acute oligoanuric renal insufficiency in malignant nephroangiosclerosis]. The concept of acute renal failure with anuria due to malignant nephro-angiosclerosis, is uncommon. We report two cases and compare them with others in the world literature. Knowledge of this disease entity is of triple interest: In diagnosis and classification in the field of acute vascular nephropathy with anuria. We cannot emphasise too much the interest of early renal biopsy after correction of abnormal blood pressure. In physiopathology, these malignant nephro-angioscleroses give rise to hypertension of pressor type with high renin levels. Finally therapeutic, for there exist drugs adapted to this type of hypertension. The association of acute renal failure and malignant nephro-angiosclerosis should be treated as an emergency, to avoid the passage to terminal and irreversible renal failure.


[Radiologic study of arterial calcifications of the lower extremities in diabetic patients]. The authors analyse the frequency of arterial calcifications in the lower limbs in diabetics. These calcifications were found in the thighs and legs and also in the feet. They were more frequent in diabetics than in normal subjects and occurred earlier in life. Their presence showed the existence of a severe microangiopathic process.


[Initial trial of a new test for the evaluation of hepatocellular function: the TMB test]. The oral administration of 400 mg of trimethoxy 1-3-5-benzene or T.M.B. is normally followed by a high urinary excretion of dimethoxy, 1, 3, hydroxy 5 benzene and other metabolites mainly in the form of conjugates. Urinary excretion during the first 3 hours is strongly reduced in liver disease. By a simple blind test we showed that the test is positive in subclinical and laboratory-negative liver failure.


[Intra-articular iron sequestration and rheumatic anemia. Histologic and isotopic study]. The authors explored 48 patients with various forms of inflammatory rheumatism and 8 controls. The exploration consisted of seeking in the synovial membrane ferric pigments by Perls stain, or studying the movements of Fe 59 bound to siderophyllin in the joints. Intra-articular iron deposits were thus demonstrated only in the inflammatory cases. Its mechanism is discussed. It seems sufficient to be the cause of rheumatic anemia which is thus different from other forms of inflammatory anemia owing to the importance of the iron uptake.


[Immunologic status of women of western Brittany with regard to the Torch complex (toxoplasmosis, rubella, cytomegalovirus, herpes); necessity of a polyvalent serologic surveillance in pregnancy]. We have carried out a sero-epidemiological investigation with regard to the "Torch complex" in 702 women aged from 11 to 49 years and living in an urban district of the Finistere or Morbihan. The proportion of protected women was relatively low compared with other regions of France: 50.14% for toxoplasmosis 74.64% for rubella, 23.70% for cytomegalovirus and 39.61% for herpes. Furthermore, no woman had any antibodies for the virus of lymphocytic choriomeningitis, another potentially teratogenic virus. It thus seems advisable to supervise the serology of pregnant women in our region for the whole of the Torch complex. The practical modalities and the choice of serological reactions are discussed.


Acute haemorrhagic conjunctivitis in Hong Kong 1971-1975. Acute haemorrhagic conjunctivitis occurred in epidemic in Hong Kong in 1971 and 1975 respectively. Two picornaviruses, the Enterovirus type 70 and an antigenic variant of Coxsackie virus type A24, were isolated during the 1971 outbreak, while only the latter virus was detected in the 1975 outbreak. In 1972 and 1974, sporadic cases of conjunctivitis associated with Enterovirus type 70 were observed. Serological survey in late 1975 showed a high proportion of the population might still be susceptible to both viruses. Recrudescence of the disease is conceivable when environmental condition is favourable for the spread of these viruses.


Alpha-chain disease. A case report with autopsy findings. A 19-year-old Coloured man with alpha-chain disease died after perforation of a lymphoma in the duodenojejunal region. At autopsy he was found to have a dense plasma cell infiltration of the lamina propria of the entire small bowel, causing thickening of that organ, and a malignant lymphoma in the duodenojejunal region which had spread to regional mesenteric nodes and to the liver only.


The effect of practolol and butoxamine on aortic arch malformation in beta adrenoreceptor stimulated chick embryos. An equimolar dose of the beta-1 adrenoreceptor antagonist practolol administered to embryonic chicks prevents the induction of aortic arch malformations by isoproterenol. Whereas 3.75 X 10(-9) mole isoproterenol in 5  microliter saline solution induced aortic arch anomalies in 39% of embryos injected at Hamburger-Hamilton developmental stage 26, pretreatment with practolol one to two minutes before catecholamine administration reduced the anomaly rate to to 4%. Practolol when injected alone did not influence survival rate nor did it cause cardiovascular malformations. Probably the most significant result of this study involves the prevention by practolol of aortic hypoplasia and interrupted aortic arch complexes, anomalies frequently induced by isoproterenol when administered at this stage of embryonic chick development. Butoxamine, a beta-2 adrenoreceptor antagonist, did not block the overall effect of isoproterenol nearly as effectively as did practolol. Results from the present study suggest that aortic arch anomalies may be induced in embryonic chicks via beta-1 adrenoreceptor stimulation. Beta-2 receptor stimulation does not appear to be as significantly involved.


ADPase activity of normal and atherosclerotic human aorta intima. ADP plays a key role in platelet aggregation and the enzymatic removal of this nucleotide may be important in the pathogenesis of intravascular thrombosis and atherosclerosis. Aortic intima extracts have ADPase activity and is able to remove small quantities of ADP efficiently. ADPase activity was assayed by measuring the catabolism of 2 micrometer 14C-ADP (final concentration) by the tissue extracts. Extracts prepared from normal, moderately and severely atherosclerotic human aortic initimas showed a significant progressive decrease in ADPase activity with increasing atherosclerosis. ADPase activity of the arch, thoracic and abdominal regions of normal aortas did not vary significantly, and thus did not correlate with the anatomical distribution of atherosclerosis. Vascular ADPase activity seems relevant in thrombogenesis since it may be a link between blood platelets and blood vessel wall interaction.


Transmissible gastro-enteritis of swine in the netherlands the application of the direct fluorescent antibody techique for diagnosis. The epizootiology of transmissible gastro-enteritis (TGE) of swine in 1975 and 1976 in the Netherlands was studied. Diagnosis was made by the direct fluorescent antibody technique (FAT). Specimens from the small intestine of suspected animals were examined for this purpose. The results obtained by the direct FAT were proved to be reliable, if the specimens were derived from acutely sick animals and examination could be carried out as soon as possible. The results confirmed that TGE occured in any season of the year. The peaks of epizootic outbreaks were in the spring and winter of those years.


Ultrastructural study of immunoblastic lymphadenopathy. An autopsy case of angio-immunoblastic lymphadenopathy with dysproteinemia (Frizzera et al. 1917) or immunoblastic lymphadenopathy (Lukes and Tindle 1975) is reported. Clinical pictures and morphologic characteristics of affected organs were typical of this disease. In spite of combination chemotherapy, the patient took a rapid fatal course. Post-mortem examinations disclosed involvement of the lymph nodes, liver, lungs, kidneys and skin. Cellular infiltrates in the kidney were more monomorphous, suggesting the potential for the development of immunoblastic sarcoma. Electron microscopies of the affected lymph nodes revealed the proliferation of immunoblasts characterized by moderate amount of clear cytoplasm with abundant polyribosomes and by large nuclei with prominent nucleoli. Undulated tubules associated with the endoplasmic reticulum and giant mitochondria with the centrally placed cristae were observed in occasional immunoblasts. Cytoplasmic fragments of immunoblasts and filamentous material among the cells were considered to correspond to the amorphous intercellular material seen in histologic sections.


Neurocristopathies in African subjects.

Neurocristopathies in African subjects. The distribution of neural crest cells is briefly reviewed. The behaviour of neural crest derivatives in Africans is different from that seen in Caucasians, both in terms of physiology and pathology. The most obvious variation is in the depth of pigmentation of the skin, but most diseases of neural crest derived tissues show differences between Black and White subjects. The reason for these differences are not readily apparent but appear to be of genetic rather than environmental origin.


[Studies on the Simulium damnosum complex in West Africa - V. Morphological identification of the females in Ivory Coast (author's transl)]. The morphological identification of adult females of the Simulium damnosum complex is essential for studies on the ecology and vector potential of the species concerned. Detailed morphological studies of adult females obtained at breeding sites, where larvae had been identified cytotaxonomically, allowed the establishment of an identification key. This key, based on morphological characters of the antennae and maxillae, is dealing with the six species of the complex known to occur in the Ivory Coast. It has been used, with success, during ecological and epidemiological studies in the field.


[Characteristics of the upper cell surface in cultures of normal and SV-40 virus-transformed epithelium of mouse kidney. Study with the aid of scanning electron microscopy]. Central cells of the normal epithelial sheet are sparsely covered by microvilli. Numerous microvilli were seen in the regions of intercellular contacts. Marginal cells of sheets had a finely developed lamellar cytoplasm (lameloplasm) with smooth upper surface at their free margins. A transformed cell line (MPTR) resembled normal parent cells by its ability to form monolayered sheets in cultures. More microvilli of increased length appeared on the upper surface of central MPTR cells. The normal structure of lamelloplasm was changed at the free edge of the MPTR sheets. It is suggested that abnormal cell attachment to the substratum may be responsible for the altered cell surface morphology (increased length of microvilli, defective, structure of lamelloplasm) in the MPTR cultures.


[Study of structural changes of contractile muscle proteins with the aid of polarization ultraviolet fluorescence microscopy. 1. Conformational changes of F-actin in the muscle fiber caused by ATP and its analogs]. Increase of anisotropy of F-actin fluorescence of balanus and rabbit muscle fibers under the influence of ATP, AMP and pyrophosphate in EGTA presence was detected by means of the polarized ultraviolet (UV) fluorescent microscopy methods. The fluorescence anisotropy changes are assumed to be associated with the conformational changes in the actin. ATP cause more noticeable changes of actin structure, than pyrophosphate and AMP. The conformational changes in the actin of balanus and rabbit muscle fibres were similar. ATP and its analogs induced also decrease of UV fluorescence anisotropy of A-band which appears to be associated with conformational changes in myosin. It was siggested that the changes in fluorescence of anisotropy of A-bands are due to structural changes in both HMM and LMM parts of myosin molecule.


[Analysis of substances issuing from Zajdela ascitic hepatoma cells after exposure to UV radiation of different wave-lengths. I. Relationship to dose]. The release of substances from the Zaidela ascitic hepatoma cells after irradiation with physiological doses of short-wave (254 nm) and long-wave (300-380 nm) UV light (far and near UV light) has been studied spectrophotometrically. Within the range of 200-520 nm, the absorption spectra of releasing substances show maxima at 215 and 260 nm and are identical to spectra of non-irradiated cells. The amount of substances increases with dose making up, at the maximal alteration, 180-220%, of the amount releasing from non-irradiated cells. Irradiation with far UV light exceeds by one order that with near UV light. The effect of minimum doses is opposite to the action of high doses: the release of substances from irradiated cells is much less.


[Karyological study of the HTR-5 cell line]. A karyologic study of the human fibroblasts transformed by the SV-40 virus at the level of the 15th subculture shows that this line (HTR-5) consists mainly of cells with a pseudodiploid karyotype. The cells of the investigated line contain marker chromosomes characteristic for this cell culture.


[Participation of some nucleotides in regulation of phosphoprotein phosphates activity in rat and chicken myocardium]. Adenine- and uridine di- and triphosphates (in a 3 mM concentration) increase considerably phosphoprotein phosphatase (PPPase) (EC 3.1.3.16) activity of rat and chicken myocardium homogenates. AMP and Pi are effective inhibitors of the enzyme. The ATP activating effect is also shown in partially purified preparations of rat myocardium PPPase. ATP is able of protecting significantly the enzyme during its thermodenaturation.


[Effect of insulin on activity of carbohydrate metabolism enzymes in loach embryos in early development]. It is established that in embryos incubated until the early blastula stage in the solution of insulin with addition of cycloheximide or puromycin, there is neither a decrease in the hexokinase and glucose-61 phosphate dehydrogenase activities nor an increase in the phosphofructokinase activity, as it is shown under the influence of insulin only. Puromycin removes an inhibitory effect of insulin on the glucose-6-phosphatase activity, and actinomycin D removes this influence with respect to glucose-6-phosphate dehydrogenase and glucose-6-phosphatase activities. The addition of antibiotics removes inhibition of the hexokinase, glucose-6-phosphate dehydrogenase and glucose-6-phosphatase activities by the hormone in the unfertilized eggs as well. Actinomycin D alone inhibits the hexokinase and activates the phosphofructokinase activities in the embryos and eggs, puromycin decreases their hexokinase activity and cycloheximide has the same effect on the glucose-6-phosphatase activity in the embryos only.


[Characteristics of protein kinases from myocardium and their use for studying cAMP content in rat tissues after muscular activity]. Protein kinase from the bull myocardium tissue was separated by means of the stepped gradient of buffer concentrations on DEAE-cellulose. Sensitivity of the obtained fractions to cAMP was studied. Protein kinase isolated at elution by 0.3 M potassium-phosphate buffer from DEAE-cellulose is less sensitive to cAMP than protein kinase isolated according to Kuo. A decrease in the content of cAMP is established in the tissues of the skeletal muscles and adrenals of rats after long physical loading. No statistically significant changes are found in the level of cAMP under the same conditions in the myocardium and brain tissues.


Tissue-cultured head and neck tumors: their use in in vitro assays of immune response. Samples of 35 tumors from the head and neck region (25 squamous cell, 2 basal cell, 5 parotid, 3 melanoma, and 1 lymphosarcoma) were cultured after dispersement with either trypsin or collagenase treatment. Growth was established in 14 (40%). Cultured tumor cells were then used as target cells in in vitro assays of patients' cellular and humoral immunity to their own or similar tumors. Preliminary data suggest this may be a reliable method of monitoring responses in patients receiving immunotherapy for head and neck malignancies.


Ovarian tumors and related lesions in aged chimpanzees. The ovaries of a 48-year-old chimpanzee each contained a large bilateral fibrothecoma. A 39-year-old chimpanzee had two small fibrothecomas in one ovary and a well-differentiated Sertoli-Leydig cell tumor in the other; there also was adenomatous hyperplasia of the endometrium. Both animals had extensive thecal hypertrophy in the ovaries. Thecal hypertrophy might have been a source of excessive estrogen and could have been a partial cause of the ovarian tumors.


Encephalomyocarditis virus antibodies in sera from apparently normal pigs. Antibodies which neutralise and precipitate encephalomyocarditis virus habe been found in the serum of more than 28 per cent of normal pigs in Britain. Neutralising activity was found in association with several size classes of antibody.


Experiements with an inactivated hepatitis leptospirosis vaccine in vaccination programmes for dogs. A fluid adjuvanted vaccine consisting of inactivated hepatitis virus (iH) and leptospirae antigens (L) was developed. The vaccine (Kavak iHL; Duphar) was tested in several vaccination programmes both alone and in combination with freeze dried measles (M) or distemper (D) vaccines. The results demonstrate that this new vaccine is also effective in pups with maternally derived antibodies, although a second vaccination at 14 weeks of age is recommended to boost the first vaccination. For the booster vaccination either the iHL-vaccine or the liver attenuated hepatitis vaccine (H) can be used.


Benign human mammary myoepithelioma.

Benign human mammary myoepithelioma. A leiomyoma-like, multifocal tumour developed from intraductal papillomatosis in the breast of a 42 year old woman. The spindle-shaped tumour cells were examined by light, electron and polarizing microscopy, which revealed that they originated from immature "precursor" myoepithelial cells. The author suggests that the tumour be called "myoepithelioma". From the morphological characteristics of the tumour the myoepithelial cells appear to be capable of producing leiomyoma-like benign or malignant tumours. The role that has been attributed by some to the myoepithelial cell in the production of epithelial tumours is problematical, in the light of the present finding.


Electron microscopic study of signet-ring cells in diffuse carcinoma of the human stomach. The signet-ring cells seen in gastric carcinoma of the diffuse type were studied by light and electron microscopy. A classification of these cells into Types A, B and C was developed, based on intracellular mucous granules, nuclei and organelles, and be reactions to PAS and Alcian-blue staining. The gradual transition of type A cells to Type B and the subsequent change of Type B to Type C suggest successive stages in maturation. These cells, especially those of Type B, resemble the mucous neck cells of the corpus and the glands of the pylorus.


[Glyconeogenesis in rat liver in thermal burns]. Processes of glyconeogenesis were increased in rats, subjected to burns. This was corroborated by activation of the glyconeogenesis key enzymes (phosphoenol pyruvate carboxylase and fructose diphosphatase) as well as by the increased incorporation of 14C-amino acids into liver glycogen. The activation of glyconeogenesis had an adaptational importance under burns; it was directed towards the compensation of glycogen content in liver tissue and maintaining of sugar level in blood.


[Nature of the proteinase activated by the contact factor in human blood plasma]. The nature of protamine splitting proteinase, which was formed after treatment of human blood plasma by kaolin or silicone (aerosile), was studied. The activated enzyme did not exhibite the properties of thrombin and plasmin in reactions with specific substrates. Kyninogenic, TAME (N-tosyl-d-,l-arginine methyl ester)-esterase and protamine splitting activities were inherent in the proteinase; these properties enabled to group the enzyme with blood kallikreins. The purified preparations of human blood plasma kallikrein hydrolyzed protamine sulphate even at 10 mcg/ml concentration. alpha2-macroglobulin inhibited the protamine splitting activity of the plasma kallikrein.


[Biochemical heterogeneity of mitochondria from mucosa of rat small intestine]. Mitochondria from mucosa of rat small intestine were separated (0-11% gradient of ficoll) into fractions, differing by the content of protein, DNA and cytochrome a+a3 as well as by the respiratory activity and the activity of succinate- and NADH-dehydrogenases. Content of DNA and cytochrome a+c was shown to be increased in "heavy" mitochondrial fractions. Mitochondria od "midle" fractions possessed the highest functional activity. The data obtained suggest that the mitochondrial preparations are not contaminated by other cytoplasmic structures; they were also not impaired during isolation and fractionation. The activity of RNA and DNA synthesis in vivo and distribution of labelled mitochondrial RNA and DNA in the fractions were studied. The possible relation of mitochondrial heterogeneity to, their biogenesis is discussed.


[Use of circadian rhythms for analysis of the interrelationships between the concentration of glucocorticoids in the blood and the activity of key enzymes for gluconeogenesis in the liver and kidney of rats]. Circadian rhythms of alterations in content of 11-hydroxycorticosteroids and glucose in blood as well as in the activity of phosphoenolpyruvate carboxykinase, fructose-1,6-diphosphatase and glucose-6-phosphatase in liver and kidney tissues were studied under normal and inverted luminous regimens in summer and winter seasons. These patterns were distinctly altered depending on circadian rhythms in the above-mentioned conditions. The maximal content of 11-hydroxycorticosteroids in blood did not correlate with the highest activity of the gluconeogenesis key enzymes in the periods studied. Daily alterations in the activity of phosphoemolpyruvate carboxykinase were shown to be similar to that of the fructose-1,6-diphosphatase in various tissues at different seasons. But glucose-6-phosphatase differed distinctly from these enzymes by the daily rhythm of activity. This suggests that glucose-6-phosphatase has the other mechanism for regulation of its daily rhythm.


[Stimulating effect of glucagon and theophylline on the activity of rat liver betamine-homocysteine-methyltransferase. Role of cyclic adenosine-3',5'-monophosphate]. Effect of glucagon, theophylline, NaF and cyclic 3',5'-AMP on the activity of betaine-homocysteine-methyl transferase (BHMT) was studied in homogenates of rat liver tissue. Intraperitoneal administration of theophylline into adult rats increased the BHMT activity in liver tissue. After administration of theophylline the BHMT activity was distinctly increased in liver tissue within 2 hrs, approaching the maximal value within 3 hrs after which it decreased quickly up to 5-6 hrs. Intraperitoneal administration of glucagon into adult rats also increased the BHMT activity; theophylline, administered simultaneously with glucagon, potentiated the effect of the latter on the BHMT activity in liver tissue. Administration of glucagon into rat embryos 1-2 days before the birth was accompanied by a 2-fold increase in the BHMT activity. In the in vitro experiments theophylline (10(-6)-10(-5) M) showed the stimulating effect on the liver tissue BHMT activity. Dibutyryl adenosine-3,5'-cyclic phosphate and NaF caused the variable effect on the BHMT activity in liver tissue of adult rats. Administration of cyclic-3',5'-AMP (5 mg per 100 g of body weight) decreased the BHMT activity in liver tissue mitochondria and increased 2.5-fold the enzyme activity in cytosole.


[Content of the components of the adenylic acid system in solid and ascitic tumors in the liver of experimental animals]. The authors report the results of separate determination of the concentration of free adenine nucleotides (ATP, ADP, AMP) in tumors, intact animals liver, and tumor-bearing animals liver. In Zajdela ascites hepatoma, ascites tumor NKly and solid lymphosarcoma, solid hepatomas 46 and 22 A the amount of ATP and ADP was found to be markedly reduced compared with their content in the liver. The ratio ATP/ADP is increased in ascites cells of tumor NKly, Zaidela hepatoma and lymphosarcoma and is decreased in solid hepatoma 46 and 22 A. Cell energy potential, calculated on the basis of ATP ratio to a sum of adenine-nucleotides, is also increased in ascites cells of tumor NKly, Zaidela hepatoma and is diminished or remains unchanged in hepatoma 46 or 22A. Cell energy charge is increased in tumor NKly, Zajdela hepatoma, lymphosarcoma and is decreased in solid hepatoma 46 and 22A.


Intensive plasmapheresis as a risk factor for arteriosclerotic cardiovascular disease? Recent studies suggest that the analogy between intensive plasmapheresis and the nephrotic syndrome with respect to plasma protein loss includes events which are generally recognized as risk factors for the development of arteriosclerotic cardiovascular disease. Experimental nephrosis and plasmapheresis alike increase the synthesis of very low density and low density lipoproteins in response to the hypoproteinemic stimulation of plasma protein synthesis. During electrophoresis, these lipoproteins move with the alpha2- and beta-globulins, and the serum levels of these fractions are often elevated in humans subjected to intensive plasmapheresis. Hyperlipoproteinemias of the alpha2- and beta-types are recognized risk factors in the genesis of arteriosclerosis. Another such factor appears to be an increased synthesis of fibrinogen. This protein enhances plasma viscosity and red cell aggregation and may play a role in myocardial infarction. Intensive plasmapheresis thus not only creates an unphysiological and partly depletional state; it is also, like the nephrotic syndrome, capable of producing surplus abnormalities of plasma constituents with a potential for delayed, insidious injury.


[1 of the possible causes of an increase in acid hydrolase activity in homogenates of heart muscle following myocardial infarct]. A "free" activity of acidic hydrolases (acidic phosphatase, acidic ribonuclease and cathepsin D) was increased in homogenates of dog heart muscle with simultaneous decrease of the enzymes activity in the fraction enriched by lysosomes, within 4-5 hrs after ligation of the descending ramus of sinister mitral artery. The adenylate cyclase activity and content of c-AMP were decreased as compared with unaffected part of myocardium. The data obtained suggest that the decrease of the c-AMP content in the impaired region caused a labilization of lysosomal membranes and the secretion of acidic hydrolases into cell cytoplasm.


Electron microscopic studies on the formation of vesicular bodies during cell wall degradation and regeneration in yeast. Vesicular bodies are observed during enzymatic degradation of the yeast cell wall and in the course of wall regeneration. In both cases various types of vesicles are formed and exocytated by different mechanisms. They are characterized cytochemically and their possible role in cell wall regeneration is discussed.


[Controlled clinical study of the herpes antigens LUPIDON H and LUPIDON G]. Results gained in a placebo-controlled, clinical double-blind study with the Herpes antigens LUPIDON H and Lupidon G are reported. Due to treatment with LUPIDON H or LUPIDON G a cure or a more soothed course of the disease and among the indicated effects a prolongation in length of the intervals during which no relapses occurred could be observed with regard to about 80% of all the patients concerned. In opposition to the mentioned effectiveness the treatment success becoming evident after the administration of Placebo under equal test conditions was only evaluated with a rate complying to approximately 30%. The difference determined at those investigations with reference to the efficacy of LUPIDON (80%) in comparison with Placebo (30%) is statistically significant. This outcome confirms the results already observed hitherto in open clinical investigations by other examiners whose findings, too, ascribed to LUPIDON a favourable influence on Herpes simplex diseases in about 80% of treatment cases.


Effect of different amino acids on the phosphatase system of an ectoparasite: Poecilobdella granulosa. Inhibitory effect of different amino acids (L-phenylalanine, L-tryptophan, L-tyrosine, L-histidine-monochloride and L-arginine) on the phosphatase system of the brain, ventral nerve cord, stomach and intestine of the Indian medicinal leech Poecilobdella granulosa, was observed to be substrate, tissue, and inhibitor-specific. Most fascinating observation recorded was the activation of alkaline phosphatase of stomach and intestine by certain amino acids at low concentrations. This has been correlated with the sanguivorous habit of leeches.


Comparative histochemical observations on the excretory system of helminth parasites. The excretory canals of Ascaridia galli (Nematoda) and the protonephridial ducts of Cotylophoron cotylophorum (Trematoda) and Raillietina cesticillus (Cestoda) have been studied with regard to the histochemical localization of lipids, carbohydrates and hydrolytic enzymes. Distinct excretory organs are absent in the acanthocephalan Centrorhynchus corvi. Triglycerides, phospholipids and lipoproteins are seen in association with the wall of excretory canals of A. galli and R. cesticillus, and phospholipids and lipoproteins at the corresponding site in C. cotylophorum. The physiological significance of lipids in association with excretion of substances has been discussed. Low molecular weight glycogen is present in the lumen of excretory canal of A. galli but not in other worms. The common feature of the excretory canals is the presence of enzyme activities of nonspecific alkaline phosphatase and Mg2+-dependent ATPase. Activity of acid phosphatase is seen only in the excretory canals of A. galli. Glucose-6-phosphatase is present in A. galli and C. cotylophorum and absent in R. cesticillus. Weak reaction of 5'-nucleotidase is present in the excretory canals of helminth species studied here. The role of these enzymes in transportation of substances across the wall of excretory canals and also in ionic regulation has been discussed in detail.


Effect of decerebration on the enzymatic system of stomach and intestine of Indian cattle leech, Poecilobdella granulosa. Experiments on decerebration have brought to light an interesting inverse relationship between the phosphatases of two digestive components, at least up to a week. With the activation of all the gastric phosphatases, intestinal phosphatases are inhibited. This has been correlated with metabolic metameric control in leech.


[An additional nerv-lesion in cases of total hip replacement (author's transl)]. Report of early secundary paresis of the sciatic nerve after the implantation of a total artificial hip-joint by the dorso-lateral exposure. The paresis is caused by a cicatrical stenosis of the nerve following a peri- and intranerval bleeding.


[On the enzymatic breakdown of tripolyphosphate and diphosphate in comminuted meat. X. Influence of substrate concentration (author's transl)]. The tripolyphosphatase activity of comminuted bovine muscle increased with rising substrate concentration (up to 0.8% tripolyphosphate in presence of 2% NaCl). Addition of NaCl increased the maximum velocity (Vmax) of the enzymatic hydrolysis, determined by Lineweaver-Burk plots. From the results it is concluded that the product-dissociation is the rate-limiting step in the breakdown of tripolyphosphate. Increasing diphosphate concentrations caused only a small increase in the diphosphatase activity of minced tissue. Addition of NaCl lowered the Vmax of the diphosphatase reaction.


Simple mathematical deductions in the seroepidemiology of viral infections. I. Herpesvirus group (herpesvirus hominis, varicella-zoster virus, cytomegalovris, Epstein-Barr-Virus). Large samples of nonselected persons collected in South-West Germnay were investigated for the prevalence of serum antibodies to the human Herpesviruses HSV, VZV, CMV and EBV. According to "catalytic models", which compare the infection spread to simple chemical reactions of molecules, a mathematical approximation of the serum surveys was performed. Through a new deduction of the exponential function y = k (l - e-r+) a simple way was found to estimate the annual attack rates in percent of the susceptible, seronegative people. While it was possible to represent the prevalence of serum antibodies to VZV by a continuous curve, a biphasic curve to the antibody prevalence rates in the epidemiology of the other Herpesviruses proved to be more adequate indicating changes in hormonal balance and social behaviour. The use of the epidemiologic parameters k and r for the characterization of the test sensitivity was examined for CMV. By evaluation of the mean antibody titres to CMV, HSV, and EBV throughout different age groups, information about the reactivation of the Herpesvirus diseases could be obtained.


Simple mathematical deductions in the seroepidemiology of viral infections. II. (Para) myxoviruses (measles, mumps, influenza B), rubella, enteroviruses (polio, coxsackie B), adenoviruses, and mycoplasma pneumoniae. Large samples of nonselected persons collected in South-West Germany were investigated for the prevalence of serum antibodies to Poliovirus 1-3 and Coxsackievirus B 1-5 (neutralisation test), to Measles, Rubella, and Mumps (hemagglutination inhibition test), Mumps, Influenza B, Adenovirus, and Mycoplasma pneumoniae (complement-fixation test). According to "catalytic models", which compare the infection spread to simple chemical reactions of molecules as self-limiting procedures, a mathematical approximation of the serum surveys was performed. By the use of only two parameters it became possible to calculate the annual attack rates (without regard of age group arrangement and test sensibility), to construct "true" epidemic curves, and to estimate the persistence of humoral immunity in the population investigated for NT and HIT antibodies.


[Laboratory procedures in adenoviruses. IV. The influence of antiglobulin and complement on neutralization and hemagglutination-inhibition (author's transl)]. The neutralization of adenovirus 3,7, and 19 by rabbit antisera is enhanced by additional application of antirabbit globulin. All sera investigated, obtained early or late after the start of immunization, showed two- to sixfold titer rises. Complement also enhances the neutralization; the titer is raised to a greater extent in early than in late sera. To achieve maximal effects, antiglobulin and complement had to be used in fairly high concentration (ca. 10%). In hemagglutination-inhibition the same set of sera showed a titer increase mediated by anti-rabbit globulin (mostly by the factor 2 to 4), while complement was without influence. The neutralization titer of human sera from patients with adenovirus infections, as opposed to the rabbit antisera, was only slightly increased by addition of anti-human globulin or complement. These reagents had likewise negligible influence on the hemagglutination-inhibition of adenovirus 8 by human sera.


[Fecal flora of man. V. communication: The fluctuation of the fecal flora of the healthy adult (author's transl)]. Seven fecal specimens from each of healthy persons, aged 25 to 42 years, during the period of 2 months were analyzed to obtain the knowledge on the day-to-day variation in the same person and the person-to-person variation. Irrespective of individuals, bacteroidaceae, catenabacteria (eubacteria and strict anaerobic lactobacilli), peptostreptococci (anaerobic gram positive cocci) and bifidobacteria constituted the predominant flora. Spirillaceae, megasphaerae and clostridia (except Cl. perfringens) were also found as predominant flora in specific individuals. In all cases, enterobacteria and streptococci constitute the accompanying flora. In addition, veillonallae and lactobacilli were found as predominant flora in most cases. The remaining flora, consisting of Cl. perfringens, staphylococci and yeast, occurred occasionally in low numbers. With regard to the composition of the fecal flora of each person, a great individual variation was recognized and confirmed statistically. Each person had a characteristic fecal flora. In the same person, the numbers of bacteria of the predominating flora were over a 2 months period relatively constant. On the other hand, a great day-to-day variation in the numbers of bacteria of the accompanying flora as well as the remaining flora was demonstrated.


[Pulmonary injury of mice produced by chronically peroral treatment with different substances (author's transl)]. Authors treated mice chronically peroral with microtoxical doses of paraquate, chromiumsulphate, berylliumsulphate and cadmiumsulphate. It becames evident, that the questioned manner of treatment causes a damage of the lungs. It develops an infiltration in the lungtissue, respectively it comes into being a great pulmonary proliferation of cells. Authors emphasize the fact, that the described pathohistological lesion of the lung develops in that case if the poisoning occurs peroral. If the above mentioned phenomenon comes into being on man similarly pneumophthiseology respectively the hygiene enriches with a new problem, with damages of the lungs produced by chemical substances dosed chronically peroral in very small quantity.


[Detoxification of staphylococcal-enterotoxin B in water (author's transl)]. Staphylococcal enterotoxin B contaminated water has been detoxified with calcium hypochlorite used in the water treatment procedure which is available now in order to get drinking water from surface water in emergency cases. Changes in toxin-activity were measured serologically by agar-gel-immuno-precipitation as well as biologically by the "monkey-feeding test" with young macaca mulatta. With an initial concentration of 50 ppm chlorine in tap water resp. 200 ppm in native surface water concentrations of toxin up to 40 fold average emetic dose of rhesus monkey could be inactivated safely. It was pointed out an obvious conformity between precipitation test and monkey-feeding test. The clinical efficiency of SEB on rhesus monkeys has been investigated and discussed subsequently.


Studies on cytologic characteristics of mammary aspiration smears based on histologic types. Cytologic findings of aspiration smears from 146 histologically confirmed mammary were analyzed in this paper. High cellularity, dispersal of cells in the smears and enlarged nuclei of more than 20 mu in major axis were considered to be useful key points to distinguish carcinoma from benign lesions. Irregular piling, variable nuclear shape, cellular fusion and cytoplasmic inclusions were other common findings in cancer specimens and were scarcely noticed in benign cases. However, in some papillotubular carcinomas the above mentioned characteristics were not so dominant. Scirrhous carcinomas sometimes presented small nuclei in the smears. In some fibroadenomas, the smears were occasionally very cellular and revealed dipersal of cells.


Thyroid function and plasma cyclic AMP response to intravenous glucagon. The glucagon stimulated increase in plasma cyclic AMP has been studied in 17 healthy subjects, in 13 hyperthyroid and in 14 hypothyroid patients. Six hyperthyroid and 2 hypothyroid patients were re-investigated after at least 15 months of treatment. The results show: 1) The glucagon stimulated cyclic AMP response is significantly increased in hyperthyroid patients considered as a group, and is reduced in patients with hypothyroidism. 2) Three hyperthyroid and 4 hypothyroid patients showed a normal response to iv glucagon, indicating that the plasma cyclic AMP response to iv glucagon is not a sensitive test for the evaluation of peripheral thyroid states. This suggests that the effects of thyroid hormones in the liver does not necessarily follow the effects in other tissues. 3) Re-investigation of treated patients showed that the cyclic AMP response can be normalized by treatment, both in hyperthyroidism and hypothyroidism. However, in patients treated for hyperthyroidism a hyper-response to glucagon can continue after blood levels of thyroid hormones are reduced to normal. This suggests an inertia in the loss of the hyper-response to glucagon, once a hyperfunction has been induced. A similar inertia in the loss of glucagon sensitivity in hypothyroidism could explain the large number of normal tests in hypothyroid patients.


Hyaline cytoplasmic inclusions in human hepatoma. A case report. A case of hepatoma in a 79-year old woman is reported. The patient had an enlarged liver and a pathological liver scintigram. Percutaneous liver biopsies were performed both with Menghini-and fine needles. The most prominent feature was the presence of hyaline cytoplasmic PAS-negative inclusions in liver parenchymal cells. There was no nuclear atypia. Electron microscopy disclosed two types of cytoplasmic changes. One consisted of a lamellar ultrastructure and was interpreted as a hyperplasia of smooth-surfaced endoplasmic reticulum. The other change consisted of smooth globular non-membrane limited regions containing amorphous or finely fibrillar material. This was interpreted as corresponding to the hyaline inclusions visible in the light microscope. The presence of PAS-negative hyaline cytoplasmic inclusions may thus be a sign of hepatoma. This may be of relevance for the diagnostic considerations of material obtained by fine needle biopsy.


Herpesvirus infection of the respiratory tract electronmicroscopic observation of the virus in cells obtained from a sputum cytology. A case of herpesvirus infection of the respiratory tract in which the virus was seen ultrastructurally in cells obtained from a routine sputum collected for cytology studies, is presented. The method described would seem feasible for additonal studies at the ultrastructural level of cells obtained during collection of material for routine cytology of the respiratory tract. In the present case the virus particles were easily seen and characterized.


Effect of insulin and adrenaline on cyclic AMP in the diaphragm of normal and diabetic rats. The effects of insulin and adrenaline on cyclic AMP (cAMP) levels in diaphragms of normal, streptozotocin-diabetic and insulin-treated diabetic rats were studied. Adrenaline caused a biphasic rise in cAMP with peak values of cAMP within the first few minutes. Diaphragms of diabetic rats showed an increased responsiveness to adrenaline. Injection of insulin to diabetic rats normalized the rise in cAMP after addition of adrenaline. There was no difference in basal levels of cAMP between diaphragms of normal, diabetic or insulin-treated diabetic rats. Insulin in vitro did not affect basal cAMP-levels or the release of cAMP from the tissue but significantly decreased adrenaline-induced peak levels of cAMP. This effect of insulin was abolished by theophylline. The results of the present study suggest that experimental diabetes is associated with changes of the adenylate cyclase and/or phosphodiesterase enzyme activities in skeletal muscle resulting in an increased responsiveness to adrenaline. Since insulin in vitro depressed the adrenaline-induced elevation of cAMP the increased responsiveness in diaphragms of diabetic rats might be attributed to the specific lack of insulin.


In vitro temporal cAMP and cortisol responses to ACTH by the normal human adrenal gland. The in vitro temporal cAMP and cortisol responses of normal human adrenocortical tissue to ACTH were evaluated. ACTH stimulated rapid increased in cAMP levels and cortisol output, however, significantly increased cAMP levels preceded output only at the higher ACTH doses studied. Cortisol output plateaued with ACTH (10--100 MIU/ml) while cAMP levels continued to increase. The findings indicate that the unitary role of cAMP as the second messenger of ACTH action in the human adrenal cannot be assumed from data derived from lower forms and that the mechanism of action of ACTH in the human may be modulated by additional factors.


Effect of potassium on human and rat liver glucose-6-phosphatase. Potassium chloride concentrations of 100-150 mM were shown to inhibit (20-40%) human and rat liver microsomal glucose-6-phosphatase when the substrate was reduced to physiological concentration. When the enzyme was solubilized by treatment of microsomes with A12O3 the inhibition could be observed at greater substrate concentrations. Since potassium deprivation is associated with hyperglycemia in both animals and man, these observations may have physiological significance.


Responses of serum gonadotrophins to LH-releasing hormone and oestrogens in Japanese women with polycystic ovaries. In order to define the abnormality in gonadotrophin secretion in Japanese women with polycystic ovaries (PCO) who rarely show virilization and markedly enlarged ovaries, basal levels of LH and FSH, and responses of serum gonadotrophins to LH-releasing hormone (LH-RH) or oestrogens were determined by radioimmunoassay. Eleven patients with PCO diagnosed by laparotomy or laparoscopy and 30 normal women in the follicular phase were studied. The mean (+/- SD) basal level of LH was significantly higher in patients with PCO than in normal controls (PCO 28.6 +/- 2.4 vs. normal 10.9 +/- 3.0 mIU/ml), while the mean FSH level in PCO patients was not significantly different from that in the normal controls (9.7 +/- 0.7 vs. 11.4 +/- 2.6 mIU/ml). The mean LH/FSH ratio in PCO patients was significantly higher than that in normal controls (3.2 +/- 0.9 vs. 1.0 +/- 0.3). Exaggerated response of LH to LH-RH was observed in PCO patients, while the FSH response was comparable with the normal controls. Ten out of 11 patients with PCO showed LH release exceeding the basal level after bolus iv injection of 20 mg conjugated oestrogens (Premarin), and virtually the same mean net increase in LH from the basal level was obtained in both PCO patients and normal controls. Since the abnormalities in gonadotrophin secretion in Japanese women with PCO are not different from those reported in patients with PCO in Europe and USA, it seems likely that lower incidence of markedly enlarged ovaries and virilization in Japanese patients may be caused by the difference in ovarian response to gonadotrophin.


Ovarian HCG-binding capacity during the oestrous cycle of the rat. Binding in 125I-labelled HCG to rat ovary varies during the oestrus cycle. The highest amount of binding was found in dioestrous-II rats (7.33 X 10(-15) mol of HCG per mg wet weight), while the lowest binding was observed in oestrous rats (1.67 X 10(-15) mol of HCG per mg wet weight) of circulation LH. It is concluded that the low binding capacity of the rat ovary in the oestrous stage is due to masking of the specific receptors by the excessively high LH hormone secretion in pro-oestrus.


Studies of hormonal regulation of metabolism using isolated hepatocytes. A number of enzymatic methods have been developed to prepare hepatocytes using collagenase and hyaluronidase. However, best cell preparations are obtained by using only low concentrations of collagenase and exposing the liver to the enzyme for a very short period of time. These isolated cells with intact cell membranes and large numbers of microvilli on the cell surface respond to hormones at physiological concentrations suggesting that these microvilli contain hormone receptors. In addition, high glycogen content is essential to maintain the in vivo metabolic characteristics of the hepatocytes suggesting that intracellular glycogen plays an important role in the hormonal regulation of metabolism in hepatocytes. Studies with glucagon and insulin on carbohydrate metabolism show that the molar ratios of these hormones control gluconeogenesis and glycogenolysis. Furthermore, in vitro addition of insulin stimulates glycogen synthesis and activates glycogen synthase. Insulin also stimulates protein synthesis in cells containing high glycogen and maintains more normal parallel strands of polyribosomes. Studies with isolated hepatocytes from diabetic, hypophysectomized and adrenalectomized animals show a reduced glucagon response to glycogenolysis. This lack of glucagon response was not due to reduction in glycogen levels. Other hormones such as somatostatin and parathyroid also give rise to alterations in carbohydrate metabolism in isolated hepatocytes.


Effect of corticotropin, steroidogenic inhibitors and hypophysectomy on the nuclear volumes of fasciculata cells in the rat adrenal cortex. The purpose of this investigation was to measure quantitatively nuclear volumes in outer fasciculata cells in adrenal glands of rats subjected to chronic stimulation of steroidogenesis by ACTH, or to chronic inhibition of functional activity by hypophysectomy or by injection of the steroidogenic inhibitos, U-8113 (p-aminophenyl butanone) and SU-4885 (metopirone). Nuclear volumes, after a recovery period from those effects, were also measured. Chronic daily administrations of ACTH for 7, 14, and 30 days led to a progressive, statistically significant increase in nuclear volumes of fasciculata cells. These changes were found to have been reversed 14 days after discontinuation of hormone treatment. Hypophysectomy resulted in a significant decrease in nuclear volumes. The steroidogenic inhibitors SU-4885 and U-8113 had a biphasic effect on the nuclear volumes with an early decrease and subsequent increase to normal values. Since a decrease in nuclear volumes in the adrenal glands of inhibitor-treated subjects occurred in the presence of cellular hypertrophy and a significant increase in adrenal weights, it is concluded that changes in nuclear volumes can be positively correlated with the secretory activity of the cell rather than with the sizes of cells or glands.


Indirect immunofluorescence detection of human IgM and IgG antibodies against herpes simplex virus type 1 induced cell surface antigens. An indirect immunofluorescence method, based on the use of infected Hela cell coverslip cultures was developed to demonstrate human IgM and IgG class antibodies against herpes simplex virus (HSV) type 1 induced cell surface antigens. A total of 35 specimens from 20 patients have been tested; including patients with a clinical diagnosis of HSV type 1 or type 2 primary infection, patients with recurrent HSV infections, patients without any HSV infections, and patients with varicella-zoster virus (VZV) infections. In each patient with a primary HSV infection both IgM and IgG antibody response was observed, while the patients with recurrent HSV infections showed only IgG antibodies. The direct serological typing of HSV infections was not possible because of cross-reacting antibodies both in the IgG and IgM test. No cross-reactivity was found in this test with HSV and VZV antibodies. The HSV fluorescent IgG and IgM antibody titers were found to parallel the highly sensitive HSV radioimmunoassay antibody titers very closely though at a markedly lower level.


T-typing of group A streptococci from clinical specimens: restriction of the number if implied M types in each T-pattern by tests for opacity factor and nicotinamide adenine dinucleotide glycohydrolase. 117 group A streptococci, consecutively isolated from routine throat swab cultures, were T-typed and further characterized according to their enzyme reactions. The results of typing with two different commercial kits of anti-T sera showed good agreement. 89 per cent of the strains were typable by either of these kits. 68 per cent of the strains were OF-positive and 95 per cent NADse-positive. With T-typing alone, 22 per cent of the strains showed agglutination patterns implying only one M type each. When considered together, the T-typing and enzyme reactions made it possible to implicate one M type only for each of 45 per cent of the strains. In a further 34 per cent of the strains, the possible M type of each strain was restricted to four types. However, these findings were hampered by the fact that the T patterns of a number of more seldom met group A streptococci have not been investigated adequately.


Clinical electromyography: definition, application, and innovation. Clinical EMG techniques, useful in the study of patients with neuromuscular disease, are defined. These include measurement of motor and sensory nerve conduction velocities; analysis of the size, shape, and latency of response of muscle and nerve action potentials after nerve stimulation; estimates of nerve excitability; study of the response to repetitive nerve stimulation as a measure of neuromuscular transmission; and observation of the electrical activity of muscle with needle recording electrodes (monopolar, bipolar, and multielectrodes). From a consideration of the application, limitations, and value of these various techniques, it can be concluded logically that to assure the highest probability of reliable information in regard to an individual patient, the choice of technique(s) and the part(s) to be studied should be intimately tied to a clinical analysis of that patient's problem. EMG as a fishing expedition is rarely fruitful; EMG as an exercise in clinical physiology is often exciting.


Primary hyperparathyroidism presenting as anticonvulsant-induced osteomalacia. A patient presented with the classic features of anticonvulsant-induced osteomalacia. Following discontinuance of diphenylhydantoin therapy and repletion with physiologic quantities of vitamin D, hypercalcemia and persistent biochemical hyperparathyroidism developed, and a parathyroid adenoma was removed. A history of nephrolithiasis and hypercalcemia preceding the institution of drug therapy allowed this patient's underlying parathyroid disease to be defined as primary hyperparathyroidism, which had been obscured by anticonvulsant therapy.


Structural changes in the freeze-fractured sarcolemma of ischemic myocardium. Ultrastructural alterations in the sarcolemma of ischemic myocardium were studied with the freeze-fracture technique. In normal myocardial sarcolemma, the P fracture face contained many intramembranous particles which were randomly distributed, while the E fracture face had few intramembranous particles; no structural abnormalities were seen within the lipid bilayer on either face. Myocardium ischemic for 45 minutes displayed no, or only slight, aggregation of intramembranous particles, but upon reperfusion for 5 to 20 minutes, the particles became significantly aggregated in the P face. Scattered nicks within the lipid bilayer were observed on both fracture faces. Intramembranous particles were similarly aggregated in myocardium ischemic for 2 hours; however, the number of nicks were greatly increased on the P and E fracture faces. These structural alterations, which were undetected in thin sections, are likely to be associated with altered function in the sarcolemma of ischemic myocardium.


Lipid accumulation of hypoxic tissue culture cells. Lipid droplets have long been recognized by light microscopy to accumulate in hypoxic cells both in vivo and in vitro. In the present tissue culture experiments, correlative electron microscopic observations and lipid analyses were performed to determine the nature and significance of lipid accumulation in hypoxia. Strain L mouse fibroblasts were grown in suspension culture, both aerobically and under severe oxygen restriction obtained by gassing cultures daily with an 8% CO(2)-92% nitrogen mixture. After 48 hours, hypoxic cells showed an increase in total lipid/protein ratio of 42% over control cells. Most of this increase was accounted for by an elevation in the level of cellular triglyceride from 12.3 +/- 0.9 mug/mg cell protein in aerobic cultures to 41.9 +/- 0.7 in the hypoxic cultures, an increase of 240%. Levels of cellular free fatty acids (FFA) were 96% higher in the hypoxic cultures. No significant changes in the levels of cellular phospholipid or cholesterol were noted. Electron microscopic examination revealed the accumulation of homogeneous cytoplasmic droplets. The hypoxic changes were reversible upon transferring the cultures to aerobic atmospheres with disappearance of the lipid. These experiments indicate that hypoxic injury initially results in triglyceride and FFA accumulation from an inability to oxidize fatty acids taken up from the media and not from autophagic processes, as described in other types of cell injury associated with the sequestration of membranous residues and intracellular cholesterol and phospholipid accumulation.


Macrophage migratory dysfunction in cancer. A mechanism for subversion of surveillance. There is considerable evidence to suggest that macrophages participate in host resistance to the development and spread of cancer. We have, therefore, studied monocytemacrophage function in humans and animals with neoplasms. Approximately 60% of patients with various types of cancer were found to have abnormal monocyte chemotactic responsiveness in vitro, and abnormal chemotaxis was an indicator of poor prognosis in patients with melanoma. By studying patients before and after surgery, it was found that abnormal chemotactic responses normalized within weeks after removal of malignant tumors, indicating that a neoplasm itself might affect the host's monocyte chemotactic responsiveness. Subsequent studies using transplantable neoplasms in mice substantiated this hypothesis in that macrophage accumulation in vivo as well as macrophage chemotactic responsiveness in vitro was depressed in animals during the early phases of tumor growth. This depression of macrophage function could be attributed to a low-molecular-weight factor contained in murine neoplasms, which when given to normal mice was extremely potent in depressing peritoneal macrophage accumulation and chemotaxis but, paradoxically, enhanced phagocytosis. The serum of tumor-bearing mice also contained potent inhibitory activity for macrophage accumulation. In contrast to the effects on macrophages, granulocyte accumulation in vivo and chemotaxis in vitro was not depressed by the presence of a neoplasm or the administration of the factor from neoplasms. By releasing factors which depress macrophage migratory function, neoplasms may protect themselves from immunologically mediated host destruction during the early phases of tumor growth.